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WO2018154466A1 - Dihydroquinolizinones as antivirals - Google Patents

Dihydroquinolizinones as antivirals Download PDF

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Publication number
WO2018154466A1
WO2018154466A1 PCT/IB2018/051080 IB2018051080W WO2018154466A1 WO 2018154466 A1 WO2018154466 A1 WO 2018154466A1 IB 2018051080 W IB2018051080 W IB 2018051080W WO 2018154466 A1 WO2018154466 A1 WO 2018154466A1
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alkyl
substituted
unsubstituted
halo
alkoxy
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French (fr)
Inventor
John G. Catalano
Hamilton D. Dickson
Wieslaw Mieczyslaw Kazmierski
Martin R. LEIVERS
John Gordon WEATHERHEAD
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GlaxoSmithKline Intellectual Property Development Ltd
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GlaxoSmithKline Intellectual Property Development Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
    • C07D491/20—Spiro-condensed systems

Definitions

  • the present invention relates to compounds useful for treatment of HBV in animals, and more particularly for treatment of HBV in humans.
  • Hepatitis B is a viral disease transmitted parenterally by contaminated material such as blood and blood products, contaminated needles, sexually and vertically from infected or carrier mothers to their offspring. In those areas of the world where the disease is common vertical transmission at an early age results in a high proportion of infected individuals becoming chronic carriers of hepatitis B. It is estimated by the World Health Organization that more than 2 billion people have been infected worldwide, with about 4 million acute cases per year, 1 million deaths per year, and 350-400 million chronic carriers. Approximately 25% of carriers die from chronic hepatitis, cirrhosis, or liver cancer and nearly 75% of chronic carriers are Asian. Hepatitis B virus (HBV) is the second most significant carcinogen behind tobacco, causing from 60% to 80% of all primary liver cancer. HBV is 100 times more contagious than HIV.
  • HBV Hepatitis B virus
  • HBV is transmitted through percutaneous or parenteral contact with infected blood, body fluids, and by sexual intercourse. HBV is able to remain on any surface it comes into contact with for about a week, e.g. table-tops, razor blades, blood stains, without losing infectivity.
  • HBV cannot cross the skin or the mucous membrane barrier. Some break in this barrier, which can be minimal and insignificant, is required for transmission.
  • HBV is a small enveloped DNA virus belonging to the hepadnavirus family.
  • the virus replicates through an RNA intermediate form by reverse transcription, which in practice relates them to retroviruses, like HIV. Although replication takes place in the liver, the virus spreads to the blood where viral proteins and antibodies against them are found in infected people. HBV is many times more infectious than HIV due to the greater concentrations of HBV virus found in the bloodstream at any given time.
  • HBV infection results in the production of two different particles: 1 ) the HBV virus itself (or Dane particle) which includes a viral capsid assembled from the HBV core antigen protein (HBcAg) and is covered by the hepatitis B surface antigen (HBsAg) and is capable of reinfecting cells and 2) subviral particles (or SVPs) which are high density lipoprotein-like particles comprised of lipids, cholesterol, cholesterol esters and the small and medium forms of the hepatitis B surface antigen (HBsAg) which are non-infectious.
  • SVPs subviral particles
  • HBV infected cells also secrete a soluble proteolytic product of the pre-core protein called the HBV e-antigen (HBeAg).
  • HBV e-antigen HBV e-antigen
  • HDV hepatitis D virus
  • HBsAg hepatitis D virus
  • HDV infection can only occur in subjects with concomitant HBV infection. While the incidence of HDV co-infection in asymptomatic HBV carriers and chronic HBV- related liver disease is low in countries with a low incidence of HBV infection, it is a significant complication in HBV-infected subjects in countries with a high incidence of HBV infection and can increase the rate of progression of liver disease to fulminant hepatitis. As such, the clear unmet medical need in HBV infection is even more pressing in HBV/HDV co-infected subjects.
  • HBV polymerase inhibitors are effective in reducing viral production, but have little to no effect in rapidly reducing HBsAg blood levels or can slowly reduce HBsAg with long term treatment in a limited number of patients (as is the case with tenofovir disoproxil fumarate).
  • Interferon based immunotherapy can achieve a reduction of both viral production and early removal of HBsAg from the blood but only in a small percentage of treated subjects.
  • HBsAg The generally accepted role of HBsAg in the blood is to sequester anti-HBsAg antibodies and allow infectious viral particles to escape immune detection which is likely one of the reasons why HBV infection remains a chronic condition.
  • HBsAg, HBeAg and HBcAg all have immuno-inhibitory properties as discussed below and the persistence of these viral proteins in the blood of patients following the administration of any of the currently available treatments for HBV as described above is likely having a significant impact in preventing patients from achieving immunological control of their HBV infection.
  • HBsAg comprises the overwhelming majority of HBV protein in the circulation of HBV infected subjects. Additionally, while the removal (via
  • Hepatitis B viral infections in conjunction with Hepatitis D viral infections, are a continuing medical problem because, like any rapidly-replicating infectious agent, there are continuing mutations that help some sub-populations of HBV become resistant to current treatment regimens.
  • HBV Hepatitis D virus
  • HDV Hepatitis D virus
  • nucleoside and nucleotide therapies entecavir and tenofovir are successful at reducing viral load, but the rates of HBeAg seroconversion and HBsAg loss are even lower than those obtained using IFNa therapy.
  • Other similar therapies including lamivudine (3TC), telbivudine (LdT), and adefovir are also used, but for nucleoside/nucleotide therapies in general, the emergence of resistance limits therapeutic efficacy.
  • An embodiment of the present invention features a compound of Formula I:
  • R 1 and R 4 are independently selected from hydrogen; halogen; hydroxy; alkoxy; amino; cyano; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl;
  • R 2 and R 3 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR 20 R 21 ; COR 19 ; COOR 19 ; CONR 20 R 21 ; pyrrolidinyl; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; Ci- 6 alkylCOOR 19 ; -BOR 19 OR 19' ; unsubstituted Ci- 6 alkoxy or halo- or alkyl-substituted Ci- 6 alkyoxy; unsubstituted C 3 . 7 cycloalkyl or halo- or alkyl-substituted C 3 .
  • R 7 and R 8 are selected from independently hydrogen; halogen; cyano; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- or alkyl-substituted Ci_6alkoxy; unsubstituted C 3 -8cycloalkyl or halo- or alkyl-substituted C 3 - 8 cycloalkyl; unsubstituted C x H 2 x-phenyl or halo- or alkyl-substituted -C x H 2 x-phenyl; and unsubstituted -0-C x H 2x -phenyl or halo- or alkyl-substituted -0-C x H 2x -phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R 7 and R 8 together form
  • R 9 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- or alkyl- substituted Ci- 6 alkoxy; unsubstituted C 3 - 8 cycloalkyl or halo- or alkyl-substituted C 3 .
  • ecycloalkyl unsubstituted or halo- or alkyl-substituted -C x H 2x -phenyl; and unsubstituted or halo- or alkyl substituted -0-C x H 2x -phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
  • R 10 is selected from COOR 19 and CONR 20 R 21 6;
  • R 11 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci- 6 alkyoxy; unsubstituted or halo- or alkyl-substituted C 3 . 7 cycloalkyl;
  • R 12 is selected from hydrogen; unsubstituted Ci_ 6 alkyl or Ci_ 6 alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, heterocycloalkyi, heteroaryl, C 3 . 8 cycloalkyl and C 2 . 6 alkenyl; Ci- 6 alkylCi- 6 alkoxy; Ci_ 6 alkylCi- 6 alkoxyCi- 6 alkoxy; Ci.
  • heteroaryl is a monocyclic heteroaryl comprising O and/or ; and Ci- 6 alkylheterocycloalkyl, wherein the heterocycloalkyi is a monocyclic heterocycloalkyi comprising O and/or N;
  • R 13 , R 13' , R 14 and R 14' are independently selected from hydrogen,; halogen; amino; aminoalkyl; cyano; Ci_ 6 alkyl; Ci- 6 alkoxy; carbonyl; carboxamide; and amide; or R 13 and R 13 ' or R 14 and R 14' together form a 3- to 8-membered cycloalkyl; or R 14 and R 14' together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_ 6 alkyl, C 3 - 8 cycloalkyl, C 2 - 6 alkenyl or Ci- 6 alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR 22 , S, SR 22 or SR 22 R 22 ';
  • R 17 , R 17 ', R 18 and R 18' are independently selected from hydrogen; halogen; amino; cyano; Ci_ 6 alkyl; Ci- 6 alkoxy; carbonyl; carboxamide; and amide; or R 17 and R 18 or R 17' and R 18' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci- 6 alkyl, C 3 - 8 cycloalkyl, C 2 - 6 alkenyl or Ci- 6 alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR 22 , S, SR 22 or SR 22 R 22' ;
  • R 19 , R 19' and R 19" are independently selected from hydrogen; Ci_ 6 alkyl; C 3 . ecycloalkyl; C 2 . 6 alkenyl; phenyl; Ci- 6 alkylimidizole; Ci- 6 alkoxy; Ci- 6 alkyltriazole; Ci_
  • R 20 and R 21 are independently selected from hydrogen; Ci_ 6 alkyl; C 3 .
  • Ci_ ecycloalkyl; C 2 . 6 alkenyl; phenyl; Ci- 6 alkylimidizole; Ci- 6 alkoxy; Ci- 6 alkyltriazole; Ci_
  • R 20 and R 21 together with the nitrogen to which they are attached form unsubstituted pyrrolidinyl, unsubstituted piperidinyl, or unsubstituted morpholinyl; or R 20 and R 21 together with the nitrogen to which they are attached form carboxyl-substituted pyrrolidinyl, carboxyl-substituted piperidinyl or carboxyl-substituted morpholinyl;
  • R 22 and R 22' are independently selected from hydrogen; oxygen; Ci_ 6 alkyl; Ci- 6 haloalkyl; C 3 . 8 cycloalkyl; Ci- 6 alkylC 3 - 8 cycloalkyl; C 2 .
  • R 23 is selected from hydrogen, Ci_ 6 alkyl; C 3 - 8 cycloalkyl; C 2 - 6 alkenyl; phenyl; Ci- 6 alkylimidizole; Ci- 6 alkoxy; Ci- 6 alkyltriazole; Ci- 6 alkyltetrazole; Ci- 6 alkylthiazole; Ci- 6 alkyloxazole; Ci_ ealkyldioxazole; Ci- 6 alkyloxazolidone; -COR 19 ; -COOR 19' ; -CSOR 19 " ; and -CONR 20 R 21 ; and [0028]
  • R 23 is selected from hydrogen, Ci_ 6 alkyl; C 3 - 8 cycloalkyl; C 2 - 6 alkenyl; phenyl; Ci- 6 alkylimidizole; Ci- 6 alkoxy; Ci- 6 alkyltriazole; Ci- 6 alkyltetrazole; Ci- 6 alkylthiazole
  • One embodiment of the invention provides a compound of Formula I
  • R 1 and R 4 are independently selected from hydrogen; halogen; hydroxy; alkoxy; amino; cyano; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl;
  • R 2 and R 3 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR 20 R 21 ; COR 19 ; COOR 19 ; CONR 20 R 21 ; pyrrolidinyl; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; Ci- 6 alkylCOOR 19 ; -BOR 19 OR 19' ; unsubstituted Ci- 6 alkoxy or halo- or alkyl-substituted Ci- 6 alkyoxy; unsubstituted C 3 . 7 cycloalkyl or halo- or alkyl-substituted C 3 . 7 cycloalkyl; C 3 . 7 cycloalkenyl or halo- or alkyl-substituted C 3 .
  • R 5 is selected from hydrogen and unsubstituted Ci_ 6 alkyl or halo- or alkyl- substituted Ci_ 6 alkyl;
  • R 6 and R 7 together form a 3- to 8-membere cycloalkyl ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR 23 , R 15 , R 15' , R 16 and/or R 16' ; or R 6 and R 7 together form a 3- to 8- membered heterocycloalkyi ring comprising one heteroatom or two or more heteroatoms, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R 15 , R 15' , R 16 and/or R 16' , wherein the one heteroatom in the heterocycloalkyi ring is NR 20 and the two or more heteroatoms are selected from N, NR 22 , O, S, SR 22 and SR 22 R 22' ;
  • R 8 is selected from hydrogen and unsubstituted Ci_ 6 alkyl or halo- or alkyl- substituted Ci_ 6 alkyl;
  • R 9 is selected from hydrogen, hydroxy, halogen, cyano, unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl, unsubstituted Ci- 6 alkoxy or halo- or alkyl- substituted Ci- 6 alkoxy, unsubstituted C 3 - 8 cycloalkyl or halo- or alkyl-substituted C 3 .
  • ecycloalkyl unsubstituted -C x H 2 x-phenyl or halo- or alkyl-substituted -C x H 2x -phenyl an unsubstituted -0-C x H 2x -phenyl or halo- or alkyl-substituted -0-C x H 2 x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
  • R 10 is selected from COOR 19 and CONR 20 R 21 ;
  • R 11 is selected from hydrogen, hydroxy, halogen, cyano, unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl, unsubstituted Ci- 6 alkoxy or halo- or alkyl- substituted Ci- 6 alkyoxy; unsubstituted C 3 - 7 cycloalkyl or halo- or alkyl substituted C 3 .
  • ycycloalkyl unsubstituted -C x H 2x -phenyl or halo- or alkyl-substituted -C x H 2x -phenyl; and unsubstituted -0-C x H 2x -phenyl or halo- or alkyl-substituted -0-C x H 2x -phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
  • R 12 is hydrogen; unsubstituted Ci_ 6 alkyl or Ci_ 6 alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, heterocycloalkyi, heteroaryl, C 3 . 8 cycloalkyl and C 2 . 6 alkenyl; Ci- 6 alkylCi- 6 alkoxy; Ci- 6 alkylCi- 6 alkoxyCi- 6 alkoxy; Ci- 8 alkylamino; Ci- 8 alkylaminocarbonylCi- 6 alkyl; Ci- 6 alkylaminosulfonylCi- 6 alkyl; Ci_
  • R 15 , R 15' , R 16 and R 16' are independently hydrogen, halogen, amino, cyano, Ci- 6 alkyl, or Ci- 6 alkoxy, wherein any or R 15 , R 15' , R 16 or R
  • R 19 , R 19' and R 19" are independently hydrogen, Ci_ 6 alkyl, C 3 - 8 cycloalkyl, C 2 . 6 alkenyl, phenyl, Ci- 6 alkylimidizole, Ci- 6 alkoxy, Ci- 6 alkyltriazole, Ci- 6 alkyltetrazole, Ci_
  • R 20 and R 21 are independently hydrogen, Ci_6alkyl, C 3 -8cycloalkyl, C 2 .
  • R 22 and R 22' are independently selected from hydrogen, oxygen, Ci_ 6 alkyl, Ci- 6 haloalkyl, C 3 - 8 cycloalkyl, Ci- 6 alkylC 3 - 8 cycloalkyl, C 2 .
  • R 23 is hydrogen, Ci_ 6 alkyl, C 3 . 8 cycloalkyl, C 2 . 6 alkenyl, phenyl, Ci_
  • 6alkyloxazole Ci. 6 alkyldioxazole; Ci. 6 alkyloxazolidone; aryl, heteroaryl or benzyl, or a pharmaceutically acceptable salt thereof.
  • One embodiment of the invention provides a compound of Formula I
  • R 1 , and R 4 are independently selected from hydrogen; halogen; hydroxy; alkoxy; amino; cyano; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl;
  • R 2 and R 3 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR 20 R 21 ; COR 19 ; COOR 19 ; CONR 20 R 21 ; pyrrolidinyl; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; Ci- 6 alkylCOOR 19 '; -BOR 19 OR 19' ; unsubstituted Ci- 6 alkoxy or halo- or alkyl-substituted Ci- 6 alkyoxy; C 3 - 7 cycloalkyl or halo- or alkyl-substituted C 3 - 7 cycloalkyl; unsubstituted C 3 .
  • R 5 and R 6 are independently selected from hydrogen; halogen; cyano; unsubstituted Ci_ 6 alkyl or halo-or alkyl-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- or alkyl-substituted Ci_6alkoxy; unsubstituted C 3 -ecycloalkyl or halo- or alkyl-substituted C 3 .
  • ecycloalkyl unsubstituted -C x H 2x -phenyl or halo- or alkyl-substituted -C x H 2x -phenyl; and unsubstituted -0-C x H 2x -phenyl or halo- or alkyl-substituted -0-C x H 2x -phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R 5 and R 6 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR 23 , R 13 , R 13' , R 14 and/or R 14' , or R 5 and R 6 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R 13 ,
  • R 7 and R 8 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR 23 , R 17 , R 17' , R 18 and/or R 18' , or R 7 and R 8 together form a 3- to 8- membered
  • heterocycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R 17 , R 18 ', R 17 and/or R 18' , wherein a heteroatom in the heterocycloalkyi ring is O, N, NR 22 , S, SR 22 or SR 22 R 22' ;
  • R 9 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6 alkyl or halo-or alkyl-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3-ecycloalkyl or halo- or alkyl-substituted C3- ecycloalkyl; unsubstituted -C x H 2 x-phenyl or halo- or alkyl-substituted -C x H 2 x-phenyl; and unsubstituted -0-C x H 2x -phenyl or halo- or alkyl-substituted -0-C x H 2x -phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
  • R 10 is selected from COOR 19 and CONR 20 R 21 ;
  • R 11 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- or alkyl- substituted Ci- 6 alkyoxy; unsubstituted or halo- or alkyl-substituted C 3 -7cycloalkyl;
  • R 12 is selected from hydrogen; unsubstituted Ci_ 6 alkyl or Ci_ 6 alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, C 3 - ecycloalkyl and C 2 .
  • R 13 , R 13' , R 14 and R 14' are independently selected from hydrogen; halogen; amino; aminoalkyl; cyano; Ci_ 6 alkyl; Ci- 6 alkoxy; carbonyl; carboxamide; and amide; or R 13 and R 14 or R 13 ' and R 14' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_ 6 alkyl, C 3 - 8 cycloalkyl, C 2 - 6 alkenyl or Ci- 6 alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR 22 , S, SR 22 or SR 22 R 22' ;
  • R 17 , R 17 ', R 18 and R 18' are independently selected from hydrogen; halogen; amino; cyano; Ci_ 6 alkyl; and Ci- 6 alkoxy; or R 17 and R 18 or R 17' and R 18' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_ 6 alkyl, C 3 - 8 cycloalkyl, C 2 . 6 alkenyl or Ci- 6 alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR 22 , S, SR 22 or SR 22 R 22' ;
  • R 19 , R 19' and R 19" are independently selected from hydrogen; Ci-ealkyl; C 3 . ecycloalkyl; C 2 - 6 alkenyl; phenyl; Ci- 6 alkylimidizole; Ci- 6 alkoxy; Ci- 6 alkyltriazole; Ci_
  • R 20 and R 21 are independently selected from hydrogen; Ci_ 6 alkyl; C 3 . ecycloalkyl; C 2 -6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci- 6 alkyltetrazole; Ci- 6 alkylthiazole; Ci- 6 alkyloxazole; Ci- 6 alkyldioxazole; and Ci_
  • R 20 and R 21 together with the nitrogen to which they are attached form unsubstituted pyrrolidinyl, unsubstituted piperidinyl, or unsubstituted morpholinyl; or form carboxyl-substituted pyrrolidinyl, carboxyl-substituted piperidinyl or carboxyl-substituted morpholinyl; and
  • R 22 and R 22' are independently selected from hydrogen; oxygen; Ci_ 6 alkyl; Ci- 6 haloalkyl, C 3 - 8 cycloalkyl; Ci- 6 alkylC 3 - 8 cycloalkyl; C 2 - 6 alkenyl; phenyl; Ci- 6 alkylimidizole; Ci- 6 alkoxy; Ci- 6 alkyltriazole; Ci- 6 alkyltetrazole; Ci- 6 alkylthiazole; Ci- 6 alkyloxazole; Ci_ ealkyldioxazole; Ci- 6 alkyloxazolidone; -COR 19 ; -COOR 19' ; -CSOR 19 " ; and -CONR 20 R 21 ; and
  • R 23 is selected from hydrogen; Ci_ 6 alkyl; C 3 . 8 cycloalkyl; C 2 - 6 alkenyl; phenyl; Ci- 6 alkylimidizole; Ci- 6 alkoxy; Ci- 6 alkyltriazole; Ci- 6 alkyltetrazole; Ci- 6 alkylthiazole; Ci_
  • Another embodiment provides a compound of Formula IA or Formula IB:
  • C* is a carbon atom stereocenter which has a configuration which is (R) or (S);
  • R 1 , R 2 , R 3 and R 4 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR 20 R 21 COR 19 ; COOR 19 ; CONR 20 R 21 ; pyrrolidinyl; unsubstituted Ci- 6 alkyl or halo- or alkyl substituted Ci_ 6 alkyl; Ci- 6 alkylCOOR 19 ; -BOR 19 OR 19' ; unsubstituted Ci- 6 alkoxy or halo-or alkyl-substituted Ci- 6 alkyoxy; unsubstituted C 3 - 7 cycloalkyl or halo-or alkyl-substituted C 3 - 7 cycloalkyl; unsubstituted C 3 - 7 cycloalkenyl or halo- or alkyl-substituted C 3 - 7 cycloalkenyl; unsubstituted or halo- or cyano
  • R 5 and R 6 are independently selected from halogen; cyano;
  • R 7 and R 8 are independently hydrogen; halogen; cyano; unsubstituted Ci_ 6alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3-ecycloalkyl or halo- or alkyl-substituted C3- ecycloalkyl; unsubstituted or halo- or alkyl-substituted-C x H 2 x-phenyl; unsubstituted or halo- or alkyl-substituted -0-C x H 2x -phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R 7 and R 8 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, al
  • R 5 and R 8 are independently selected from hydrogen, unsubstituted Ci_ 6 alkyl, and halo- or alkyl-substituted Ci_ 6 alkyl;
  • R 6 and R 7 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR 23 , R 15 , R 15' , R 16 and/or R 16' ; or R 6 and R 7 together form a 3- to 8- membered heterocycloalkyi ring comprising one heteroatom or two or more heteroatoms, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R 15 , R 15' , R 16 and/or R 16' , wherein the one heteroatom in the heterocycloalkyi ring is NR 20 and the two or more heteroatoms are selected from N, NR 22 , O, S, SR 22 and SR 22 R 22' ; or
  • R 5 and R 6 are independently selected from hydrogen; halogen; cyano; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- or alkyl-substituted Ci- 6 alkoxy; unsubstituted C 3 - 8 cycloalkyl or halo- or alkyl- substituted C 3 - 8 cycloalkyl; unsubstituted or halo- or alkyl-substituted -C x H 2x -phenyl;
  • R 5 and R 6 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR 23 , R 17 , R 17' , R 18 and/or R 18' , or R 5 and R 6 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R 13 , R 13 ', R 14 and/or R 14' , wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR 22 , S, SR 22 or SR 22 R 22' ; and
  • R 7 and R 8 are independently selected from halogen; cyano; unsubstituted Ci- 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3-ecycloalkyl or halo- or alkyl-substituted C3- ecycloalkyl; unsubstituted or halo- or alkyl-substituted -C x H 2x -phenyl; unsubstituted or halo- or alkyl-substituted -0-C x H 2x -phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; and heteroaryl; or R 7 and R 8 together form a 3- to 8-membered cycloalkyi ring optionally substituted with ox
  • R 9 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- or alkyl- substituted Ci- 6 alkoxy; unsubstituted C 3 - 8 cycloalkyl or halo- or alkyl-substituted C 3 .
  • ecycloalkyl unsubstituted or halo- or alkyl-substituted -C x H 2x -phenyl; and unsubstituted or halo- or alkyl-substituted-0-C x H 2) ⁇ phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
  • R 10 is selected from COOR 19 and CONR 20 R 21 ;
  • R 11 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- or alkyl- substituted Ci- 6 alkyoxy; unsubstituted or halo- or alkyl-substituted C 3 - 7 cycloalkyl;
  • R 12 is selected from hydrogen; unsubstituted Ci_ 6 alkyl or Ci_ 6 alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, heterocycloalkyi, heteroaryl, C 3 - 8 cycloalkyl and C 2 - 6 alkenyl; Ci- 6 alkylCi- 6 alkoxy; Ci_ 6 alkylCi- 6 alkoxyCi- 6 alkoxy; Ci.
  • heteroaryl is an O-containing or N-containing monocyclic heteroaryl
  • R 13 , R 13' , R 14 and R 14' are independently selected from hydrogen; halogen; amino; aminoalkyl; cyano; Ci_6alkyl; Ci-6alkoxy; carbonyl; carboxamide; and amide; or R 13 and R 13 ' or R 14 and R 14' together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C 3 -ecycloalkyl, C 2 -6alkenyl or Ci_6alkoxy, wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR 22 , S, SR 22 or SR 22 R 22' ; [0074] R 15 , R 15' , R 16 and R 16' are independently hydrogen, halogen, amino, cyano, Ci- 6 alkyl, or Ci-
  • R 17 , R 17 ', R 18 and R 18' are independently selected from hydrogen; halogen; amino; cyano; Ci_ 6 alkyl; and Ci- 6 alkoxy; or R 17 and R 18 or R 17' and R 18' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_ 6 alkyl, C 3 - 8 cycloalkyl, C 2 .
  • 6alkenyl or Ci_6alkoxy wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR 22 , S, SR 22 or SR 22 R 22' ;
  • R 19 , R 19' and R 19" are independently selected from hydrogen; Ci- 6 alkyl; C 3 . ecycloalkyl; C 2 - 6 alkenyl; phenyl; Ci- 6 alkylimidizole; Ci- 6 alkoxy; Ci- 6 alkyltriazole; Ci_
  • R 20 and R 21 are independently selected from hydrogen; Ci_ 6 alkyl; C 3 .
  • Ci_ ecycloalkyl; C 2 . 6 alkenyl; phenyl; Ci- 6 alkylimidizole; Ci- 6 alkoxy; Ci- 6 alkyltriazole; Ci_
  • R 20 and R 21 together with the nitrogen to which they are attached form unsubstituted pyrrolidinyl, unsubstituted piperidinyl, or unsubstituted morpholinyl; or form carboxyl-substituted pyrrolidinyl, carboxyl-substituted piperidinyl or carboxyl-substituted morpholinyl; and
  • R 22 and R 22' are independently selected from hydrogen; oxygen; Ci_ 6 alkyl; Ci- 6 haloalkyl; C 3 . 8 cycloalkyl; Ci- 6 alkylC 3 - 8 cycloalkyl; C 2 .
  • R 23 is selected from hydrogen; Ci_ 6 alkyl; C 3 . 8 cycloalkyl; C 2 . 6 alkenyl; phenyl; Ci- 6 alkylimidizole; Ci- 6 alkoxy; Ci- 6 alkyltriazole; Ci. 6 alkyltetrazole; Ci. 6 alkylthiazole; Ci_
  • a particular embodiment provides a compound selected from the group consisting of: 9'-(Benzyloxy)-10'-bromo-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
  • a particular embodiment provides a compound from Table 1 B, or a pharmaceutically acceptable salt thereof.
  • the present invention is directed to treatment the treatment of a hepatitis B infection or hepatitis B/hepatitis D co-infection in a human subject. Therefore, in certain embodiments, the present invention provides a method for the treatment of a hepatitis B infection or hepatitis B/hepatitis D co-infection, the method comprising administering to a subject in need of such treatment a compound of Formula I, IA or IB or a compound selected from Table 1A and Table 1 B as described herein.
  • the present invention provides a method for the treatment of a hepatitis B infection or hepatitis B/hepatitis D co-infection, the method comprising administering to a subject in need of such treatment a first pharmaceutically acceptable agent comprising a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1A and Table 1 B as described as described herein, in combination with a second pharmaceutically acceptable agent that stimulates immune function and a third pharmaceutically acceptable agent comprising an antiviral compound.
  • the administration of a compound of Formula I, IA or IB or a compound selected from Table 1 A and Table 1 B as described as described herein inhibits the release of hepatitis B surface antigen (HBsAg), HB core antigen protein (HBcAg), and/or hepatitis B pre-core protein known as the HBV e-antigen antigen (HBeAg) from infected hepatocytes.
  • HBsAg hepatitis B surface antigen
  • HBcAg HB core antigen protein
  • HBV eAg hepatitis B pre-core protein known as the HBV e-antigen antigen
  • Embodiments of the present invention features compounds that inhibit levels of HBe and/or HBs antigens in a subject infected with hepatitis B virus, and therefore are useful for treating human hepatitis B virus infections, and disease and symptoms associated with such virus infections.
  • Table 1 A and Table 1 B show a listing of compounds and structures, with ELISA data showing EC 5 o value ranges measured against the HBs antigen in a Hep AD 38 cell line (uM), for compounds from Table 1 A described herein.
  • alkyl refers to a monovalent saturated aliphatic hydrocarbyl group having from 1 to 14 carbon atoms and, in some embodiments, from 1 to 6 carbon atoms
  • alkyl includes, by way of example, linear and branched hydrocarbyl groups such as methyl (CH 3 -), ethyl (CH 3 CH 2 -), n-propyl
  • CH3CH2CH2- isopropyl ((CH 3 ) 2 CH-), n-butyl (CH3CH2CH2CH2-), isobutyl ((CH 3 ) 2 CHCH 2 -), sec-butyl ((CH 3 )(CH 3 CH 2 )CH-), /-butyl ((CH 3 ) 3 C-), ⁇ -pentyl (CH 3 CH2CH 2 CH 2 CH2-), and neopentyl ((CH 3 ) 3 CCH 2 -).
  • AlkyI groups may also be substituted, for example, with one or more alkyl, cycloalkyl, heterocycloalkyl, alkoxy, carboxy, amido, keto, amino, cyano, aryl, heteroaryl, halo or haloalkyl substituents.
  • Alkoxy refers to the group -O-alkyl wherein alkyl is defined herein. Alkoxy includes, by way of example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, f-butoxy, sec-butoxy, n-pentoxy, morpholinylpropoxy, piperidinylethoxy.
  • Amino refers to the group -NR a R b where R a and R b are independently selected from hydrogen, alkyl, alkenyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, heterocyclic, and wherein R a and R b are optionally joined together with the nitrogen bound thereto to form a heterocyclic group.
  • R a is hydrogen and R b is alkyl
  • the amino group is sometimes referred to herein as alkylamino.
  • R a and R b are alkyl
  • dialkylamino When referring to a monosubstituted amino, it is meant that either R a or R b is hydrogen but not both.
  • a disubstituted amino it is meant that neither R a nor R b are hydrogen.
  • Aryl refers to an aromatic group of from 5 to 14 carbon atoms and no ring heteroatoms and having a single ring (e.g. , phenyl) or multiple condensed (fused) rings (e.g. , naphthyl or anthryl).
  • a single ring e.g. , phenyl
  • multiple condensed (fused) rings e.g. , naphthyl or anthryl.
  • the term “Aryl” or “Ar” applies when the point of attachment is at an aromatic carbon atom (e.g. , 5,6,7,8 tetrahydronaphthalene-2-yl is an aryl group as its point of attachment is at the 2- position of the aromatic phenyl ring).
  • Cycloalkyl refers to a saturated or partially saturated cyclic group of from 3 to 14 carbon atoms and no ring heteroatoms and having a single ring or multiple rings including fused, bridged, and spiro ring systems.
  • cycloalkyl applies when the point of attachment is at a non-aromatic carbon atom (e.g. 5,6,7,8,- tetrahydronaphthalene-5-yl).
  • Cycloalkyl includes cycloalkenyl groups, such as cyclohexenyl.
  • cycloalkyl groups include, for instance, adamantyl, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclooctyl, cyclopentenyl, and cyclohexenyl.
  • cycloalkyl groups that include multiple bicycloalkyl ring systems are bicyclohexyl, bicyclopentyl, bicyclooctyl, and the like.
  • Cycloalkyl groups may also be substituted, for example, with one or more alkyl, cycloalkyl, heterocycloalkyl, alkoxy, carboxy, amido, keto, amino, cyano, aryl, heteroaryl, halo or haloalkyl substituents.
  • Halo or "halogen” refers to fluoro, chloro, bromo, and iodo.
  • Haloalkyl refers to substitution of alkyl groups with 1 to 9 (e.g. when the alkyl group has 3 carbon atoms, such as a t-butyl group fully-substituted with halogen) or in some embodiments 1 to 3 halo groups (e.g. trifluoromethyl).
  • Heteroaryl refers to an aromatic group of from 1 to 14 carbon atoms and 1 to 6 heteroatoms selected from oxygen, nitrogen, sulfur, phosphorus, silicon and boron, and includes single ring (e.g. imidazolyl) and multiple ring systems (e.g. benzimidazol-2-yl and benzimidazol-6-yl).
  • single ring e.g. imidazolyl
  • multiple ring systems e.g. benzimidazol-2-yl and benzimidazol-6-yl
  • the term “heteroaryl” applies if there is at least one ring heteroatom and the point of attachment is at an atom of an aromatic ring (e.g.
  • the nitrogen and/or the sulfur ring atom(s) of the heteroaryl group are optionally oxidized to provide for the N-oxide (N ⁇ 0), sulfinyl, or sulfonyl moieties.
  • heteroaryl includes, but is not limited to, pyridyl, furanyl, thienyl, thiazolyl, isothiazolyl, triazolyl, imidazolyl, imidazolinyl, isoxazolyl, pyrrolyl, pyrazolyl, pyridazinyl, pyrimidinyl, purinyl, phthalazyl, naphthyl, naphthylpryidyl, oxazolyl, quinolyl, benzofuranyl,
  • quinazolinonyl benzimidazolyl, benzisoxazolyl, benzothienyl, benzopyridazinyl, pteridinyl, carbazolyl, carbolinyl, phenanthridinyl, acridinyl, phenanthrolinyl, phenazinyl, phenoxazinyl, phenothiazinyl, and phthalimidyl.
  • Heterocyclic or “heterocycle” or “heterocycloalkyl” or “heterocyclyl” refers to a saturated or partially saturated cyclic group having from 1 to 14 carbon atoms and from 1 to 6 heteroatoms selected from nitrogen, sulfur, phosphorus or oxygen and includes single ring and multiple ring systems including fused, bridged, and spiro ring systems. For multiple ring systems having aromatic and/or non-aromatic rings, the terms “heterocyclic",
  • heterocycle when there is at least one ring heteroatom and the point of attachment is at an atom of a non-aromatic ring (e.g. 1 ,2,3,4- tetrahydroquinoline-3-yl, 5,6,7,8-tetrahydroquinoline-6-yl, and decahydroquinolin-6-yl).
  • the nitrogen, phosphorus and/or sulfur atom(s) of the heterocyclic group are optionally oxidized to provide for the N-oxide, phosphinane oxide, sulfinyl, sulfonyl moieties.
  • heterocyclyl includes, but is not limited to, tetrahydropyranyl, piperidinyl, piperazinyl, 3-pyrrolidinyl, 2-pyrrolidon-1 -yl, morpholinyl, and pyrrolidinyl.
  • a prefix indicating the number of carbon atoms e.g., C 3 -Ci 0 ) refers to the total number of carbon atoms in the portion of the heterocyclyl group exclusive of the number of heteroatoms.
  • heterocycle and heteroaryl groups include, but are not limited to, azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, pyridone, indolizine, isoindole, indole, dihydroindole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, naphthalene, oxazole, oxopyrrolidine, piperidine, piperazine
  • cyclopentathiazole thiazolidine, thiophene, benzo[b]thiophene, morpholine, thiomorpholine (also referred to as thiamorpholine), piperidine, pyrrolidine, and tetrahydrofuranyl.
  • fused heterocyclic refers to a 3 to 10 member cyclic substituent formed by the replacement of two hydrogen atoms at different carbon atoms in a cycloalkyl ring structure, as exemplified by the following cyclopentathiazole structure:
  • fused aryl and fused heteroaryl refers to a 5 to 6 member aryl structure or heteroaryl structure fused with a 5- to 6- member aryl, heteroaryl or cycloalkyl ring at different carbon atoms in the aryl structure or the heteroaryl structure, which may be substituted at one of the carbons in the fused aryl or fused heteroaryl and connected to the core molecule at another of the carbons, as exemplified by the following cyclopentylthiazole, quinoline or naphthalene structures:
  • Compound refers to a compound encompassed by the generic formulae disclosed herein, any subgenus of those generic formulae, and any forms of the compounds within the generic and subgeneric formulae, including the racemates, stereoisomers, and tautomers of the compound or compounds.
  • heteroatom means nitrogen, oxygen, or sulfur and includes any oxidized form of nitrogen, such as N(O) ⁇ N + — O " ⁇ and sulfur such as S(O) and S(O) 2 , and the quaternized form of any basic nitrogen.
  • Oxazole and oxazolyl refers to a 5-membered heterocyclic ring containing one nitrogen and one oxygen as heteroatoms and also contains three carbons and may be substituted at one of the three carbons and may be connected to another molecule at another of the three carbons, as exemplified by any of the following structures, wherein the oxazolidinone groups shown here are bonded to a parent molecule, which is indicated by a wavy line in the bond to the parent molecule:
  • Oxopyrrolidine and “oxopyrrolidinyl” refers to a 5-membered heterocyclic ring containing nitrogen and 4 carbons that is substituted at one of the carbons in the heterocyclic ring by a carbonyl and may be connected to another substituent at another carbon in the heterocyclic ring, as exem by the structure below:
  • Pyridine and pyridinyl refers to a 6-membered heteroaryl ring containing one nitrogen and 5 carbons that may also be substituted at one or more of the carbons in the heteroaryl ring, and may be connected to another substituent at another carbon in the heteroaryl ring, as exemplified by the structures below:
  • Thiazole and thiazolyl refers to a 5-membered heteroaryl containing one sulfur and one nitrogen in the heteroaryl ring and 3 carbons in the heteroaryl ring that may also be substituted at one or more of the carbons in the heteroaryl ring, and may be connected to another substituent at another carbon in the heteroaryl ring, as exemplified by the structures belo
  • Pyrimidine and pyrimidinyl refers to a 6-membered heteroaryl ring containing two nitrogens in the heteroaryl ring and 4 carbons in the heteroaryl ring that may be substituted at one or more of the carbons in the heteroaryl ring, and may be connected to another substituent at another carbon in the heteroaryl ring, as exemplified by the structures below:
  • Racemates refers to a mixture of enantiomers.
  • the compounds of Formulas I, or pharmaceutically acceptable salts thereof are enantiomerically enriched with one enantiomer wherein all of the chiral carbons referred to are in one configuration.
  • reference to an enantiomerically enriched compound or salt is meant to indicate that the specified enantiomer will comprise more than 50% by weight of the total weight of all enantiomers of the compound or salt.
  • Solvate or “solvates” of a compound refer to those compounds, as defined above, which are bound to a stoichiometric or non-stoichiometric amount of a solvent.
  • Solvates of a compound includes solvates of all forms of the compound.
  • solvents are volatile, non-toxic, and/or acceptable for administration to humans in trace amounts. Suitable solvates include water.
  • Stereoisomer or “stereoisomers” refer to compounds that differ in the chirality of one or more stereocenters. Stereoisomers include enantiomers and
  • “Pharmaceutically acceptable salt” refers to pharmaceutically acceptable salts derived from a variety of organic and inorganic counter ions well known in the art and include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, and tetraalkylammonium, and when the molecule contains a basic functionality, salts of organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, and oxalate. Suitable salts include those described in P. Heinrich Stahl, Camille G. Wermuth (Eds.), Handbook of Pharmaceutical Salts Properties, Selection, and Use; 2002.
  • Patient refers to mammals and includes humans and non-human mammals.
  • Treating or “treatment” of a disease in a patient refers to 1) preventing the disease from occurring in a patient that is predisposed or does not yet display symptoms of the disease; 2) inhibiting the disease or arresting its development; or 3) ameliorating or causing regression of the disease.
  • dashed line represents an optional double bond at that position.
  • dashed circles appear within ring structures denoted by solid lines or solid circles, then the dashed circles represent one to three optional double bonds arranged according to their proper valence taking into account whether the ring has any optional substitutions around the ring as will be known by one of skill in the art.
  • the dashed line in the structure below could either indicate a double bond at that position or a single bond at that position:
  • ring A below could be a cyclohexyl ring without any double bonds or it could also be a phenyl ring having three double bonds arranged in any position that still depicts the proper valence for a phenyl ring.
  • any of X 1 -X 5 could be selected from: C, CH, or CH 2 , N, or NH, and the dashed circle means that ring B could be a cyclohexyl or phenyl ring or a N-containing heterocycle with no double bonds or a N- containing heteroaryl ring with one to three double bonds arranged in any position that still depicts the proper valen
  • substituents that are not explicitly defined herein are arrived at by naming the terminal portion of the functionality followed by the adjacent functionality toward the point of attachment.
  • substituent "arylalkyloxycarbonyl” refers to the group (aryl)-(alkyl)-0-C(0)-.
  • -CCR' it should be understood that the two R x groups can be the same, or they can be different if R x is defined as having more than one possible identity.
  • certain substituents are drawn as -R x R y , where the "-" indicates a bond adjacent to the parent molecule and R y being the terminal portion of the functionality.
  • impermissible substitution patterns e.g., methyl substituted with 5 fluoro groups. Such impermissible substitution patterns are well known to the skilled artisan.
  • substitution with isotopes such as deuterium, i.e. 2 H may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances.
  • C* is a carbon atom stereocenter which has a configuration which is (R) or (S);
  • R 1 , R 2 , R 3 and R 4 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR 20 R 21 COR 19 ; COOR 19 ; CONR 20 R 21 ; pyrrolidinyl; unsubstituted Ci- 6 alkyl or halo- or alkyl substituted Ci_ 6 alkyl; Ci- 6 alkylCOOR 19 ; -BOR 19 OR 19' ; unsubstituted Ci- 6 alkoxy or halo-or alkyl-substituted Ci- 6 alkyoxy; unsubstituted C 3 - 7 cycloalkyl or halo-or alkyl-substituted C 3 - 7 cycloalkyl; unsubstituted C 3 - 7 cycloalkenyl or halo- or alkyl-substituted C 3 - 7 cycloalkenyl; unsubstituted or halo- or cyano
  • R 5 and R 6 are independently selected from halogen; cyano;
  • R 7 and R 8 are independently hydrogen; halogen; cyano; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- or alkyl- substituted Ci- 6 alkoxy; unsubstituted C 3 - 8 cycloalkyl or halo- or alkyl-substituted C 3 .
  • ecycloalkyl unsubstituted or halo- or alkyl-substituted-C x H 2 x-phenyl; unsubstituted or halo- or alkyl-substituted -0-C x H 2x -phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R 7 and R 8 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR 23 , R 17 , R 17' , R 18 and/or R 18' , or R 7 and R 8 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R 17 , R 17' , R 18 and/or R 18' , wherein a heteroatom in the heterocycloalkyi
  • R 5 and R 8 are independently selected from hydrogen, unsubstituted Ci-6alkyl, and halo- or alkyl-substituted Ci_6alkyl;
  • R 6 and R 7 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR 23 , R 15 , R 15' , R 16 and/or R 16' ; or R 6 and R 7 together form a 3- to 8- membered heterocycloalkyi ring comprising one heteroatom or two or more heteroatoms, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R 15 , R 15' , R 16 and/or R 16' , wherein the one heteroatom in the heterocycloalkyi ring is NR 20 and the two or more heteroatoms are selected from N, NR 22 , O, S, SR 22 and SR 22 R 22' ; or
  • R 5 and R 6 are independently selected from hydrogen; halogen; cyano; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- or alkyl-substituted Ci- 6 alkoxy; unsubstituted C 3 - 8 cycloalkyl or halo- or alkyl- substituted C 3 - 8 cycloalkyl; unsubstituted or halo- or alkyl-substituted -C x H 2x -phenyl;
  • R 5 and R 6 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR 23 , R 17 , R 17' , R 18 and/or R 18' , or R 5 and R 6 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R 13 , R 13 ', R 14 and/or R 14' , wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR 22 , S, SR 22 or SR 22 R 22' ; and
  • R 7 and R 8 are independently selected from halogen; cyano; unsubstituted Ci- 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- or alkyl- substituted Ci- 6 alkoxy; unsubstituted C 3 - 8 cycloalkyl or halo- or alkyl-substituted C 3 .
  • ecycloalkyl unsubstituted or halo- or alkyl-substituted -C x H 2 x-phenyl; unsubstituted or halo- or alkyl-substituted -0-C x H 2 x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; and heteroaryl; or R 7 and R 8 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR 23 , R 17 , R 17' , R 18 and/or R 18' , or R 7 and R 8 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R 17 , R 18 ', R 17 and/or R 18' , wherein a
  • R 9 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- or alkyl- substituted Ci- 6 alkoxy; unsubstituted C 3 . 8 cycloalkyl or halo- or alkyl-substituted C 3 .
  • ecycloalkyl unsubstituted or halo- or alkyl-substituted -C x H 2 x-phenyl; and unsubstituted or halo- or alkyl-substituted-0-C x H 2x -phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
  • R 10 is selected from COOR 19 and CONR 20 R 21 ;
  • R 11 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6 alkyl or halo- or alkyl-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- or alkyl- substituted Ci- 6 alkyoxy; unsubstituted or halo- or alkyl-substituted C 3 .
  • R 12 is selected from hydrogen; unsubstituted Ci_ 6 alkyl or Ci- 6 alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, heterocycloalkyi, heteroaryl, C 3 -8cycloalkyl and C 2 -6alkenyl; Ci-6alkylCi-6alkoxy; Ci-6alkylCi- 6 alkoxyCi- 6 alkoxy; Ci.
  • Ci-6alkynylCi- 6 hydroxy Ci-6alkylCi- 6 alkoxyamino; Ci-6alkylCi-6alkoxyaminoCi- 6 alkyl; Ci_ 6 alkylCi- 6alkoxyaminodiCi- 6 alkyl; Ci- 6 alkylcarboxy; Ci-6alkylaminocarbonylCi- 6 alkoxy; Ci_
  • heteroaryl is an O-containing or N-containing monocyclic heteroaryl
  • R 13 , R 13' , R 14 and R 14' are independently selected from hydrogen; halogen; amino; aminoalkyl; cyano; Ci_ 6 alkyl; Ci- 6 alkoxy; carbonyl; carboxamide; and amide; or R 13 and R 13 ' or R 14 and R 14' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_ 6 alkyl, C 3 -8cycloalkyl, C 2 -6alkenyl or Ci- 6 alkoxy, wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR 22 , S, SR 22 or SR 22 R 22' ;
  • R 15 , R 15' , R 16 and R 16' are independently hydrogen, halogen, amino, cyano, Ci- 6 alkyl, or Ci. 6 alkoxy, wherein any or R 15 , R 15' , R 16 or R 16' , together with R 5 or R 8 , form a 3- to 8-membered cycloalkyl ring or a 3-8 membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_ 6 alkyl, C 3 -8cycloalkyl, C 2 .
  • R 17 , R 17 ', R 18 and R 18' are independently selected from hydrogen; halogen; amino; cyano; Ci_ 6 alkyl; and Ci- 6 alkoxy; or R 17 and R 18 or R 17' and R 18' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_ 6 alkyl, C 3 . 8 cycloalkyl, C 2 .
  • R 19 , R 19' and R 19" are independently selected from hydrogen; Ci_ 6 alkyl; C 3 . ecycloalkyl; C 2 -6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci- 6 alkyltetrazole; Ci- 6 alkylthiazole; Ci- 6 alkyloxazole; Ci- 6 alkyldioxazole; and Ci_
  • R 20 and R 21 are independently selected from hydrogen; Ci_ 6 alkyl; C 3 . ecycloalkyl; C 2 - 6 alkenyl; phenyl; Ci- 6 alkylimidizole; Ci- 6 alkoxy; Ci- 6 alkyltriazole; Ci_
  • R 20 and R 21 together with the nitrogen to which they are attached form unsubstituted pyrrolidinyl, unsubstituted piperidinyl, or unsubstituted morpholinyl; or form carboxyl-substituted pyrrolidinyl, carboxyl-substituted piperidinyl or carboxyl-substituted morpholinyl; and
  • R 22 and R 22' are independently selected from hydrogen; oxygen; Ci_ 6 alkyl; Ci- 6 haloalkyl; C 3 - 8 cycloalkyl; Ci- 6 alkylC 3 - 8 cycloalkyl; C 2 - 6 alkenyl; phenyl; Ci- 6 alkylimidizole; Ci- 6 alkoxy; Ci- 6 alkyltriazole; Ci- 6 alkyltetrazole; Ci- 6 alkylthiazole; Ci- 6 alkyloxazole; Ci_ ealkyldioxazole; Ci- 6 alkyloxazolidone; -COR 19 ; -COOR 19' ; -CSOR 19 " ; and -CONR 20 R 21 ; and
  • R 23 is selected from hydrogen; Ci_ 6 alkyl; C 3 - 8 cycloalkyl; C 2 - 6 alkenyl; phenyl; Ci- 6 alkylimidizole; Ci- 6 alkoxy; Ci- 6 alkyltriazole; Ci- 6 alkyltetrazole; Ci- 6 alkylthiazole; Ci_ 6alkyloxazole; Ci-6alkyldioxazole; Ci-6alkyloxazolidone; aryl; heteroaryl; and benzyl, or a pharmaceutically acceptable salt thereof.
  • One particular embodiment provides a compound according of Formula I, Formula IA or Formula IB as described herein, wherein:
  • R 1 is selected from hydrogen, hydroxy, halogen, cyano, amino, pyrrolidinyl, unsubstituted Ci_ 6 alkyl or halo-substituted Ci_ 6 alkyl, unsubstituted Ci- 6 alkoxy or halo- substituted Ci- 6 alkyoxy; C 3 . 7 cycloalkyl or halo-substituted C 3 .
  • R 2 and R 3 are independently OR 12 ;
  • R 4 is selected from hydrogen, hydroxy, halogen, cyano, amino, pyrrolidinyl, unsubstituted Ci_ 6 alkyl or halo-substituted Ci_ 6 alkyl, unsubstituted Ci- 6 alkoxy or halo- substituted Ci- 6 alkyoxy; C 3 . 7 cycloalkyl or halo-substituted C 3 .
  • R 6 and R 7 together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring comprising one heteroatom or two or more heteroatoms, optionally substituted with R 15 , R 15' , R 16 and/or R 16' , wherein the one heteroatom in the heteroalkyl ring is NR 20 and the two or more heteroatoms are selected from N, NR 22 , O, S, SR 22 and SR 22 R 22' ; R 11 is hydrogen; R 12 is as described herein; and R 15 and R 16 or R 15' and R 16' together form a 3- to 8- membered cycloalkyi ring or heterocycloalkyi ring optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_ 6 alkyl, C 3 - 8 cycloalkyl, C 2 - 6 alkenyl or Ci- 6 alkoxy, wherein the heteroatom in the heteroalkyl ring is
  • Another embodiment provides a compound of Formula I, Formula IA or Formula IB as described herein, wherein R 10 is COOR 19 or CONR 20 R 21 ; or a
  • Another particular embodiment provides a compound of Formula I, Formula IA or Formula IB as described herein, wherein R 10 is COOR 19 ; R 12 is unsubstituted Ci_ 6 alkyl; and R 19 is hydrogen, or a pharmaceutically acceptable salt thereof.
  • Another particular embodiment provides a compound of Formula I, Formula IA or Formula IB as described herein wherein R 4 is hydrogen; R 10 is COOR 19 ; R 12 is -CH 3 ; and R 19 is hydrogen, or a pharmaceutically acceptable salt thereof.
  • Another particular embodiment provides a compound of Formula I, Formula IA or Formula IB as described herein, wherein R 2 and R 3 are independently OR 12 ; R 5 and R 6 together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with R 13 and R 14 , wherein the heteroatom in the heteroalkyl ring is O, N, NR 22 , S, SR 22 or SR 22 R 22 '; R 10 is COOR 19 ; R 11 is hydrogen; R 12 is unsubstituted Ci- 6alkyl; R 13 and R 14 or R 13' and R 14' together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_ 6 alkyl, C 3 - 8 cycloalkyl, C 2 - 6 alkeny
  • Still another particular embodiment provides a compound of Formula I, Formula IA or Formula IB as described herein, wherein R 2 and R 3 are independently OR 12 ; R 7 and R 8 together form a 3- to 8-membered ring, optionally substituted with R 17 and R 18 ; R 10 is COOR 19 ; R 11 is hydrogen; R 12 is unsubstituted Ci_ 6 alkyl; R 17 and R 18 or R 17' and R 18' together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C 3 .
  • ecycloalkyl C 2 - 6 alkenyl or Ci. 6 alkoxy, wherein the heteroatom in the heteroalkyl ring is O, N, NR 22 , S, SR 22 or SR 22 R 22' ; and R 19 is hydrogen, or a pharmaceutically acceptable salt thereof.
  • Another particular embodiment provides a compound of Formula I, Formula IA or Formula IB as described herein, wherein R 2 and R 3 are independently OR 12 ; and R 5 and R 6 together form a 3- to 8- membered cycloalkyl ring or 3- to 8- membered
  • heterocycloalkyl ring optionally substituted with R 13 , R 14 , R 13' and/or R 14' , wherein the heteroatom in the heteroalkyl ring is O, N, NR 22 , S, SR 22 or SR 22 R 22' , or a pharmaceutically acceptable salt thereof.
  • Another particular embodiment provides a compound of Formula I, Formula IA or Formula IB as described herein wherein R 2 and R 3 are independently OR 12 ; and R 7 and R 8 together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered
  • heterocycloalkyl ring optionally substituted with R 17 , R 18 , R 17' and/or R 18' , wherein the heteroatom in the heteroalkyl ring is O, N, NR 22 , S, SR 22 or SR 22 R 22' , or a pharmaceutically acceptable salt thereof.
  • R is H or alkyl
  • compounds of Formula I, Formula IA or Formula IB are selected from:
  • R is hydrogen or alkyi and R' is H, alkyl, ester, carbamate, urea, or sulfonyl urea.
  • R 1 , R 2 , R 3 , R 4 , R 9 , R 10 and R 11 are as described, and R' and R are independently selected from hydrogen, halo, alkyl, aryl, carbocycle, heterocycle, heteroaryl, alkylene optionally substituted with
  • R and R' are independently H, alkyl, acyl, ester, carbamoyl, sulfonyl urea, or urea, and wherein Z is substituted C, O, N or S.
  • R 4 , R 5 , R 6 , R 9 , R 10 and R 11 are as described, and R 1 is halogen, R 2 and R 3 are independently selected from:
  • Compounds described herein can exist in particular geometric or stereoisomeric forms.
  • the invention contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)-isomers, the racemic mixtures thereof, and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, as falling within the scope of the invention.
  • Additional asymmetric carbon atoms can be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in this invention.
  • Optically active (R)- and (S)-isomers and d and I isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. If, for instance, a particular enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis, or by derivatization with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers.
  • diastereomeric salts can be formed with an appropriate optically active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographic means known in the art, and subsequent recovery of the pure
  • enantiomers In addition, separation of enantiomers and diastereomers is frequently accomplished using chromatography employing chiral, stationary phases, optionally in combination with chemical derivatization (e.g., formation of carbamates from amines).
  • chemical derivatization e.g., formation of carbamates from amines.
  • a pharmaceutical composition comprising a pharmaceutically acceptable diluent and a therapeutically effective amount of a compound of Formula I, Formula IA or Formula IB, or a compound selected from Table 1 A and Table 1 B, or a pharmaceutically acceptable salt thereof.
  • the pharmaceutical formulation containing a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1 A and Table 1 B or a pharmaceutically acceptable salt thereof is a formulation adapted for parenteral administration.
  • the formulation is a long-acting parenteral formulation.
  • the formulation is a nano-particle formulation.
  • the pharmaceutical formulation containing a compound of Formula I, Formula IA or Formula IB, or a compound selected from Table 1 A and Table 1 B, or a pharmaceutically acceptable salt thereof is a formulation adapted for oral, rectal, topical or intravenous formulation, wherein the pharmaceutical formulation optionally comprises any one or more of a pharmaceutically acceptable carrier, adjuvant or vehicle.
  • the compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1A and Table 1 B or a pharmaceutically acceptable salt thereof is formulated for oral administration, and can be administered as a conventional preparation, for example, as any dosage form of a solid agent such as tablets, powders, granules, capsules and the like; an aqueous agent; an oily suspension; or a liquid agent such as syrup and elixir.
  • the compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1 A and Table 1 B or a pharmaceutically acceptable salt thereof is formulated for parenteral administration, and can be administered as an aqueous or oily suspension injectable, or a nasal drop.
  • a preparation of a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1A and Table 1 B or a pharmaceutically acceptable salt thereof may be prepared by combining (e.g. mixing) a therapeutically effective amount of a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1 A and Table 1 B or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable carrier or diluent.
  • compositions adapted for oral administration may be presented as discrete units such as capsules or tablets; powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; edible foams or whips; or oil-in-water liquid emulsions or water-in-oil liquid emulsions.
  • the compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1A and Table 1 B can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like.
  • Powders are prepared by comminuting the compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1 A and Table 1 B to a suitable fine size and mixing with a similarly comminuted pharmaceutical carrier such as an edible carbohydrate, as, for example, starch or mannitol. Flavoring, preservative, dispersing and coloring agent can also be present.
  • Capsules are made by preparing a powder mixture, as described above, and filling formed gelatin sheaths.
  • Glidants and lubricants such as colloidal silica, talc, magnesium stearate, calcium stearate or solid polyethylene glycol can be added to the powder mixture before the filling operation.
  • a disintegrating or solubilizing agent such as agar-agar, calcium carbonate or sodium carbonate can also be added to improve the availability of the medicament when the capsule is ingested.
  • suitable binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture.
  • Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like.
  • Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like.
  • Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. Tablets are formulated, for example, by preparing a powder mixture, granulating or slugging, adding a lubricant and disintegrant and pressing into tablets.
  • a powder mixture is prepared by mixing the compound, suitably comminuted, with a diluent or base as described above, and optionally, with a binder such as carboxymethylcellulose, an aliginate, gelatin, or polyvinyl pyrrolidone, a solution retardant such as paraffin, a resorption accelerator such as a quaternary salt and/or an absorption agent such as bentonite, kaolin or dicalcium phosphate.
  • the powder mixture can be granulated by wetting with a binder such as syrup, starch paste, acadia mucilage or solutions of cellulosic or polymeric materials and forcing through a screen.
  • the powder mixture can be run through the tablet machine and the result is imperfectly formed slugs broken into granules.
  • the granules can be lubricated to prevent sticking to the tablet forming dies by means of the addition of stearic acid, a stearate salt, talc or mineral oil.
  • the lubricated mixture is then compressed into tablets.
  • the compounds of the present invention can also be combined with a free flowing inert carrier and compressed into tablets directly without going through the granulating or slugging steps.
  • a clear or opaque protective coating consisting of a sealing coat of shellac, a coating of sugar or polymeric material and a polish coating of wax can be provided. Dyestuffs can be added to these coatings to distinguish different unit dosages.
  • Oral fluids such as solutions, syrups and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of the compound.
  • Syrups can be prepared by dissolving the compound in a suitably flavored aqueous solution, while elixirs are prepared through the use of a non-toxic alcoholic vehicle.
  • Suspensions can be formulated by dispersing the compound in a non-toxic vehicle.
  • Solubilizers and emulsifiers such as ethoxylated isostearyl alcohols and polyoxy ethylene sorbitol ethers, preservatives, flavor additive such as peppermint oil or natural sweeteners or saccharin or other artificial sweeteners, and the like can also be added.
  • dosage unit formulations for oral administration can be microencapsulated.
  • the formulations of compounds of Formula I, Formula IA or Formula IB can also be prepared to prolong or sustain the release of the compound, as for example by coating or embedding particulate material in polymers, wax or the like.
  • the compounds of Formula I, Formula IA or Formula IB or compounds selected from Table 1A and Table 1 B, or salts, solvates or hydrates thereof, can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles.
  • liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.
  • the compounds of Formula I, Formula IA or Formula IB or compounds selected from Table 1 A and Table 1 B, or salts, solvates or hydrates thereof, may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled.
  • the compounds may also be coupled with soluble polymers as targetable drug carriers.
  • Such polymers can include polyvinylpyrrolidone, pyrancopolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxyethylaspartamide-phenol, or polyethyleneoxidepolylysine substituted with palmitoyl residues.
  • the compounds may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked or amphipathic block copolymers of hydrogels.
  • a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked or amphipathic block copolymers of hydrogels.
  • compositions adapted for transdermal administration may be presented as discrete patches intended to remain in intimate contact with the epidermis of the recipient for a prolonged period of time.
  • the compounds of Formula I, Formula IA or Formula IB or compounds selected from Table 1 A and Table 1 B may be delivered from a patch by iontophoresis as generally described in Pharmaceutical Research, 3(6), 318 (1986).
  • compositions adapted for topical administration may be formulated as ointments, creams, suspensions, lotions, powders, solutions, pastes, gels, sprays, aerosols or oils.
  • the active ingredient When formulated in an ointment, the active ingredient may be employed with either a paraffinic or a water-miscible ointment base.
  • the active ingredient may be formulated in a cream with an oil-in-water cream base or a water-in-oil base.
  • compositions adapted for rectal administration may be presented as suppositories or as enemas.
  • compositions adapted for nasal administration wherein the carrier is a solid include a coarse powder having a particle size for example in the range 20 to 500 microns which is administered in the manner in which snuff is taken, i.e. by rapid inhalation through the nasal passage from a container of the powder held close up to the nose.
  • Suitable formulations wherein the carrier is a liquid, for administration as a nasal spray or as nasal drops, include aqueous or oil solutions of the active ingredient.
  • Fine particle dusts or mists which may be generated by means of various types of metered, dose pressurized aerosols, nebulizers or insufflators.
  • compositions adapted for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents.
  • the formulations may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use.
  • Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets.
  • formulations described herein may include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavouring agents.
  • a therapeutically effective amount of a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1A and Table 1 B will depend upon a number of factors including, for example, the age and weight of the human or other animal, the precise condition requiring treatment and its severity, the nature of the formulation, and the route of administration, and will ultimately be at the discretion of the attendant physician or veterinarian.
  • An effective amount of a salt or hydrate thereof may be determined as a proportion of the effective amount of the compound of Formula I, Formula IA or Formula IB or the compound selected from Table 1 A and Table 1 B, or salts, solvates or hydrates thereof, per se.
  • Embodiments of the present invention provide administration of a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1 A and Table 1 B to a healthy or virus-infected patient, either as a single agent or in combination with (a) another agent that is effective in treating or preventing hepatitis B virus of hepatitis D virus, (b) another agent that improves immune response and robustness, or (c) another agent that reduces inflammation and/or pain.
  • a method of treating a hepatitis B virus and/or hepatitis D virus by administering a therapeutically effective amount of a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1 A and Table 1 B, or a pharmaceutically acceptable salt, solvate or hydrate thereof, to a mammal.
  • the mammal is a human.
  • a method of reducing HBe and/or HBs antigens in a mammal by administering to said mammal a therapeutically effective amount of a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1A and Table 1 B or a pharmaceutically acceptable salt, solvate or hydrate thereof.
  • the mammal is a human.
  • the compounds of the present invention may be used in combination with one or more antiviral therapeutic agents or anti-inflammatory agents useful in the prevention or treatment of viral diseases or associated pathophysiology.
  • the compounds of the present invention and their salts, solvates, or other pharmaceutically acceptable derivatives thereof may be employed alone or in combination with other antiviral or anti-inflammatory therapeutic agents.
  • the compounds of the present invention and any other pharmaceutically active agent(s) may be administered together or separately and, when administered separately, administration may occur simultaneously or sequentially, in any order.
  • the amounts of the compounds of the present invention and the other pharmaceutically active agent(s) and the relative timings of administration will be selected in order to achieve the desired combined therapeutic effect.
  • the administration in combination of a compound of the present invention and salts, solvates, or other pharmaceutically acceptable derivatives thereof with other treatment agents may be in combination by administration concomitantly in: (1) a unitary pharmaceutical composition including both compounds; or (2) separate pharmaceutical compositions each including one of the compounds.
  • the combination may be administered separately in a sequential manner wherein one treatment agent is administered first and the other second or vice versa. Such sequential administration may be close in time or remote in time.
  • the amounts of the compound(s) of Formula I, Formula IA or Formula IB or compounds selected from Table 1 A and Table 1 B or salts thereof, and the other pharmaceutically active agent(s), and the relative timings of administration, will be selected in order to achieve the desired combined therapeutic effect.
  • embodiments provide a method as described comprising administering an additional agent selected from an antiviral agent, an antibiotic, an analgesic, a non-steroidal anti-inflammatory (NSAID) agent, an antifungal agent, an antiparasitic agent, an anti-nausea agent, an anti-diarrheal agent, or an immunosuppressant agent.
  • the antiviral agent is an anti-hepatitis B agent or an anti- hepatitis C agent.
  • the additional agent is administered as part of a single dosage form of said pharmaceutical formulation, or as a separate dosage form.
  • the present invention is directed to compounds, compositions and pharmaceutical compositions that have utility as novel treatments and/or preventative therapies for virus infections. While not wanting to be bound by any particular theory, it is thought that the present compounds are able to inhibit the levels of HBe and HBs antigens in a subject infected with hepatitis B virus or suffering from a chronic hepatitis B viral infection. By reducing the levels of HBe and HBs antigens in a subject infected with hepatitis B virus, compounds described herein are effective at treating hepatitis B infections, and secondary disorders such as liver cirrhosis, liver failure and liver cancer which are often associated with hepatitis B virus infections.
  • a method of treating or preventing a hepatitis B virus infection in a subject suffering from the HBV infection comprising administering to the subject an inhibitor of HBe and/or HBs antigens, wherein the inhibitor is a compound of Formula I, Formula IA or Formula IB.
  • a method of treating a hepatitis B virus infection and/or a hepatitis D virus infection in a subject suffering from the virus infection comprising administering to the subject a compound from Table 1 A or a compound from Table 1 B .
  • a method for treating a viral infection in a subject mediated at least in part by a virus in the hepatitis B family and or the hepatitis D family comprising administering to the subject a composition comprising a compound of any of Formula I, Formula IA or Formula IB, or a compound selected from Table 1 A and Table 1 B, or a pharmaceutically acceptable salt thereof.
  • another embodiment of the present invention provides a method of inhibiting progression of a viral infection in a subject at risk for infection with a hepatitis B virus and/or a hepatitis D virus, comprising administering to the subject a therapeutically effective amount of the compound of Formula I, Formula IA or Formula IB, or a compound selected from Table 1A and Table 1 B, or a pharmaceutically acceptable salt thereof.
  • another embodiment of the present invention provides a method of preventing a viral infection in a subject at risk for infection from a hepatitis B virus and/or a hepatitis D virus comprising administering to the subject a therapeutically effective amount of the compound of Formula I or Formula IA, or a compound selected from Table 1 A and Table 1 B, or a pharmaceutically acceptable salt thereof.
  • another embodiment of the present invention provides a method of treating a virus infection in a subject suffering from said virus infection, comprising administering to the subject a therapeutically effective amount of the compound of Formula I, Formula IA or Formula IB, or a compound selected from Table 1 A and Table 1 B, or a pharmaceutically acceptable salt thereof.
  • Compounds, methods and pharmaceutical compositions for treating viral infections, by administering a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1A and Table 1 B in a therapeutically effective amount are disclosed.
  • Methods for preparing compounds of Formula I, Formula IA or Formula IB or compounds selected from Table 1A and Table 1 B and methods of using the compounds and pharmaceutical compositions thereof are also disclosed.
  • the treatment and prophylaxis of viral infections such as those caused by hepatitis B and/or hepatitis D are disclosed.
  • the compounds described herein are useful for treating infections in a subject wherein the infection is caused by a multi-drug resistant strain of the hepatitis B virus and/or a hepatitis D virus.
  • the compound of the present invention is chosen from the compounds set forth in Table 1A and Table 1 B.
  • C* is a carbon atom stereocenter which has a configuration which is (R) or
  • R 1 , R 2 , R 3 and R 4 are independently selected from hydrogen, hydroxy, halogen, cyano, amino, pyrrolidinyl, unsubstituted Ci_ 6 alkyl or halo-substituted Ci_ 6 alkyl, unsubstituted Ci- 6 alkoxy or halo-substituted Ci- 6 alkyoxy; C 3 - 7 cycloalkyl or halo-substituted C 3 - 7 cycloalkyl; N-containing monocyclic heterocycloalkyi, pyrrolidinyl, -C x H 2x -phenyl, -O- C x H 2x -phenyl or -(Ci-6alkyl)N- C x H 2x -phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; or -OR 12 ; and wherein
  • R 5 and R 6 are independently halogen, cyano, unsubstituted Ci_ 6 alkyl or halo-substituted Ci_ 6 alkyl, unsubstituted Ci- 6 alkoxy or halo-substituted Ci- 6 alkoxy, unsubstituted C 3 - 8 cycloalkyl or halo-substituted C 3 - 8 cycloalkyl, -C x H 2 x-phenyl or -0-C x H 2x - phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; heteroaryl; or R 5 and R 6 together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with R 13 , R 13' , R 14 and/or R 14' , wherein the heteroatom in the heterocycloalkyi ring is
  • R 7 and R 8 are independently hydrogen, halogen, cyano, unsubstituted Ci_ 6 alkyl or halo-substituted Ci_ 6 alkyl, unsubstituted Ci- 6 alkoxy or halo-substituted Ci- 6 alkoxy, unsubstituted C 3 . 8 cycloalkyl or halo-substituted C 3 .
  • R 5 is selected from hydrogen, unsubstituted Ci_ 6 alkyl, and halo- or alkyl-substituted Ci_ 6 alkyl;
  • R 6 and R 7 together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring comprising one heteroatom or two or more heteroatoms, optionally substituted with R 15 , R 15' , R 16 and/or R 16' , wherein the one heteroatom in the heteroalkyl ring is NR 20 and the two or more heteroatoms are selected from N, NR 22 , O, S, SR 22 and SR 22 R 22' ; and
  • R 8 is selected from hydrogen, unsubstituted Ci_ 6 alkyl, and halo- or alkyl- substituted Ci- 6 alkyl; or
  • R 5 and R 6 are independently hydrogen, halogen, cyano,
  • Ci_ 6 alkyl or halo-substituted Ci_ 6 alkyl unsubstituted Ci- 6 alkoxy or halo- substituted Ci- 6 alkoxy, unsubstituted C 3 . 8 cycloalkyl or halo-substituted C 3 . 8 cycloalkyl, - C x H 2x -phenyl or -0-C x H 2x -phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; heteroaryl; or R 5 and R 6 together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered
  • heterocycloalkyi ring optionally substituted with R 13 , R 13 ', R 14 and/or R 14' , wherein the heteroatom in the heterocycloalkyi ring is selected from O, N, NR 22 , S, SR 22 or SR 22 R 22' ; and
  • R 7 and R 8 are independently halogen, cyano, unsubstituted Ci_ 6 alkyl or halo- substituted Ci- 6 alkyl, unsubstituted Ci- 6 alkoxy or halo-substituted Ci- 6 alkoxy, unsubstituted C 3 - 8 cycloalkyl or halo-substituted C 3 - 8 cycloalkyl, -C x H 2 x-phenyl or -0-C x H 2x -phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; heteroaryl; or R 7 and R 8 together form a 3- to 8- membered cycloalkyi ring or a 3- to 8- membered heterocycloalkyi ring, optionally substituted with R 17 , R 18 ', R 17 and/or R 18' , wherein the heteroatom in the heterocycloalkyi ring is selected from O, N
  • R 9 is hydrogen, hydroxy, halogen, cyano, unsubstituted Ci_ 6 alkyl or halo- substituted Ci- 6 alkyl, unsubstituted Ci- 6 alkoxy or halo-substituted Ci- 6 alkoxy, unsubstituted C 3 - 8 cycloalkyl or halo-substituted C 3 - 8 cycloalkyl, -C x H 2 x-phenyl or -0-C x H 2x -phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6;
  • R 10 is hydrogen; hydroxy; halogen; cyano; COOR 19 , CONR 20 R 21 , unsubstituted Ci_ 6 alkyl or halo-substituted Ci_ 6 alkyl; unsubstituted Ci- 6 alkoxy or halo- substituted Ci- 6 alkyoxy; C 3 . 7 cycloalkyl; -C x H 2 x-phenyl or -0-C x H 2x -phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6;
  • R 11 is hydrogen, hydroxy, halogen, cyano, unsubstituted Ci_ 6 alkyl or halo- substituted Ci- 6 alkyl, unsubstituted Ci- 6 alkoxy or halo-substituted Ci- 6 alkyoxy; C 3 . 7 cycloalkyl; -C x H 2x -phenyl or -0-C x H 2x -phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6;
  • R 12 is hydrogen; unsubstituted Ci_ 6 alkyl or Ci_ 6 alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, C 3 . 8 cycloalkyl and C 2 . 6alkenyl; Ci- 6 alkylCi- 6 alkoxy; Ci- 6 alkylCi- 6 alkoxyCi- 6 alkoxy; Ci- 8 alkylamino; Ci_
  • R 13 , R 13' , R 14 and R 14' are independently hydrogen, halogen, amino, aminoalkyl, cyano, Ci_ 6 alkyl, Ci- 6 alkoxy, carbonyl, carboxamide, amide; or R 13 and R 13 ' or R 14 and R 14' together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci- 6 alkyl, C 3 - 8 cycloalkyl, C 2 - 6 alkenyl or Ci- 6 alkoxy, wherein the heteroatom in the
  • heterocycloalkyi ring is selected from O, N, NR 22 , S, SR 22 or SR 22 R 22' ;
  • R 15 , R 15' , R 16 and R 16' are independently hydrogen, halogen, amino, cyano, Ci- 6 alkyl, or Ci- 6 alkoxy, wherein any of R 15 , R 15' , R 16 or R 16' , together with R 5 or R 8 , form a 3- to 8-membered cycloalkyi ring or a 3-8 membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_ 6 alkyl, C 3 - 8 cycloalkyl, C 2 .
  • R 17 , R 17 ', R 18 and R 18' are independently hydrogen, halogen, amino, cyano, Ci- 6 alkyl, or Ci- 6 alkoxy; or R 17 and R 18 or R 17' and R 18' together form a 3- to 8-membered cycloalkyi or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_ 6 alkyl, C 3 - 8 cycloalkyl, C 2 - 6 alkenyl or Ci- 6 alkoxy, wherein the heteroatom in the heterocycloalkyi ring is selected from O, N, NR 22 , S, SR 22 or SR 22 R 22' ;
  • R 19 , R 19' and R 19 " are independently hydrogen, Ci_ 6 alkyl, C 3 - 8 cycloalkyl, C 2 . 6 alkenyl, phenyl, Ci- 6 alkylimidizole, Ci- 6 alkoxy, Ci- 6 alkyltriazole, Ci- 6 alkyltetrazole, Ci_
  • R 20 and R 21 are independently hydrogen, Ci_ 6 alkyl, C 3 - 8 cycloalkyl, C 2 . 6 alkenyl, phenyl, Ci- 6 alkylimidizole, Ci- 6 alkoxy, Ci- 6 alkyltriazole,Ci- 6 alkyltetrazole, Ci_
  • R 22 and R 22' are independently selected from hydrogen, oxygen, Ci_ 6 alkyl, C 3 -ecycloalkyl, C 2 -6alkenyl, phenyl, Ci-6alkylimidizole, Ci_6alkoxy, Ci-6alkyltriazole,Ci- 6 alkyltetrazole, Ci- 6 alkylthiazole, Ci- 6 alkyloxazole, Ci- 6 alkyldioxazole; Ci- 6 alkyloxazolidone, - COR 19 , -COOR 19' , -CSOR 19 " , -CONR 20 R 21 , or a pharmaceutically acceptable salt thereof.
  • One particular embodiment provides a compound according of Formula I, Formula IA or Formula IB as described herein, wherein:
  • R 1 is selected from hydrogen, hydroxy, halogen, cyano, amino, pyrrolidinyl, unsubstituted Ci_ 6 alkyl or halo-substituted Ci_ 6 alkyl, unsubstituted Ci- 6 alkoxy or halo- substituted Ci- 6 alkyoxy; C 3 . 7 cycloalkyl or halo-substituted C 3 .
  • R 2 and R 3 are independently OR 12 ;
  • R 4 is selected from hydrogen, hydroxy, halogen, cyano, amino, pyrrolidinyl, unsubstituted Ci_ 6 alkyl or halo-substituted Ci_ 6 alkyl, unsubstituted Ci- 6 alkoxy or halo- substituted Ci- 6 alkyoxy; C 3 - 7 cycloalkyl or halo-substituted C 3 - 7 cycloalkyl; N-containing monocyclic heterocycloalkyl, pyrrolidinyl, -C x H 2x -phenyl, -0-C x H2x-phenyl, or -(Ci_ 6 alkyl)N- C x H 2 x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; or -OR 12 ;
  • R 6 and R 7 together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyl ring, optionally substituted with R 15 , R 15 ', R 16 and/or R 16' , wherein the heteroatom in the heteroalkyl ring is O, N, NR 22 , S, SR 22 or SR 22 R 22' ; R 9 is hydrogen; R 11 is hydrogen; R 12 is as described herein; and R 15 and R 16 or R 15' and R 16' together form a 3- to 8-membered cycloalkyl ring or heterocycloalkyl ring optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_ 6 alkyl, C 3 - 8 cycloalkyl, C 2 . 6 alkenyl or Ci- 6 alkoxy, wherein the heteroatom in the heteroalkyl ring is O, N, NR 22 , S, SR 22 or SR 22
  • Suitable synthetic routes are depicted below in the following general reaction schemes.
  • the skilled artisan will appreciate that if a substituent described herein is not compatible with the synthetic methods described herein, the substituent may be protected with a suitable protecting group that is stable to the reaction conditions.
  • the protecting group may be removed at a suitable point in the reaction sequence to provide a desired intermediate or target compound.
  • Suitable protecting groups and the methods for protecting and de-protecting different substituents using such suitable protecting groups are well known to those skilled in the art; examples of which may be found in T. Greene and P. Wuts, Protecting Groups in Chemical Synthesis (3rd ed.), John Wiley & Sons, NY (1999).
  • a substituent may be specifically selected to be reactive under the reaction conditions used. Under these circumstances, the reaction conditions convert the selected substituent into another substituent that is either useful as an intermediate compound or is a desired substituent in a target compound.
  • DIPEA or DIEA
  • ⁇ , ⁇ -diisopropylethylamine or Hiinig's base
  • T3P 1 -Propanephosphonic anhydride solution 2,4,6-Tripropyl-1 ,3,5,2,4,6- trioxatriphosphorinane-2,4,6-trioxide
  • Flash chromatography was run over Merck Silica gel 60 (230 - 400 mesh), or using a Teledyne Isco Combiflash Companion with normal phase, disposable Redi-Sep flash columns.
  • Mass spectra were run on an open access LC-MS system using electrospray ionization.
  • the analytical low-resolution mass spectra (MS) were recorded on Waters SQD instrument with UPLC analysis was conducted on a Phenomenex Kinetex 1 .7um, 2.1 x 50mm XB-C18 column at 40CSQ using a gradient elution method.
  • Solvent A 0.2% formic acid (FA) in water
  • Solvent B 0.15% FA in acetonitrile
  • 1 % - 99% Solvent B gradient over 1 .1 minutes and holding steady at 99% solvent B for another 0.4 minutes, at 1 ml/min flow rate.
  • MS analytical low-resolution mass spectra
  • Solvent A 0.1 % formic acid (FA) in water
  • Solvent B 0.1 % FA in acetonitrile
  • compound 1 can be prepared by reacting 4-bromo-1 ,2-dimethoxybenzene 9 with a ketone, as shown, using a palladium/xantphos Buchwald-Hartwig chemistry in sodium t-butoxide and THF. Reductive amination of 1 by reaction of compound 1 with sodium cyanoborohydride in ammonium acetate follows, to yield amine 2. Amine 2 is then reacted with formic acid in reflux conditions to yield amide 3. The dihydroisoquinoline compound 4 is obtained by a cyclization/condensation reaction using phosphoryl chloride in acetonitrile.
  • R 15 , R 15' ' R 16 and R 16' are as described herein.
  • compound 10 can be prepared by reacting 4-bromo-1 ,2- dimethoxybenzene 9 with a ketone, as shown, using a palladium/xantphos Buchwald- Hartwig chemistry in sodium t-butoxide and THF. Reductive amination of 10 by reaction of compound 10 with sodium cyanoborohydride in ammonium acetate follows, to yield amine 11. Amine 11 is then reacted with formic acid in reflux conditions to yield amide 12. The dihydroisoquinoline compound 13 is obtained by a cyclization/condensation reaction using phosphoryl chloride in acetonitrile.
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 amd R 8 are as described herein.
  • Step 1 Benzyl 3-(benzyloxy)-4-bromobenzoate
  • Step 5 romobenzyl)cvclobutane-1 -carboxylic acid
  • Step 6 bromobenzyl)cvclobutan-1 -amine
  • Step 8 '-(Benzyloxy)-7'-bromo-4'H-spiro[cvclobutane-1 ,3'-isoquinoline
  • Step 9 Ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-1 '.2'.7'.1 1 b 1 - tetrahydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 10 Ethyl 9'-(benzyloxy)-10'-bromo ⁇ '-oxo ⁇ 'J 1 - dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 1 1 9'-(Benzyloxy)-10'-bromo-2'-oxo-2',7'-dihvdrospiro[cvclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • Step 1 Ethyl 10'-bromo-9'-hvdroxy-2'-oxo-2',7'-dihvdrospiro[cvclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 2 Ethyl 10'-bromo-9'-(cvclopropylmethoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 3 Ethyl 10'-acetyl-9'-(cvclopropylmethoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 4 Ethyl 9'-(cvclopropylmethoxy)-10'-(2-methyl-1 .3-dithiolan-2-yl)-2'- oxo-2', 7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 5 Ethyl 9'-(cvclopropylmethoxy)-1 Q'-(1 .1 -difluoroethyl)-2'-oxo-2'.7'- dihydrospirofcyclobuta '-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 6 9'-(Cvclopropylmethoxy)-10'-(1 .1 -difluoroethyl)-2'-oxo-2'.7'- dihydrospirofcyclobuta '-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • Step 1 Ethyl 9'-(cvclopropylmethoxyV2'-oxo-10'-vinyl-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 2 Ethyl 9'-(cvclopropylmethoxy)-10'-ethyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 3 9'-(Cvclopropylmethoxy)-10'-ethyl-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
  • Step 1 Ethyl 9'-(cvclopropylmethoxy)-2'-oxo-10'-vinyl-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 2 Ethyl 9'-(cvclopropylmethoxy)-10'-formyl-2'-oxo-2'.7'- dihydrospirofcyclo
  • Step 3 Ethyl 9'-(cvclopropylmethoxy)-10'-(difluoromethyl)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 4 9'-(Cvclopropylmethoxy)-10'-(difluoromethyl)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
  • Step 1 Ethyl 9'-(cvclopropylmethoxy)-10'-(hvdroxymethyl)-2'-oxo-2',7'- dihydrospiro [cyclobuta '-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 2 Ethyl 9'-(cvclopropylmethoxyV10'-(fluoromethyr)-2'-oxo-2'.7'- dihydrospiro [cyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 3 9'-(Cvclopropylmethoxy)-10'-(fluoromethyl)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • Step 1 Ethyl 9'-(cvclopropylmethoxyV2'-oxo-10'-(trifluoromethvn-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 2 9'-(Cvclopropylmethoxy)-2'-oxo-10'-(trifluoromethyl)-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • n-Butyllithium (2.5 M in hexanes) (0.581 mL, 1 .45 mmol) was added dropwise to a -78 °C solution of diisopropylamine (0.207 mL, 1 .45 mmol) in tetrahydrofuran (THF) (5 mL). The mixture was allowed to warm to room temperature and stirred for 15 minutes. The mixture was cooled to -78 °C before methyl tetrahydro-2H-pyran-4- carboxylate (0.185 mL, 1 .38 mmol) was added dropwise. The mixture was warmed to 0 °C and stirred for 15 minutes. A white precipitate formed.
  • THF tetrahydrofuran
  • Step 2 4-(4-Methoxy-3-(3-methoxypropoxy)benzyl)tetrahvdro-2H-pyran-4- carboxylic acid
  • Step 3 4-(4-Methoxy-3-(3-methoxypropoxy)benzyl)tetrahvdro-2H-pyran-4- amine
  • Step 4 N-(4-(4-Methoxy-3-(3-methoxypropoxy)benzyl)tetrahvdro-2H-pyran- 4-yl)formamide
  • Step 5 7-Methoxy-6-(3-methoxypropoxy)-2',3',5',6'-tetrahydro-4H-
  • Step 6 Ethyl 10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2.2'.3.5.6.7'- hexahvdrospiro[pyran-4,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 7 10'-Methoxy-9'-(3-methoxypropoxy)-2'-oxo-2.2'.3.5.6.7'- hexahvdrospiro[pyran-4,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • Step 1 Methyl 1 -(4-chloro-3-methoxybenzyl)cvclohexanecarboxylate
  • n-Butyllithium (2.5 M in hexanes) (6.06 mL, 15.2 mmol) was added dropwise to a -78 °C solution of diisopropylamine (2.16 mL, 15.2 mmol) in tetrahydrofuran (THF) (40 mL). The mixture was allowed to warm to room temperature and stirred for 15 minutes. The mixture was cooled to -78 °C before methyl cyclohexanecarboxylate (2.06 mL, 14.4 mmol) was added dropwise. The mixture was warmed to 0 °C and stirred for 15 minutes.
  • THF tetrahydrofuran
  • Step 2 -(4-Chloro-3-methoxybenzyl)cvclohexanecarboxylic acid
  • Step 3 1 -(4-Chloro-3-methoxybenzyl)cvclohexanamine
  • Diphenyl phosphorazidate (3.84 ml_, 17.82 mmol) was added dropwise to a 0 °C stirring mixture of 1 -(4-chloro-3-methoxybenzyl)cyclohexane-1 -carboxylic acid (2.29 g, 8.10 mmol) and triethylamine (2.483 ml_, 17.82 mmol) in toluene (40 ml_). The mixture was allowed to warm to room temperature and stirred for 15 minutes before being heated at 80 °C for 1 hour.
  • the mixture was allowed to cool to room temperature before a mixture 5N hydrogen chloride (40 ml_, 200 mmol) and 1 ,4-dioxane (40 ml_) was added.
  • the mixture was heated at 80 °C with vigorous stirring for 1 hour.
  • the mixture was allowed to cool to room temperature and was extracted with ethyl ether.
  • the ethyl ether layer was back-extracted 2 times with 1 N hydrochloric acid.
  • the combined acidic aqueous layers were basified with 1 M sodium hydroxide and extracted 3 times with ethyl acetate.
  • Step 4 N-(1 -(4-chloro-3-methoxybenzyl)cvclohexyl)formamide
  • Step 5 7'-Chloro-6'-methoxy-4'H-spiro[cvclohexane-1 ,3'-isoquinolinel
  • Step 6 Ethyl 10'-chloro-9'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclohexane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 7 10'-Chloro-9'-hvdroxy-2'-oxo-2',7'-dihvdrospiro[cvclohexane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • the mixture was poured into ice, diluted with 1 M hydrochloric acid, and extracted 3 times with ethyl acetate. The combined organic layers were concentrated. The residue was dissolved in methanol (5 mL), basified with 1 M sodium hydroxide (10 ml_), and heated at 70 C for 3 hours. The mixture was allowed to cool to room temperature, acidified with 1 N hydrochloric acid, and extracted 3 times with ethyl acetate.
  • Step 8 10'-Chloro-9'-(cvclopropylmethoxy)-2'-oxo-2'.7'- dihydrospirofcyclohexa -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • the mixture was allowed to cool to room temperature, acidified with 1 N hydrochloric acid, and a precipitate collected by filtration.
  • the product was slurried in acetonitrile, sonicated, and the precipitate collected by filtration.
  • the sample was air dried.
  • the crude product was purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient).
  • Step 1 Ethyl 9'-(cvclopropylmethoxy)-10'-methyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 2 9'-(CvclopropylmethoxyV10'-methyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • Step 1 Ethyl 9'-(benzyloxy)-10'-methyl ⁇ '-oxo ⁇ 'J 1 - dihvdrospiro[cvclobutan '-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 2 Ethyl 9'-hvdroxy-10'-methyl-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 3 Ethyl 10'-methyl-2'-oxo-9'-(((trifluoromethyl)sulfonyl)oxy)-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 4 Ethyl 10'-methyl ⁇ '-oxo-g'-foyrrolidin-l -yl)-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 5 10'-Methyl-2'-oxo-9'-(pyrrolidin-1 -yl)-2',7'-dihvdrospiro[cyclobutane-
  • Step 1 Ethyl 10'-methyl-9'-(oxetan-3-ylmethoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 2 10'-Methyl-9'-(oxetan-3-ylmethoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • Step 1 Ethyl 9'-(cvclopentylmethoxy)-10'-methyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 2 9'-(Cvclopentylmethoxy)-10'-methyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • Stepl Ethyl 10'-methoxy-9'-f 1 -methyl-1 H-pyrazol ⁇ -vD ⁇ '-oxo ⁇ 'J'- dihydrospiro
  • Step 2 10'-Methoxy-9'-(1 -methyl-1 H-pyrazol-4-yl)-2'-oxo-2'.7'-dihvdrospiro
  • Stepl Ethyl 9'-(furan-3-yl)-10'-methoxy ⁇ '-oxo ⁇ 'J 1 - dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
  • Step 2 9'-(Furan-3-yl)-10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • Step 1 10'-Chloro-9'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclohexane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • Step 1 (Tetrahydro-2/-/-pyran-4-yl)methyl 4-methylbenzenesulfonate
  • Step 2 Ethyl 9'-(benzyloxy)-2'-oxo-10'-vinyl-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 3 Ethyl 9'-(benzyloxy)-10'-formyl-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 4 Ethyl 9'-(benzyloxy)-10'-toifluoromethyl ⁇ '-oxo ⁇ 'J 1 - dihvdrospiro[cvclobutan -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
  • Step 5 Ethyl 1 O'-toifluoromethviyg'-hvdroxy ⁇ '-oxo ⁇ 'J 1 - dihydrospirofcyclobutan -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
  • Step 6 Ethyl 10'-(difluoromethyl)-2'-oxo-9'-((tetrahvdro-2H-pyran-4- yl)methoxy)-2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
  • Step 7 10'-(Difluoromethyl)-2'-oxo-9'-((tetrahydro-2H-pyran-4-yl)methoxy)- 2',7'-dihvdrospiro[cyclobutane- '-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • Example 17 [00432] 10'-(DifluoromethylV2'-oxo-9'-((tetrahvdrofuran-3-vnmethoxyV2'.7'-)
  • Step 3 Ethyl 10'-formyl-2'-oxo-9'-((tetrahvdrofuran-3-yl)methoxyV2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 4 Ethyl 10'-(difluoromethyl)-2'-oxo-9'-((tetrahydrofuran-3-yl)methoxy)- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 5 10'-(DifluoromethylV2'-oxo-9'-((tetrahvdrofuran-3-yl)methoxyV2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • Step 1 3-Hvdroxy-3-methylbutyl 4-methylbenzenesulfonate
  • Step 3 Ethyl 10'-(difluoromethyl)-9'-(3-hvdroxy-3-methylbutoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 3 10'-(Difluoromethvn-9'-(3-hvdroxy-3-methylbutoxyV2'-oxo-2'.7'- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • Step 1 Oxetan-3-ylmethyl 4-methylbenzenesulfonate
  • Step 2 Ethyl 10'-formyl-9'-hvdroxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane-
  • Step 3 Ethyl 10'-formyl-9'-(oxetan-3-ylmethoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 4 Ethyl 10'-(difluoromethyl)-9'-(oxetan-3-ylmethoxy)-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 5 10'-(Difluoromethyl)-9'-(oxetan-3-ylmethoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • Step 1 3-Methoxy-3-methylbutyl 4-methylbenzenesulfonate
  • Step 2 Ethyl 10'-formyl-9'-(3-methoxy-3-methylbutoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 3 Ethyl 10'-(difluoromethyl)-9'-(3-methoxy-3-methylbutoxy)-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 4 10'-(DifluoromethylV9'-(3-methoxy-3-methylbutoxyV2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • Step 1 (3-Methyloxetan-3-yl)methyl 4-methylbenzenesulfonate
  • Step 2 Ethyl 10'-formyl-9'-((3-methyloxetan-3-yl)methoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 3 Ethyl 10'-(difluoromethyl)-9'-((3-methyloxetan-3-yl)methoxy)-2'-oxo- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
  • Step 4 10'-(Difluoromethvn-9'-((3-methyloxetan-3-vnmethoxyV2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
  • Step 1 4-Methoxy-3-(3-methoxypropoxy)benzaldehvde
  • Step 3 -(Bromomethyl)-1 -methoxy-2-(3-methoxypropoxy)benzene

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Abstract

Compounds, specifically hepatitis B virus and/or hepatitis D virus inhibitors, more specifically compounds that inhibit HBe antigen and HBs antigen in a subject, for the treatment of viral infections, and methods of preparing and using such compounds. Formula (I):

Description

DIHYDROQUINOLIZINONES AS ANTIVIRALS
FIELD OF THE INVENTION
[0001] The present invention relates to compounds useful for treatment of HBV in animals, and more particularly for treatment of HBV in humans.
BACKGROUND OF THE INVENTION
[0002] Hepatitis B is a viral disease transmitted parenterally by contaminated material such as blood and blood products, contaminated needles, sexually and vertically from infected or carrier mothers to their offspring. In those areas of the world where the disease is common vertical transmission at an early age results in a high proportion of infected individuals becoming chronic carriers of hepatitis B. It is estimated by the World Health Organization that more than 2 billion people have been infected worldwide, with about 4 million acute cases per year, 1 million deaths per year, and 350-400 million chronic carriers. Approximately 25% of carriers die from chronic hepatitis, cirrhosis, or liver cancer and nearly 75% of chronic carriers are Asian. Hepatitis B virus (HBV) is the second most significant carcinogen behind tobacco, causing from 60% to 80% of all primary liver cancer. HBV is 100 times more contagious than HIV.
[0003] HBV is transmitted through percutaneous or parenteral contact with infected blood, body fluids, and by sexual intercourse. HBV is able to remain on any surface it comes into contact with for about a week, e.g. table-tops, razor blades, blood stains, without losing infectivity.
However, HBV cannot cross the skin or the mucous membrane barrier. Some break in this barrier, which can be minimal and insignificant, is required for transmission.
[0004] HBV is a small enveloped DNA virus belonging to the hepadnavirus family. The virus replicates through an RNA intermediate form by reverse transcription, which in practice relates them to retroviruses, like HIV. Although replication takes place in the liver, the virus spreads to the blood where viral proteins and antibodies against them are found in infected people. HBV is many times more infectious than HIV due to the greater concentrations of HBV virus found in the bloodstream at any given time.
[0005] HBV infection results in the production of two different particles: 1 ) the HBV virus itself (or Dane particle) which includes a viral capsid assembled from the HBV core antigen protein (HBcAg) and is covered by the hepatitis B surface antigen (HBsAg) and is capable of reinfecting cells and 2) subviral particles (or SVPs) which are high density lipoprotein-like particles comprised of lipids, cholesterol, cholesterol esters and the small and medium forms of the hepatitis B surface antigen (HBsAg) which are non-infectious. For each viral particle produced, 1 ,000-10,000 SVPs are released into the blood. As such SVPs (and the HBsAg protein they carry) represent the
l overwhelming majority of viral protein in the blood. HBV infected cells also secrete a soluble proteolytic product of the pre-core protein called the HBV e-antigen (HBeAg).
[0006] The hepatitis D virus (HDV) uses HBsAg to form its viral structure (Taylor, 2006, Virology, 344: 71 -76) and as such, HDV infection can only occur in subjects with concomitant HBV infection. While the incidence of HDV co-infection in asymptomatic HBV carriers and chronic HBV- related liver disease is low in countries with a low incidence of HBV infection, it is a significant complication in HBV-infected subjects in countries with a high incidence of HBV infection and can increase the rate of progression of liver disease to fulminant hepatitis. As such, the clear unmet medical need in HBV infection is even more pressing in HBV/HDV co-infected subjects.
[0007] The current conventional methods of treatment for HBV include interferon or thymosin a1 -based immunotherapies and the suppression of viral production by inhibition of the HBV polymerase (e.g., "nucs"). HBV polymerase inhibitors are effective in reducing viral production, but have little to no effect in rapidly reducing HBsAg blood levels or can slowly reduce HBsAg with long term treatment in a limited number of patients (as is the case with tenofovir disoproxil fumarate). Interferon based immunotherapy can achieve a reduction of both viral production and early removal of HBsAg from the blood but only in a small percentage of treated subjects. The generally accepted role of HBsAg in the blood is to sequester anti-HBsAg antibodies and allow infectious viral particles to escape immune detection which is likely one of the reasons why HBV infection remains a chronic condition. In addition HBsAg, HBeAg and HBcAg all have immuno-inhibitory properties as discussed below and the persistence of these viral proteins in the blood of patients following the administration of any of the currently available treatments for HBV as described above is likely having a significant impact in preventing patients from achieving immunological control of their HBV infection.
[0008] Although the three primary HBV proteins (HBsAg, HBeAg and HBcAg) all have immunoinhibitory properties (see below), HBsAg comprises the overwhelming majority of HBV protein in the circulation of HBV infected subjects. Additionally, while the removal (via
seroconversion) of HBeAg or reductions in serum viremia are not correlated with the development of sustained control of HBV infection off treatment, the removal of serum HBsAg from the blood (and seroconversion) in HBV infection is a well-recognized excellent prognostic indicator of antiviral response on treatment which will lead to control of HBV infection off treatment (although this only occurs in a small fraction of patients receiving immunotherapy). Thus, while reduction of all three major HBV proteins (HBsAg, HBeAg and HBcAg) may result in the optimal removal of inhibitory effect, the removal of HBsAg alone may be sufficient in and of itself to remove the bulk of the viral inhibition of immune function in subjects with HBV infection.
[0009] Therefore, in the absence of any current treatment regimen which can restore immunological control of HBV in a large proportion of patients, there is a need to be provided with an effective treatment against HBV infection and HBV/HDV co-infection which can restore immunological control in the majority of patients.
[0010] Hepatitis B viral infections, in conjunction with Hepatitis D viral infections, are a continuing medical problem because, like any rapidly-replicating infectious agent, there are continuing mutations that help some sub-populations of HBV become resistant to current treatment regimens. At the present time there are no effective therapeutic agents for treating humans infected with HBV and/or Hepatitis D virus (HDV) infections which result in seroconversion to the virus in the body, or which effect a 90% reduction of antigen, compared to baseline numbers before treatment, in persons suffering from a hepatitis B viral infection. Currently the
recommended therapies for chronic HBV and/or HDV infection by the American Association for the Study of Liver Diseases (AASLD) and the European Association for the Study of the Liver (EASL) include interferon alpha (INFa), pegylated interferon alpha-2a (Peg-IFN2a), entecavir, and tenofovir. However, typical interferon therapy is 48-weeks and results in serious and unpleasant side effects, and HBeAg seroconversion, 24 weeks after therapy has ceased, ranges from only 27- 36%. Seroconversion of HBsAg is even lower - only 3% observed immediately after treatment ceases, with an increase to upwards of 12% after 5 years.
[0011] The nucleoside and nucleotide therapies entecavir and tenofovir are successful at reducing viral load, but the rates of HBeAg seroconversion and HBsAg loss are even lower than those obtained using IFNa therapy. Other similar therapies, including lamivudine (3TC), telbivudine (LdT), and adefovir are also used, but for nucleoside/nucleotide therapies in general, the emergence of resistance limits therapeutic efficacy.
[0012] Recent clinical research has found a correlation between seroconversion and reductions in HBeAg (Fried et al (2008) Hepatology 47:428) and reductions in HBsAg (Moucari et al (2009) Hepatology 49:1 151). Reductions in antigen levels may have allowed immunological control of HBV infection because high levels of antigens are thought to induce immunological tolerance. Current nucleoside therapies for HBV are capable of dramatic reductions in serum levels of HBV but have little impact on HBeAg and HBsAg levels. Antisense therapy differs from nucleoside therapy in that it can directly target the transcripts for the antigens and thereby reduce serum HBeAg and HBsAg levels. But antisense therapy is expensive and requires delivery via injection.
[0013] Thus, there is a need in the art to discover and develop new anti-viral therapies. More particularly, there is a need for new anti-HBV therapies capable of increasing HBeAg and HBsAg seroconversion rates. These serum markers are indicative of immunological control of HBV infection and leads to an improved prognosis, i.e. prevention of liver disease and progression to cirrhosis, prevention of liver failure, prevention of hepatocellular cancer (HCC), prevention of liver disease-related transplantation, and prevention of death. SUMMARY OF THE INVENTION
[0014] An embodiment of the present invention features a compound of Formula I:
Figure imgf000005_0001
Formula I wherein
[0015] R1 and R4 are independently selected from hydrogen; halogen; hydroxy; alkoxy; amino; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl;
unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci-6alkyoxy; unsubstituted C3.
ycycloalkyl or halo- or alkyl-substituted C3-7cycloalkyl; unsubstituted C3-7cycloalkenyl or halo- or alkyl-substituted C3-7cycloalkenyl; unsubstituted -CxH2x-phenyl or halo- or alkyl- substituted -CxH2x-phenyl; unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted - O-Cxhbx-phenyl; and unsubstituted -(Ci_6alkyl)N- CxH2x-phenyl or halo- or alkyl-substituted -(Ci-6alkyl)N- CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[0016] R2 and R3 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR20 R21 ; COR19; COOR19; CONR20 R21 ; pyrrolidinyl; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; Ci-6alkylCOOR19; -BOR19OR19'; unsubstituted Ci- 6alkoxy or halo- or alkyl-substituted Ci-6alkyoxy; unsubstituted C3.7cycloalkyl or halo- or alkyl-substituted C3.7cycloalkyl; unsubstituted C3.7cycloalkenyl or halo- or alkyl-substituted C3.7cycloalkenyl; unsubstituted or halo- or alkyl-substituted monocyclic heterocycloalkyi ring wherein a heteroatom in the heterocycloalkyi ring is O, N or S; unsubstituted or halo- or alkyl-substituted heteroaryl ring wherein a heteroatom in the heteroaryl ring is O, N or S; pyrrolidinyl; furanyl; pyrrolyl; imidazolyl; unsubstituted -CxH2x-phenyl or halo- or alkyl- substituted -CxH2X-phenyl; unsubstituted -0-CxH2X-phenyl or halo- or alkyl-substituted -O- CxH2x-phenyl; unsubstituted -(Ci-6alkyl)N- CxH2x-phenyl or halo- or alkyl-substituted -(Ci_ 6alkyl)N- CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;and -OR12; or R2 and R3 together form a 3 to 8 membered cycloalkyl ring; or R2 and R3 together form a 3 to 8 membered heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O, N, NR22, S, SR22 or SR22R22'; or R2 and R3 together form a 3 to 8 member aryl ring; or R2 and R3 together form a 3 to 8 member heteroaryl ring, wherein a heteroatom in the heteroaryl ring is O, N, NR22, S, SR22 or SR22R22'; [0017] R5 and R6 are independently selected from halogen; cyano; unsubstituted Ci-6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3.
ecycloalkyl; unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl;
unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl and heteroaryl; or R5 and R6 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R13, R13', R14 and/or R14', or R5 and R6 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R13, R13', R14 and/or R14', wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
[0018] R7 and R8 are selected from independently hydrogen; halogen; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3-8cycloalkyl; unsubstituted CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R7 and R8 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18', or R7 and R8 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R17, R17', R18 and/or R18', wherein a heteroatom in the heterocycloalkyi ring is O, N, NR22, S, SR22 or SR22R22';
[0019] R9 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3.
ecycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted or halo- or alkyl substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[0020] R10 is selected from COOR19 and CONR20 R216;
[0021] R11 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkyoxy; unsubstituted or halo- or alkyl-substituted C3.7cycloalkyl;
unsubstituted CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[0022] R12 is selected from hydrogen; unsubstituted Ci_6alkyl or Ci_6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, heterocycloalkyi, heteroaryl, C3.8cycloalkyl and C2.6alkenyl; Ci-6alkylCi-6alkoxy; Ci_6alkylCi- 6alkoxyCi-6alkoxy; Ci.8alkylamino; Ci-8alkylaminocarbonylCi-6alkyl; Ci-6alkylaminosulfonylCi- 6alkyl; Ci_6alkylsulfonylCi-6alkyl; Ci-6alkylcyano; Ci-6alkylC3-8cycloalkyl; Ci-6alkylC3- ecycloalkylcyano; Ci-6alkylphenyl; Ci-6alkylcarbonylpyrrolidinyl; C2-6alkynyl; C2.6alkynylCi- 6hydroxy; Ci-6alkylCi-6alkoxyamino; Ci-6alkylCi-6alkoxyaminoCi-6alkyl; Ci_6alkylCi- 6alkoxyaminodiCi-6alkyl; Ci-6alkylcarboxy; Ci-6alkylaminocarbonylCi-6alkoxy; Ci_
6alkylheteroaryl, wherein the heteroaryl is a monocyclic heteroaryl comprising O and/or ; and Ci-6alkylheterocycloalkyl, wherein the heterocycloalkyi is a monocyclic heterocycloalkyi comprising O and/or N;
[0023] R13 , R13', R14 and R14' are independently selected from hydrogen,; halogen; amino; aminoalkyl; cyano; Ci_6alkyl; Ci-6alkoxy; carbonyl; carboxamide; and amide; or R13 and R13' or R14 and R14' together form a 3- to 8-membered cycloalkyl; or R14 and R14' together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
[0024] R17 , R17', R18 and R18' are independently selected from hydrogen; halogen; amino; cyano; Ci_6alkyl; Ci-6alkoxy; carbonyl; carboxamide; and amide; or R17 and R18 or R17' and R18' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci-6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
[0025] R19, R19' and R19" are independently selected from hydrogen; Ci_6alkyl; C3. ecycloalkyl; C2.6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci_
6alkyltetrazole; Ci-6alkylthiazole; Ci-6alkyloxazole; Ci-6alkyldioxazole; and Ci_
6alkyloxazolidone; and
[0026] R20 and R21 are independently selected from hydrogen; Ci_6alkyl; C3.
ecycloalkyl; C2.6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci_
6alkyltetrazole; Ci-6alkylthiazole; Ci-6alkyloxazole; Ci-6alkyldioxazole; and Ci_
6alkyloxazolidone; or R20 and R21 together with the nitrogen to which they are attached form unsubstituted pyrrolidinyl, unsubstituted piperidinyl, or unsubstituted morpholinyl; or R20 and R21 together with the nitrogen to which they are attached form carboxyl-substituted pyrrolidinyl, carboxyl-substituted piperidinyl or carboxyl-substituted morpholinyl;
[0027] R22 and R22' are independently selected from hydrogen; oxygen; Ci_6alkyl; Ci-6haloalkyl; C3.8cycloalkyl; Ci-6alkylC3-8cycloalkyl; C2.6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci-6alkyltetrazole; Ci-6alkylthiazole; Ci-6alkyloxazole; Ci_ ealkyldioxazole; Ci-6alkyloxazolidone; -COR19; -COOR19'; -CSOR19 "; and -CONR20R21 ; and [0028] R23 is selected from hydrogen, Ci_6alkyl; C3-8cycloalkyl; C2-6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci-6alkyltetrazole; Ci-6alkylthiazole; Ci_ 6alkyloxazole; Ci.6alkyldioxazole; Ci.6alkyloxazolidone; aryl; heteroaryl; and benzyl, or a pharmaceutically acceptable salt thereof.
[0029] One embodiment of the invention provides a compound of Formula I
Figure imgf000008_0001
Formula I wherein
[0030] R1 and R4 are independently selected from hydrogen; halogen; hydroxy; alkoxy; amino; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl;
unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci-6alkyoxy; unsubstituted C3.
ycycloalkyl or halo- or alkyl-substituted C3-7cycloalkyl; unsubstituted C3-7cycloalkenyl or halo- or alkyl-substituted C3.7cycloalkenyl; unsubstituted -CxH2x-phenyl or halo- or alkyl- substituted -CxH2x-phenyl; unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -O- CxH2x-phenyl; and unsubstituted -(Ci-6alkyl)N- CxH2x-phenyl or halo- or alkyl-substituted - (Ci-6alkyl)N- CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[0031] R2 and R3 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR20 R21 ; COR19; COOR19; CONR20 R21 ; pyrrolidinyl; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; Ci-6alkylCOOR19; -BOR19OR19'; unsubstituted Ci- 6alkoxy or halo- or alkyl-substituted Ci-6alkyoxy; unsubstituted C3.7cycloalkyl or halo- or alkyl-substituted C3.7cycloalkyl; C3.7cycloalkenyl or halo- or alkyl-substituted C3.
7cycloalkenyl; unsubstituted or halo- or alkyl-substituted monocyclic heterocycloalkyi ring wherein a heteroatom in the heterocycloalkyi ring is O, N or S; unsubstituted or halo- or alkyl-substituted heteroaryl ring wherein a heteroatom in the heteroaryl ring is O, N or S, pyrrolidinyl, furanyl, pyrrolyl, imidazolyl, substituted -CxH2x-phenyl or unsubstituted -CxH2x- phenyl, substituted -0-CxH2x-phenyl or unsubstituted -0-CxH2x-phenyl, substituted -(Ci_ 6alkyl)N- CxH2x-phenyl or unsubstituted -(Ci_6alkyl)N- CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; and -OR12; or R2 and R3 together form a 3 to 8 membered cycloalkyl ring; or R2 and R3 together form a 3 to 8 membered heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O, N, NR22, S, SR22 or SR22R22'; or R2 and R3 together form a 3 to 8 member aryl ring; or R2 and R3 together form a 3 to 8 member heteroaryl ring, wherein a heteroatom in the heteroaryl ring is O, N, NR22, S, SR22 or SR22R22';
[0032] R5 is selected from hydrogen and unsubstituted Ci_6alkyl or halo- or alkyl- substituted Ci_6alkyl;
[0033] R6 and R7 together form a 3- to 8-membere cycloalkyl ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R15, R15', R16 and/or R16'; or R6 and R7 together form a 3- to 8- membered heterocycloalkyi ring comprising one heteroatom or two or more heteroatoms, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R15, R15', R16 and/or R16', wherein the one heteroatom in the heterocycloalkyi ring is NR20 and the two or more heteroatoms are selected from N, NR22, O, S, SR22 and SR22R22';
[0034] R8 is selected from hydrogen and unsubstituted Ci_6alkyl or halo- or alkyl- substituted Ci_6alkyl;
[0035] R9 is selected from hydrogen, hydroxy, halogen, cyano, unsubstituted Ci_ 6alkyl or halo- or alkyl-substituted Ci_6alkyl, unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkoxy, unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3.
ecycloalkyl, unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl an unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[0036] R10 is selected from COOR19 and CONR20 R21 ;
[0037] R11 is selected from hydrogen, hydroxy, halogen, cyano, unsubstituted Ci_ 6alkyl or halo- or alkyl-substituted Ci_6alkyl, unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkyoxy; unsubstituted C3-7cycloalkyl or halo- or alkyl substituted C3.
ycycloalkyl; unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[0038] R12 is hydrogen; unsubstituted Ci_6alkyl or Ci_6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, heterocycloalkyi, heteroaryl, C3.8cycloalkyl and C2.6alkenyl; Ci-6alkylCi-6alkoxy; Ci-6alkylCi-6alkoxyCi-6alkoxy; Ci-8alkylamino; Ci-8alkylaminocarbonylCi-6alkyl; Ci-6alkylaminosulfonylCi-6alkyl; Ci_
6alkylsulfonylCi-6alkyl; Ci-6alkylcyano; Ci-6alkylC3-8cycloalkyl; Ci-6alkylC3-8cycloalkylcyano; Ci-6alkylphenyl; Ci-6alkylcarbonylpyrrolidinyl; C2.6alkynyl; C2-6alkynylCi-6hydroxy; Ci_6alkylCi- 6alkoxyamino; Ci-6alkylCi-6alkoxyaminoCi-6alkyl; Ci-6alkylCi-6alkoxyaminodiCi-6alkyl; Ci_ 6alkylcarboxy; Ci-6alkylaminocarbonylCi-6alkoxy; Ci-6alkylheteroaryl, wherein the heteroaryl is an O-containing or N-containing monocyclic heteroaryl; or Ci-6alkylheterocycloalkyl, wherein the heterocycloalkyi is monocyclic heterocycloalkyi; [0039] R15, R15', R16 and R16' are independently hydrogen, halogen, amino, cyano, Ci-6alkyl, or Ci-6alkoxy, wherein any or R15, R15', R16 or R16', together with R5 or R8, form a 3- to 8-membered cycloalkyl ring or a 3-8 membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2.
6alkenyl or Ci_6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22'; or any of R15, R15', R16 and/or R16' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
[0040] R19, R19' and R19"are independently hydrogen, Ci_6alkyl, C3-8cycloalkyl, C2. 6alkenyl, phenyl, Ci-6alkylimidizole, Ci-6alkoxy, Ci-6alkyltriazole, Ci-6alkyltetrazole, Ci_
6alkylthiazole, Ci-6alkyloxazole, Ci-6alkyldioxazole; Ci-6alkyloxazolidone;
[0041] R20 and R21 are independently hydrogen, Ci_6alkyl, C3-8cycloalkyl, C2.
6alkenyl, phenyl, Ci-6alkylimidizole, Ci-6alkoxy, Ci-6alkyltriazole,Ci-6alkyltetrazole, Ci_
6alkylthiazole, Ci-6alkyloxazole, Ci-6alkyldioxazole; Ci-6alkyloxazolidone, -CONR22R22'; or R20 and R21 together with the nitrogen to which they are attached form unsubstituted pyrrolidinyl, unsubstituted piperidinyl, or unsubstituted morpholinyl; or form carboxyl-substituted pyrrolidinyl, carboxyl-substituted piperidinyl or carboxyl-substituted morpholinyl;
[0042] R22 and R22' are independently selected from hydrogen, oxygen, Ci_6alkyl, Ci-6haloalkyl, C3-8cycloalkyl, Ci-6alkylC3-8cycloalkyl, C2.6alkenyl, phenyl, Ci-6alkylimidizole, Ci-6alkoxy, Ci-6alkyltriazole,Ci-6alkyltetrazole, Ci-6alkylthiazole, Ci-6alkyloxazole, Ci_ ealkyldioxazole; Ci-6alkyloxazolidone, -COR19, -COOR19', -CSOR19 ", -CONR20R21 ; and
[0043] R23 is hydrogen, Ci_6alkyl, C3.8cycloalkyl, C2.6alkenyl, phenyl, Ci_
6alkylimidizole, Ci.6alkoxy, Ci-6alkyltriazole, Ci.6alkyltetrazole, Ci.6alkylthiazole, Ci_
6alkyloxazole, Ci.6alkyldioxazole; Ci.6alkyloxazolidone; aryl, heteroaryl or benzyl, or a pharmaceutically acceptable salt thereof.
[0044] One embodiment of the invention provides a compound of Formula I
Figure imgf000010_0001
Formula I wherein [0045] R1, and R4 are independently selected from hydrogen; halogen; hydroxy; alkoxy; amino; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl;
unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci-6alkyoxy; unsubstituted C3.
ycycloalkyl or halo- or alkyl-substituted C3-7cycloalkyl; unsubstituted C3-7cycloalkenyl or halo- or alkyl-substituted C3-7cycloalkenyl; unsubstituted -CxH2x-phenyl or halo- or alkyl- substituted -CxH2x-phenyl; unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -O- CxH2x-phenyl; and unsubstituted -(Ci-6alkyl)N- CxH2x-phenyl or halo- or alkyl-substituted - (Ci-6alkyl)N- CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[0046] R2 and R3 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR20 R21 ; COR19; COOR19; CONR20 R21 ; pyrrolidinyl; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; Ci-6alkylCOOR19'; -BOR19OR19'; unsubstituted Ci- 6alkoxy or halo- or alkyl-substituted Ci-6alkyoxy; C3-7cycloalkyl or halo- or alkyl-substituted C3-7cycloalkyl; unsubstituted C3.7cycloalkenyl or halo- or alkyl-substituted C3.7cycloalkenyl; unsubstituted or halo- or alkyl-substituted monocyclic heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O, N or S; unsubstituted or halo- or alkyl- substituted heteroaryl ring wherein a heteroatom in the heteroaryl ring is O, N or S;
pyrrolidinyl; furanyl; pyrrolyl; imidazolyl; unsubstituted -CxH2x-phenyl or halo- or alkyl- substituted -CxH2x-phenyl; unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted - 0-CxH2x-phenyl; unsubstituted -(Ci-6alkyl)N- CxH2x-phenyl or halo- or alkyl-substituted - (Ci-6alkyl)N- CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; and -OR12; or R2 and R3 together form a 3 to 8 member cycloalkyi ring; or R2 and R3 together form a 3 to 8 member heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O, N, NR22, S, SR22 or SR22R22'; or R2 and R3 together form a 3 to 8 member aryl ring; or R2 and R3 together form a 3 to 8 member heteroaryl ring, wherein a heteroatom in the heteroaryl ring is O, N, NR22, S, SR22 or SR22R22';
[0047] R5 and R6 are independently selected from hydrogen; halogen; cyano; unsubstituted Ci_6alkyl or halo-or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_6alkoxy; unsubstituted C3-ecycloalkyl or halo- or alkyl-substituted C3. ecycloalkyl; unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R5 and R6 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R13, R13', R14 and/or R14', or R5 and R6 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R13, R13', R14 and/or R14', wherein a heteroatom in the heterocycloalkyi ring is O, N, NR22, S, SR22 or SR22R22'; [0048] R7 and R8 are independently selected from halogen; cyano; unsubstituted Ci-6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3- ecycloalkyl; unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl;
unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; and heteroaryl; or R7 and R8 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18', or R7 and R8 together form a 3- to 8- membered
heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R17 , R18', R17 and/or R18', wherein a heteroatom in the heterocycloalkyi ring is O, N, NR22, S, SR22 or SR22R22';
[0049] R9 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6alkyl or halo-or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3-ecycloalkyl or halo- or alkyl-substituted C3- ecycloalkyl; unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[0050] R10 is selected from COOR19and CONR20 R21;
[0051] R11 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkyoxy; unsubstituted or halo- or alkyl-substituted C3-7cycloalkyl;
unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[0052] R12 is selected from hydrogen; unsubstituted Ci_6alkyl or Ci_6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, C3- ecycloalkyl and C2.6alkenyl; Ci-6alkylCi-6alkoxy; Ci-6alkylCi-6alkoxyCi-6alkoxy; Ci-8alkylamino; Ci-8alkylaminocarbonylCi-6alkyl; Ci-6alkylaminosulfonylCi-6alkyl; Ci-6alkylsulfonylCi_6alkyl; Ci-6alkylcyano; Ci-6alkylC3-8cycloalkyl; Ci-6alkylC3-8cycloalkylcyano; Ci-6alkylphenyl; Ci- 6alkylcarbonylpyrrolidinyl; C2.6alkynyl; C2-6alkynylCi-6hydroxy; Ci-6alkylCi-6alkoxyamino; Ci_ 6alkylCi-6alkoxyaminoCi-6alkyl; Ci-6alkylCi-6alkoxyaminodiCi-6alkyl; Ci-6alkylcarboxy; Ci_ 6alkylaminocarbonylCi-6alkoxy; Ci-6alkylheteroaryl, wherein the heteroaryl is a monocyclic heteroaryl comprising O and/or ; and Ci-6alkylheterocycloalkyl, wherein the heterocycloalkyi is a monocyclic heterocycloalkyi comprising O and/or N;
[0053] R13 , R13', R14 and R14' are independently selected from hydrogen; halogen; amino; aminoalkyl; cyano; Ci_6alkyl; Ci-6alkoxy; carbonyl; carboxamide; and amide; or R13 and R14 or R13' and R14' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
[0054] R17, R17', R18 and R18' are independently selected from hydrogen; halogen; amino; cyano; Ci_6alkyl; and Ci-6alkoxy; or R17 and R18 or R17' and R18' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2. 6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
[0055] R19, R19' and R19" are independently selected from hydrogen; Ci-ealkyl; C3. ecycloalkyl; C2-6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci_
6alkyltetrazole; Ci-6alkylthiazole; Ci-6alkyloxazole; Ci-6alkyldioxazole; and Ci_
6alkyloxazolidone; and
[0056] R20 and R21 are independently selected from hydrogen; Ci_6alkyl; C3. ecycloalkyl; C2-6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci- 6alkyltetrazole; Ci-6alkylthiazole; Ci-6alkyloxazole; Ci-6alkyldioxazole; and Ci_
6alkyloxazolidone; or R20 and R21 together with the nitrogen to which they are attached form unsubstituted pyrrolidinyl, unsubstituted piperidinyl, or unsubstituted morpholinyl; or form carboxyl-substituted pyrrolidinyl, carboxyl-substituted piperidinyl or carboxyl-substituted morpholinyl; and
[0057] R22 and R22' are independently selected from hydrogen; oxygen; Ci_6alkyl; Ci-6haloalkyl, C3-8cycloalkyl; Ci-6alkylC3-8cycloalkyl; C2-6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci-6alkyltetrazole; Ci-6alkylthiazole; Ci-6alkyloxazole; Ci_ ealkyldioxazole; Ci-6alkyloxazolidone; -COR19; -COOR19'; -CSOR19 "; and -CONR20R21 ; and
[0058] R23 is selected from hydrogen; Ci_6alkyl; C3.8cycloalkyl; C2-6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci-6alkyltetrazole; Ci-6alkylthiazole; Ci_
6alkyloxazole; Ci.6alkyldioxazole; Ci.6alkyloxazolidone; aryl; heteroaryl and benzyl, or a pharmaceutically acceptable salt thereof.
[0059] Another embodiment provides a compound of Formula IA or Formula IB:
Figure imgf000014_0001
Formula I Formula IA Formula IB
[0060] wherein C* is a carbon atom stereocenter which has a configuration which is (R) or (S);
[0061] R1, R2, R3 and R4 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR20 R21 COR19; COOR19; CONR20 R21 ; pyrrolidinyl; unsubstituted Ci-6alkyl or halo- or alkyl substituted Ci_6alkyl; Ci-6alkylCOOR19; -BOR19OR19'; unsubstituted Ci-6alkoxy or halo-or alkyl-substituted Ci-6alkyoxy; unsubstituted C3-7cycloalkyl or halo-or alkyl-substituted C3-7cycloalkyl; unsubstituted C3-7cycloalkenyl or halo- or alkyl-substituted C3-7cycloalkenyl; unsubstituted or halo- or alkyl-substituted monocyclic heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O, N or S; unsubstituted or halo- or alkyl-substituted heteroaryl ring wherein a heteroatom in the heteroaryl ring is O, N or S; pyrrolidinyl; furanyl; pyrrolyl; imidazolyl; unsubstituted or halo- or alkyl-substituted -CxH2x- phenyl; unsubstituted or halo- or alkyl-substituted -O-Cxhbx-phenyl; unsubstituted or halo- or alkyl-substituted -(Ci-6alkyl)N-
Figure imgf000014_0002
wherein x is 0, 1 , 2, 3, 4, 5 or 6; and - OR12; or R2 and R3 together form a 3 to 8 member cycloalkyl ring; or R2 and R3 together form a 3 to 8 member heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O, N, NR22, S, SR22 or SR22R22'; or R2 and R3 together form a 3 to 8 member aryl ring; or R2 and R3 together form a 3 to 8 member heteroaryl ring, wherein a heteroatom in the heteroaryl ring is O, N, NR22, S, SR22 or SR22R22';
[0062] and
[0063] (A) R5 and R6 are independently selected from halogen; cyano;
unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci-6alkoxy; unsubstituted C3.8cycloalkyl or halo- or alkyl-substituted C3.8cycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl; unsubstituted or halo- or alkyl-substituted -O-Cxhbx-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; and heteroaryl; or R5 and R6 together form a 3- to 8-membered cycloalkyl ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R13, R13', R14 and/or R14', or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R13, R13', R14 and/or R14', wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; and
[0064] R7 and R8 are independently hydrogen; halogen; cyano; unsubstituted Ci_ 6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3-ecycloalkyl or halo- or alkyl-substituted C3- ecycloalkyl; unsubstituted or halo- or alkyl-substituted-CxH2x-phenyl; unsubstituted or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R7 and R8 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18', or R7 and R8 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R17, R17', R18 and/or R18', wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; or
(B) R5 and R8 are independently selected from hydrogen, unsubstituted Ci_6alkyl, and halo- or alkyl-substituted Ci_6alkyl; and
[0065] R6 and R7 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R15, R15', R16 and/or R16'; or R6 and R7 together form a 3- to 8- membered heterocycloalkyi ring comprising one heteroatom or two or more heteroatoms, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R15, R15', R16 and/or R16', wherein the one heteroatom in the heterocycloalkyi ring is NR20 and the two or more heteroatoms are selected from N, NR22, O, S, SR22 and SR22R22'; or
[0066] (C) R5 and R6 are independently selected from hydrogen; halogen; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci-6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl- substituted C3-8cycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl;
unsubstituted or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R5 and R6 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18', or R5 and R6 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R13 , R13', R14 and/or R14', wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; and
[0067] R7 and R8 are independently selected from halogen; cyano; unsubstituted Ci-6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3-ecycloalkyl or halo- or alkyl-substituted C3- ecycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl; unsubstituted or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; and heteroaryl; or R7 and R8 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18', or R7 and R8 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R17 , R18', R17 and/or R18', wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; and
[0068] wherein
[0069] R9 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3.
ecycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted or halo- or alkyl-substituted-0-CxH2)^phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[0070] R10 is selected from COOR19and CONR20 R21;
[0071] R11 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkyoxy; unsubstituted or halo- or alkyl-substituted C3-7cycloalkyl;
unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[0072] R12 is selected from hydrogen; unsubstituted Ci_6alkyl or Ci_6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, heterocycloalkyi, heteroaryl, C3-8cycloalkyl and C2-6alkenyl; Ci-6alkylCi-6alkoxy; Ci_6alkylCi- 6alkoxyCi-6alkoxy; Ci.8alkylamino; Ci-8alkylaminocarbonylCi-6alkyl; Ci-6alkylaminosulfonylCi- 6alkyl; Ci_6alkylsulfonylCi-6alkyl; Ci-6alkylcyano; Ci-6alkylC3-8cycloalkyl; Ci-6alkylC3- ecycloalkylcyano; Ci-6alkylphenyl; Ci-6alkylcarbonylpyrrolidinyl; C2.6alkynyl; C2.6alkynylCi- 6hydroxy; Ci-6alkylCi-6alkoxyamino; Ci-6alkylCi-6alkoxyaminoCi-6alkyl; Ci_6alkylCi- 6alkoxyaminodiCi-6alkyl; Ci-6alkylcarboxy; Ci-6alkylaminocarbonylCi-6alkoxy; Ci_
6alkylheteroaryl, wherein the heteroaryl is an O-containing or N-containing monocyclic heteroaryl; or Ci-6alkylheterocycloalkyl, wherein the heterocycloalkyi is a monocyclic heterocycloalkyi;
[0073] R13 , R13', R14 and R14' are independently selected from hydrogen; halogen; amino; aminoalkyl; cyano; Ci_6alkyl; Ci-6alkoxy; carbonyl; carboxamide; and amide; or R13 and R13' or R14 and R14' together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-ecycloalkyl, C2-6alkenyl or Ci_6alkoxy, wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; [0074] R15, R15', R16 and R16' are independently hydrogen, halogen, amino, cyano, Ci-6alkyl, or Ci-6alkoxy, wherein any or R15, R15', R16 or R16', together with R5 or R8, form a 3- to 8-membered cycloalkyl ring or a 3-8 membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2.
6alkenyl or Ci_6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22'; or any of R15, R15', R16 and/or R16' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
[0075] R17 , R17', R18 and R18' are independently selected from hydrogen; halogen; amino; cyano; Ci_6alkyl; and Ci-6alkoxy; or R17 and R18 or R17' and R18' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2.
6alkenyl or Ci_6alkoxy, wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22';
[0076] R19, R19' and R19" are independently selected from hydrogen; Ci-6alkyl; C3. ecycloalkyl; C2-6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci_
6alkyltetrazole; Ci-6alkylthiazole; Ci-6alkyloxazole; Ci-6alkyldioxazole; and Ci_
6alkyloxazolidone; and
[0077] R20 and R21 are independently selected from hydrogen; Ci_6alkyl; C3.
ecycloalkyl; C2.6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci_
6alkyltetrazole; Ci-6alkylthiazole; Ci-6alkyloxazole; Ci-6alkyldioxazole; and Ci_
6alkyloxazolidone; or R20 and R21 together with the nitrogen to which they are attached form unsubstituted pyrrolidinyl, unsubstituted piperidinyl, or unsubstituted morpholinyl; or form carboxyl-substituted pyrrolidinyl, carboxyl-substituted piperidinyl or carboxyl-substituted morpholinyl; and
[0078] R22 and R22' are independently selected from hydrogen; oxygen; Ci_6alkyl; Ci-6haloalkyl; C3.8cycloalkyl; Ci-6alkylC3-8cycloalkyl; C2.6alkenyl; phenyl; Ci-6alkylimidizole; Ci_6alkoxy; Ci-6alkyltriazole; Ci-6alkyltetrazole; Ci-6alkylthiazole; Ci-6alkyloxazole; Ci- ealkyldioxazole; Ci-6alkyloxazolidone; -COR19; -COOR19'; -CSOR19 "; and -CONR20R21 ; and
[0079] R23 is selected from hydrogen; Ci_6alkyl; C3.8cycloalkyl; C2.6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci.6alkyltetrazole; Ci.6alkylthiazole; Ci_
6alkyloxazole; Ci.6alkyldioxazole; Ci.6alkyloxazolidone; aryl; heteroaryl; and benzyl, or a pharmaceutically acceptable salt thereof.
[0080] A particular embodiment provides a compound selected from the group consisting of: 9'-(Benzyloxy)-10'-bromo-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(Cyclopropylmethoxy)-10'-(1 ,1 -difluoroethyl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
9'-(Cyclopropylmethoxy)-10'-ethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(Cyclopropylmethoxy)-10'-(difluoromethyl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
9'-(Cyclopropylmethoxy)-10'-(fluoromethyl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
9'-(Cyclopropylmethoxy)-2'-oxo-10'-(trifluoromethyl)-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-Methoxy-9'-(3-methoxypropoxy)-2'-oxo-2,2',3,5,6,7'-hexahydrospiro[pyran-4,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-Chloro-9'-(cyclopropylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(Cyclopropylmethoxy)-10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Methyl-2'-oxo-9'-(pyrrolidin-1 -yl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Methyl-9'-(oxetan-3-ylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(Cyclopentylmethoxy)-10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Methoxy-9'-(1 -methyl-1 H-pyrazol-4-yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
9'-(Furan-3-yl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Chloro-9'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylic acid;
10'-(Difluoromethyl)-2'-oxo-9'-((tetrahydro-2H-pyran-4-yl)methoxy)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-(Difluoromethyl)-2'-oxo-9'-((tetrahydrofuran-3-yl)methoxy)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-(Difluoromethyl)-9'-(3-hydroxy-3-methylbutoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid; 10'-(Difluoromethyl)-9'-(oxetan-3-ylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-(Difluoromethyl)-9'-(3-methoxy-3-methylbutoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-(Difluoromethyl)-9'-((3-methyloxetan-3-yl)methoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
3-Hydroxy-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
3-Fluoro-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-Methoxy-9'-(3-methoxypropoxy)-2',3-dioxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
3,10'-Dimethoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Methyl-2'-oxo-9'-(piperidin-1 -yl)-2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(Cyclopropylmethoxy)-10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Methyl-2'-oxo-9'-(pyrrolidin-1 -yl)-2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(Cyclopropylmethoxy)-10'-(difluoromethyl)-2'-oxo-2',7'-dihydrospiro[cyclohexane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
9- (Benzyloxy)-10-bromo-6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 - a]isoquinoline-3-carboxylic acid;
10- Cyclopropyl-6-isopropyl-6-methyl-9-(1 -methyl-1 H-pyrazol-4-yl)-2-oxo-6,7-dihydro-2H- pyrido[2,1 -a]isoquinoline-3-carboxylic acid;
9-(Cyclopropylmethoxy)-10-(difluoromethyl)-6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H- pyrido[2,1 -a]isoquinoline-3-carboxylic acid;
6-lsopropyl-10-methoxy-9-(3-methoxypropoxy)-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 - a]isoquinoline-3-carboxylic acid;
(S)-6-lsopropyl-10-methoxy-9-(3-methoxypropoxy)-6-methyl-2-oxo-6,7-dihydro-2H- pyrido[2,1 -a]isoquinoline-3-carboxylic acid;
(R)-6-lsopropyl-10-methoxy-9-(3-methoxypropoxy)-6-methyl-2-oxo-6,7-dihydro-2H- pyrido[2,1 -a]isoquinoline-3-carboxylic acid; 9'-(3-Methoxypropoxy)-10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Ethynyl-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(3-Methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3',10'-dicarboxylic acid;
9'-(3-Methoxypropoxy)-10'-(methylcarbamoyl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-Carbamoyl-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Acetyl-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
3-lsopropoxy-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
3,10'-Dimethoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
3-(Cyclopentyloxy)-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-lsopropoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-lsopentyl-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(lsobutylamino)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-((Cyclopropylmethyl)amino)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
9'-Cyano-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylic acid;
10'-(Dimethylamino)-9'-(furan-3-yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(furan-3-yl)-2'-oxo-10'-((1 ,1 ,1 -trifluoro-/V-methylmethyl)sulfonamido)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-(Azetidin-1 -yl)-9'-(furan-3-yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(Furan-3-yl)-10'-(methylamino)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid; 9'-Hexyl-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylic acid;
10'-Methoxy-2'-oxo-9'-propyl-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylic acid;
9'-(Cyclopropylmethoxy)-2'-oxo-10'-propyl-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-Ethyl-10'-methoxy-2'-oxo-2',7',1 1 ',1 1 a'-tetrahydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(isopropoxymethyl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(Cyclopropoxymethyl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(Butoxymethyl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-Butyl-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylic acid;
9'-(3-Ethoxypropyl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Methoxy-2'-oxo-9'-phenethyl-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Bromo-9'-(cyclopropylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(Cyclopropylmethoxy)-10'-(furan-3-yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-Cyclopropyl-9'-(cyclopropylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
1 ',8'-Dibromo-10'-cyclopropyl-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
9'-(Cyclopropylmethoxy)-10'-isopropyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(Cyclopropylmethoxy)-10'-(1 -methyl-1 H-pyrazol-3-yl)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
9'-(Cyclopropylmethoxy)-10'-(3-hydroxypropyl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-(cyclopent-1 -en-1 -yl)-9'-(cyclopropylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid; 10'-Cyclopentyl-9'-(cyclopropylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-Methyl-9'-(1 -methyl-1 /-/-pyrazol-4-yl)-2'-oxo-2',7'-dihydrospiro[cyclohexane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
9'-lsobutoxy-10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(Cyclobutylmethoxy)-10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(2-Cyclopropylethoxy)-10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Methyl-2'-oxo-9'-(pyrrolidin-1 -yl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-lsobutoxy-10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(2-Hydroxy-2-methylpropoxy)-10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
9'-(2-methoxy-2-methylpropoxy)-10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
9'-(Furan-3-yl)-2'-oxo-10'-(pyrrolidin-1 -yl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
3-(benzyloxy)-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-methoxy-2'-oxo-9'-pentyl-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylic acid;
10'-Methoxy-9'-(4-methoxyphenyl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(4-Cyanophenyl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Methoxy-9'-(2-methoxyphenyl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(3-Cyanophenyl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Methoxy-9'-(3-methoxyphenyl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(2-Cyanophenyl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid; 10'-Cethoxy-2'-oxo-9'-(pyridin-4-yl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Methoxy-9'-(2-methoxypyridin-3-yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
9'-(2-Cyclopropylethyl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Methoxy-9'-(6-methoxypyridin-3-yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-Methoxy-9'-(5-methoxypyridin-3-yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-Methoxy-2'-oxo-9'-(pyridin-3-yl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Methoxy-2'-oxo-9'-(1 H-pyrazol-4-yl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(Cyanomethyl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(4-Carboxybutyl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(5-Amino-5-oxopentyl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(4-Cyanobutyl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
2'-Oxo-2',7',10',1 1 '-tetrahydrospiro[cyclobutane-1 ,6'-[1 ,4]dioxino[2,3-g]pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
3,3-Difluoro-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
3-(6-(Diisobutylamino)-5-((3-(trifluoromethyl)-1 ,2,4-thiadiazol-5-yl) amino)pyridin-3- yl)butanoic acid;
10'-Methoxy-9'-(4-methoxybutyl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10-(Difluoromethyl)-9-(furan-3-yl)-6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 - a]isoquinoline-3-carboxylic acid;
10-(Difluoromethyl)-6-isopropyl-9-methoxy-6-methyl-2-oxo-6,7-dihydro-2/-/-pyrido[2,1 - a]isoquinoline-3-carboxylic acid;
10-(Difluoromethyl)-9-ethynyl-6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 - a]isoquinoline-3-carboxylic acid; 10'-(Difluoromethyl)-9'-ethynyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-(Difluoromethyl)-9'-(neopentyloxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-(Difluoromethyl)-9'-(1 -methyl-1 H-pyrazol-4-yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-(Difluoromethyl)-2'-oxo-9'-(pyridin-3-yl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10-(Difluoromethyl)-6-isopropyl-6-methyl-9-(oxetan-3-ylmethoxy)-2-oxo-6,7-dihydro-2H- pyrido[2,1 -a]isoquinoline-3-carboxylic acid;
10-Cyclopropyl-6-isopropyl-9-methoxy-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 - a]isoquinoline-3-carboxylic acid;
10-(Difluoromethyl)-6-isopropyl-6-methyl-9-(1 -methyl-1 H-pyrazol-4-yl)-2-oxo-6,7-dihydro- 2H-pyrido[2,1 -a]isoquinoline-3-carboxylic acid;
10-(Difluoromethyl)-6-isopropyl-6-methyl-2-oxo-9-propyl-6,7-dihydro-2H-pyrido[2,1 - a]isoquinoline-3-carboxylic acid;
9'-(Oxetan-3-ylmethoxy)-2'-oxo-10'-(trifluoromethyl)-2',7'dihydrospiro
[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-chloro-9'-(3-methoxypropoxy)-3,3-dimethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-chloro-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclopentane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-chloro-9'-(cyclopropylmethoxy)-1 '-fluoro-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-chloro-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(3-hydroxypropoxy)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-chloro-9'-(cyclopropylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9- (Cyclopropylmethoxy)-10-(difluoromethyl)-1 -fluoro-6-isopropyl-6-methyl-2-oxo-6,7- dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylic acid;
9'-(Cyclopropylmethoxy)-10'-(difluoromethyl)-1 '-fluoro-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10- Methoxy-9-(3-methoxypropoxy)-6,6-dimethyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 - a]isoquinoline-3-carboxylic acid; 10'-Methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclopropane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(Benzyloxy)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclopentane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
10'-Methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclopentane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid;
9'-(Benzyloxy)-10'-methoxy-3,3-dimethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-Methoxy-9'-(3-methoxypropoxy)-3,3-dimethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10'-Borono-9'-(3-methoxypropoxy)-3,3-dimethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10"-Methoxy-9''-(3-methoxypropoxy)-2''-oxo-2'',7"-dihydrodispiro[cyclobutane-1 ,1 '- cyclobutane-3',6"-pyrido[2,1 -a]isoquinoline]-3"-carboxylic acid;
1 10'-Dichloro-9'-(3-methoxypropoxy)-3,3-dimethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid;
10-Cyclopropyl-9-(cyclopropylmethoxy)-6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H- pyrido[2,1 -a]isoquinoline-3-carboxylic acid;
6-lsopropyl-10-methoxy-9-(3-methoxypropoxy)-7 ,7-dimethyl-2-oxo-6,7-dihydro-2H- pyrido[2,1 -a]isoquinoline-3-carboxylic acid; and
9',10'-dimethoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylic acid; or a pharmaceutically acceptable salt thereof.
[0081] A particular embodiment provides a compound from Table 1 B, or a pharmaceutically acceptable salt thereof.
[0082] In particular embodiments, the present invention is directed to treatment the treatment of a hepatitis B infection or hepatitis B/hepatitis D co-infection in a human subject. Therefore, in certain embodiments, the present invention provides a method for the treatment of a hepatitis B infection or hepatitis B/hepatitis D co-infection, the method comprising administering to a subject in need of such treatment a compound of Formula I, IA or IB or a compound selected from Table 1A and Table 1 B as described herein.
[0083] In yet another embodiment, the present invention provides a method for the treatment of a hepatitis B infection or hepatitis B/hepatitis D co-infection, the method comprising administering to a subject in need of such treatment a first pharmaceutically acceptable agent comprising a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1A and Table 1 B as described as described herein, in combination with a second pharmaceutically acceptable agent that stimulates immune function and a third pharmaceutically acceptable agent comprising an antiviral compound.
[0084] In still other embodiments, the administration of a compound of Formula I, IA or IB or a compound selected from Table 1 A and Table 1 B as described as described herein inhibits the release of hepatitis B surface antigen (HBsAg), HB core antigen protein (HBcAg), and/or hepatitis B pre-core protein known as the HBV e-antigen antigen (HBeAg) from infected hepatocytes.
BRIEF DESCRIPTIONS OF THE DRAWINGS
[0085] Embodiments of the present invention features compounds that inhibit levels of HBe and/or HBs antigens in a subject infected with hepatitis B virus, and therefore are useful for treating human hepatitis B virus infections, and disease and symptoms associated with such virus infections. The foregoing features of the invention will be more readily understood by reference to the following detailed description, taken with reference to the accompanying Tables, in which:
[0086] Table 1 A and Table 1 B show a listing of compounds and structures, with ELISA data showing EC5o value ranges measured against the HBs antigen in a Hep AD 38 cell line (uM), for compounds from Table 1 A described herein.
DETAILED DESCRIPTION OF SPECIFIC EMBODIMENTS
[0087] Throughout this application, references are made to various embodiments relating to compounds, compositions, and methods. The various embodiments described are meant to provide a variety of illustrative examples and should not be construed as descriptions of alternative species. Rather it should be noted that the descriptions of various embodiments provided herein may be of overlapping scope. The embodiments discussed herein are merely illustrative and are not meant to limit the scope of the present invention.
[0088] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention. In this specification and in the claims that follow, reference will be made to a number of terms that shall be defined to have the following meanings.
[0089] As used herein unless otherwise specified, "alkyl" refers to a monovalent saturated aliphatic hydrocarbyl group having from 1 to 14 carbon atoms and, in some embodiments, from 1 to 6 carbon atoms The term "alkyl" includes, by way of example, linear and branched hydrocarbyl groups such as methyl (CH3-), ethyl (CH3CH2-), n-propyl
(CH3CH2CH2-), isopropyl ((CH3)2CH-), n-butyl (CH3CH2CH2CH2-), isobutyl ((CH3)2CHCH2-), sec-butyl ((CH3)(CH3CH2)CH-), /-butyl ((CH3)3C-), π-pentyl (CH3CH2CH2CH2CH2-), and neopentyl ((CH3)3CCH2-). AlkyI groups may also be substituted, for example, with one or more alkyl, cycloalkyl, heterocycloalkyl, alkoxy, carboxy, amido, keto, amino, cyano, aryl, heteroaryl, halo or haloalkyl substituents.
[0090] "Alkoxy" refers to the group -O-alkyl wherein alkyl is defined herein. Alkoxy includes, by way of example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, f-butoxy, sec-butoxy, n-pentoxy, morpholinylpropoxy, piperidinylethoxy.
[0091] "Amino" refers to the group -NRaRb where Ra and Rb are independently selected from hydrogen, alkyl, alkenyl, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, heterocyclic, and wherein Ra and Rb are optionally joined together with the nitrogen bound thereto to form a heterocyclic group. When Ra is hydrogen and Rb is alkyl, the amino group is sometimes referred to herein as alkylamino. When Ra and Rb are alkyl, the amino group is sometimes referred to herein as dialkylamino. When referring to a monosubstituted amino, it is meant that either Ra or Rb is hydrogen but not both. When referring to a disubstituted amino, it is meant that neither Ra nor Rb are hydrogen.
[0092] "Aryl" refers to an aromatic group of from 5 to 14 carbon atoms and no ring heteroatoms and having a single ring (e.g. , phenyl) or multiple condensed (fused) rings (e.g. , naphthyl or anthryl). For multiple ring systems, including fused, bridged, and spiro ring systems having aromatic and non-aromatic rings that have no ring heteroatoms, the term "Aryl" or "Ar" applies when the point of attachment is at an aromatic carbon atom (e.g. , 5,6,7,8 tetrahydronaphthalene-2-yl is an aryl group as its point of attachment is at the 2- position of the aromatic phenyl ring).
[0093] "Cycloalkyl" refers to a saturated or partially saturated cyclic group of from 3 to 14 carbon atoms and no ring heteroatoms and having a single ring or multiple rings including fused, bridged, and spiro ring systems. For multiple ring systems having aromatic and non-aromatic rings that have no ring heteroatoms, the term "cycloalkyl" applies when the point of attachment is at a non-aromatic carbon atom (e.g. 5,6,7,8,- tetrahydronaphthalene-5-yl). The term "Cycloalkyl" includes cycloalkenyl groups, such as cyclohexenyl. Examples of cycloalkyl groups include, for instance, adamantyl, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclooctyl, cyclopentenyl, and cyclohexenyl. Examples of cycloalkyl groups that include multiple bicycloalkyl ring systems are bicyclohexyl, bicyclopentyl, bicyclooctyl, and the like. Cycloalkyl groups may also be substituted, for example, with one or more alkyl, cycloalkyl, heterocycloalkyl, alkoxy, carboxy, amido, keto, amino, cyano, aryl, heteroaryl, halo or haloalkyl substituents.
[0094] "Halo" or "halogen" refers to fluoro, chloro, bromo, and iodo. [0095] "Haloalkyl" refers to substitution of alkyl groups with 1 to 9 (e.g. when the alkyl group has 3 carbon atoms, such as a t-butyl group fully-substituted with halogen) or in some embodiments 1 to 3 halo groups (e.g. trifluoromethyl).
[0096] "Hydroxy" or "hydroxyl" refers to the group -OH.
[0097] "Heteroaryl" refers to an aromatic group of from 1 to 14 carbon atoms and 1 to 6 heteroatoms selected from oxygen, nitrogen, sulfur, phosphorus, silicon and boron, and includes single ring (e.g. imidazolyl) and multiple ring systems (e.g. benzimidazol-2-yl and benzimidazol-6-yl). For multiple ring systems, including fused, bridged, and spiro ring systems having aromatic and non-aromatic rings, the term "heteroaryl" applies if there is at least one ring heteroatom and the point of attachment is at an atom of an aromatic ring (e.g. 1 ,2,3,4-tetrahydroquinolin-6-yl and 5,6,7,8-tetrahydroquinolin-3-yl). In some embodiments, the nitrogen and/or the sulfur ring atom(s) of the heteroaryl group are optionally oxidized to provide for the N-oxide (N→0), sulfinyl, or sulfonyl moieties. More specifically the term heteroaryl includes, but is not limited to, pyridyl, furanyl, thienyl, thiazolyl, isothiazolyl, triazolyl, imidazolyl, imidazolinyl, isoxazolyl, pyrrolyl, pyrazolyl, pyridazinyl, pyrimidinyl, purinyl, phthalazyl, naphthyl, naphthylpryidyl, oxazolyl, quinolyl, benzofuranyl,
tetrahydrobenzofuranyl, isobenzofuranyl, benzothiazolyl, benzoisothiazolyl, benzotriazolyl, indolyl, isoindolyl, indolizinyl, dihydroindolyl, indazolyl, indolinyl, benzoxazolyl, quinolyl, isoquinolyl, quinolizyl, quianazolyl, quinoxalyl, tetrahydroquinolinyl, isoquinolyl,
quinazolinonyl, benzimidazolyl, benzisoxazolyl, benzothienyl, benzopyridazinyl, pteridinyl, carbazolyl, carbolinyl, phenanthridinyl, acridinyl, phenanthrolinyl, phenazinyl, phenoxazinyl, phenothiazinyl, and phthalimidyl.
[0098] "Heterocyclic" or "heterocycle" or "heterocycloalkyl" or "heterocyclyl" refers to a saturated or partially saturated cyclic group having from 1 to 14 carbon atoms and from 1 to 6 heteroatoms selected from nitrogen, sulfur, phosphorus or oxygen and includes single ring and multiple ring systems including fused, bridged, and spiro ring systems. For multiple ring systems having aromatic and/or non-aromatic rings, the terms "heterocyclic",
"heterocycle", "heterocycloalkyl", or "heterocyclyl" apply when there is at least one ring heteroatom and the point of attachment is at an atom of a non-aromatic ring (e.g. 1 ,2,3,4- tetrahydroquinoline-3-yl, 5,6,7,8-tetrahydroquinoline-6-yl, and decahydroquinolin-6-yl). In one embodiment, the nitrogen, phosphorus and/or sulfur atom(s) of the heterocyclic group are optionally oxidized to provide for the N-oxide, phosphinane oxide, sulfinyl, sulfonyl moieties. More specifically the heterocyclyl includes, but is not limited to, tetrahydropyranyl, piperidinyl, piperazinyl, 3-pyrrolidinyl, 2-pyrrolidon-1 -yl, morpholinyl, and pyrrolidinyl. A prefix indicating the number of carbon atoms (e.g., C3-Ci0) refers to the total number of carbon atoms in the portion of the heterocyclyl group exclusive of the number of heteroatoms. [0099] Examples of heterocycle and heteroaryl groups include, but are not limited to, azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, pyridone, indolizine, isoindole, indole, dihydroindole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, naphthalene, oxazole, oxopyrrolidine, piperidine, piperazine, indoline, phthalimide, quinoline,
1 ,2,3,4-tetrahydroisoquinoline, 4,5,6,7-tetrahydrobenzo[b]thiophene, thiazole,
cyclopentathiazole, thiazolidine, thiophene, benzo[b]thiophene, morpholine, thiomorpholine (also referred to as thiamorpholine), piperidine, pyrrolidine, and tetrahydrofuranyl.
[00100] "Fused heterocyclic" refers to a 3 to 10 member cyclic substituent formed by the replacement of two hydrogen atoms at different carbon atoms in a cycloalkyl ring structure, as exemplified by the following cyclopentathiazole structure:
Figure imgf000029_0001
[00101] "Fused aryl and fused heteroaryl" refers to a 5 to 6 member aryl structure or heteroaryl structure fused with a 5- to 6- member aryl, heteroaryl or cycloalkyl ring at different carbon atoms in the aryl structure or the heteroaryl structure, which may be substituted at one of the carbons in the fused aryl or fused heteroaryl and connected to the core molecule at another of the carbons, as exemplified by the following cyclopentylthiazole, quinoline or naphthalene structures:
Figure imgf000029_0002
[00102] "Compound", "compounds", "chemical entity", and "chemical entities" as used herein refers to a compound encompassed by the generic formulae disclosed herein, any subgenus of those generic formulae, and any forms of the compounds within the generic and subgeneric formulae, including the racemates, stereoisomers, and tautomers of the compound or compounds.
[00103] The term "heteroatom" means nitrogen, oxygen, or sulfur and includes any oxidized form of nitrogen, such as N(O) {N +— O " } and sulfur such as S(O) and S(O) 2 , and the quaternized form of any basic nitrogen. [00104] "Oxazole" and "oxazolyl" refers to a 5-membered heterocyclic ring containing one nitrogen and one oxygen as heteroatoms and also contains three carbons and may be substituted at one of the three carbons and may be connected to another molecule at another of the three carbons, as exemplified by any of the following structures, wherein the oxazolidinone groups shown here are bonded to a parent molecule, which is indicated by a wavy line in the bond to the parent molecule:
Figure imgf000030_0001
[00105] "Oxopyrrolidine" and "oxopyrrolidinyl" refers to a 5-membered heterocyclic ring containing nitrogen and 4 carbons that is substituted at one of the carbons in the heterocyclic ring by a carbonyl and may be connected to another substituent at another carbon in the heterocyclic ring, as exem by the structure below:
Figure imgf000030_0002
[00106] "Pyridine" and "pyridinyl" refers to a 6-membered heteroaryl ring containing one nitrogen and 5 carbons that may also be substituted at one or more of the carbons in the heteroaryl ring, and may be connected to another substituent at another carbon in the heteroaryl ring, as exemplified by the structures below:
Figure imgf000030_0003
[00107] "Thiazole" and "thiazolyl" refers to a 5-membered heteroaryl containing one sulfur and one nitrogen in the heteroaryl ring and 3 carbons in the heteroaryl ring that may also be substituted at one or more of the carbons in the heteroaryl ring, and may be connected to another substituent at another carbon in the heteroaryl ring, as exemplified by the structures belo
Figure imgf000030_0004
[00108] "Pyrimidine" and "pyrimidinyl" refers to a 6-membered heteroaryl ring containing two nitrogens in the heteroaryl ring and 4 carbons in the heteroaryl ring that may be substituted at one or more of the carbons in the heteroaryl ring, and may be connected to another substituent at another carbon in the heteroaryl ring, as exemplified by the structures below:
Figure imgf000031_0001
[00109] "Racemates" refers to a mixture of enantiomers. In an embodiment of the invention, the compounds of Formulas I, or pharmaceutically acceptable salts thereof, are enantiomerically enriched with one enantiomer wherein all of the chiral carbons referred to are in one configuration. In general, reference to an enantiomerically enriched compound or salt, is meant to indicate that the specified enantiomer will comprise more than 50% by weight of the total weight of all enantiomers of the compound or salt.
[00110] "Solvate" or "solvates" of a compound refer to those compounds, as defined above, which are bound to a stoichiometric or non-stoichiometric amount of a solvent. Solvates of a compound includes solvates of all forms of the compound. In certain embodiments, solvents are volatile, non-toxic, and/or acceptable for administration to humans in trace amounts. Suitable solvates include water.
[00111] "Stereoisomer" or "stereoisomers" refer to compounds that differ in the chirality of one or more stereocenters. Stereoisomers include enantiomers and
diastereomers.
[00112] "Tautomer" refer to alternate forms of a compound that differ in the position of a proton, such as enol-keto and imine-enamine tautomers, or the tautomeric forms of heteroaryl groups containing a ring atom attached to both a ring -NH- moiety and a ring =N- moiety such as pyrazoles, imidazoles, benzimidazoles, triazoles, and tetrazoles.
[00113] "Pharmaceutically acceptable salt" refers to pharmaceutically acceptable salts derived from a variety of organic and inorganic counter ions well known in the art and include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, and tetraalkylammonium, and when the molecule contains a basic functionality, salts of organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, and oxalate. Suitable salts include those described in P. Heinrich Stahl, Camille G. Wermuth (Eds.), Handbook of Pharmaceutical Salts Properties, Selection, and Use; 2002.
[00114] "Patient" or "subject" refers to mammals and includes humans and non-human mammals. [00115] "Treating" or "treatment" of a disease in a patient refers to 1) preventing the disease from occurring in a patient that is predisposed or does not yet display symptoms of the disease; 2) inhibiting the disease or arresting its development; or 3) ameliorating or causing regression of the disease.
[00116] Wherever dashed lines occur adjacent to single bonds denoted by solid lines, then the dashed line represents an optional double bond at that position. Likewise, wherever dashed circles appear within ring structures denoted by solid lines or solid circles, then the dashed circles represent one to three optional double bonds arranged according to their proper valence taking into account whether the ring has any optional substitutions around the ring as will be known by one of skill in the art. For example, the dashed line in the structure below could either indicate a double bond at that position or a single bond at that position:
Figure imgf000032_0001
[00117] Similarly, ring A below could be a cyclohexyl ring without any double bonds or it could also be a phenyl ring having three double bonds arranged in any position that still depicts the proper valence for a phenyl ring. Likewise, in ring B below, any of X1-X5 could be selected from: C, CH, or CH2, N, or NH, and the dashed circle means that ring B could be a cyclohexyl or phenyl ring or a N-containing heterocycle with no double bonds or a N- containing heteroaryl ring with one to three double bonds arranged in any position that still depicts the proper valen
Figure imgf000032_0002
[00118] Where specific compounds or generic formulas are drawn that have aromatic rings, such as aryl or heteroaryl rings, then it will understood by one of still in the art that the particular aromatic location of any double bonds are a blend of equivalent positions even if they are drawn in different locations from compound to compound or from formula to formula. For example, in the two pyridine rings (A and B) below, the double bonds are drawn in different locations, however, they are known to be the same structure and compound:
Figure imgf000033_0001
[00119] Unless indicated otherwise, the nomenclature of substituents that are not explicitly defined herein are arrived at by naming the terminal portion of the functionality followed by the adjacent functionality toward the point of attachment. For example, the substituent "arylalkyloxycarbonyl" refers to the group (aryl)-(alkyl)-0-C(0)-. In a term such as "-CCR' , it should be understood that the two Rx groups can be the same, or they can be different if Rx is defined as having more than one possible identity. In addition, certain substituents are drawn as -RxRy, where the "-" indicates a bond adjacent to the parent molecule and Ry being the terminal portion of the functionality. Similarly, it is understood that the above definitions are not intended to include impermissible substitution patterns (e.g., methyl substituted with 5 fluoro groups). Such impermissible substitution patterns are well known to the skilled artisan.
Substitution with isotopes such as deuterium, i.e. 2H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances.
[00120] In accordance with one embodiment of the present invention, there is provided
a compound of Formula IA or Formula IB:
Figure imgf000033_0002
Formula I Formula IA Formula IB
[00121] wherein C* is a carbon atom stereocenter which has a configuration which is (R) or (S);
[00122] R1, R2, R3 and R4 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR20 R21 COR19; COOR19; CONR20 R21 ; pyrrolidinyl; unsubstituted Ci-6alkyl or halo- or alkyl substituted Ci_6alkyl; Ci-6alkylCOOR19; -BOR19OR19'; unsubstituted Ci-6alkoxy or halo-or alkyl-substituted Ci-6alkyoxy; unsubstituted C3-7cycloalkyl or halo-or alkyl-substituted C3-7cycloalkyl; unsubstituted C3-7cycloalkenyl or halo- or alkyl-substituted C3-7cycloalkenyl; unsubstituted or halo- or alkyl-substituted monocyclic heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O, N or S; unsubstituted or halo- or alkyl-substituted heteroaryl ring wherein a heteroatom in the heteroaryl ring is O, N or S; pyrrolidinyl; furanyl; pyrrolyl; imidazolyl; unsubstituted or halo- or alkyl-substituted -CxH2x- phenyl; unsubstituted or halo- or alkyl-substituted -0-CxH2x-phenyl; unsubstituted or halo- or alkyl-substituted -(Ci_6alkyl)N- CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; and - OR12; or R2 and R3 together form a 3 to 8 member cycloalkyi ring; or R2 and R3 together form a 3 to 8 member heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O, N, NR22, S, SR22 or SR22R22'; or R2 and R3 together form a 3 to 8 member aryl ring; or R2 and R3 together form a 3 to 8 member heteroaryl ring, wherein a heteroatom in the heteroaryl ring is O, N, NR22, S, SR22 or SR22R22';
[00123] and
[00124] (A) R5 and R6 are independently selected from halogen; cyano;
unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci-6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3-8cycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl; unsubstituted or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; and heteroaryl; or R5 and R6 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R13, R13', R14 and/or R14', or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R13, R13', R14 and/or R14', wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; and
[00125] R7 and R8 are independently hydrogen; halogen; cyano; unsubstituted Ci_ 6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3.
ecycloalkyl; unsubstituted or halo- or alkyl-substituted-CxH2x-phenyl; unsubstituted or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R7 and R8 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18', or R7 and R8 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R17, R17', R18 and/or R18', wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; or
[00126] (B) R5 and R8 are independently selected from hydrogen, unsubstituted Ci-6alkyl, and halo- or alkyl-substituted Ci_6alkyl; and
[00127] R6 and R7 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R15, R15', R16 and/or R16'; or R6 and R7 together form a 3- to 8- membered heterocycloalkyi ring comprising one heteroatom or two or more heteroatoms, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R15, R15', R16 and/or R16', wherein the one heteroatom in the heterocycloalkyi ring is NR20 and the two or more heteroatoms are selected from N, NR22, O, S, SR22 and SR22R22'; or
[00128] (C) R5 and R6 are independently selected from hydrogen; halogen; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci-6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl- substituted C3-8cycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl;
unsubstituted or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R5 and R6 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18', or R5 and R6 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R13 , R13', R14 and/or R14', wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; and
[00129] R7 and R8 are independently selected from halogen; cyano; unsubstituted Ci-6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3.
ecycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl; unsubstituted or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; and heteroaryl; or R7 and R8 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18', or R7 and R8 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R17 , R18', R17 and/or R18', wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; and
[00130] wherein
[00131] R9 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3.8cycloalkyl or halo- or alkyl-substituted C3.
ecycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted or halo- or alkyl-substituted-0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[00132] R10 is selected from COOR19and CONR20 R21 ;
[00133] R11 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_ 6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkyoxy; unsubstituted or halo- or alkyl-substituted C3.7cycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted or halo- or alkyl-substituted -0-CxH2)^phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[00134] R12 is selected from hydrogen; unsubstituted Ci_6alkyl or Ci-6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, heterocycloalkyi, heteroaryl, C3-8cycloalkyl and C2-6alkenyl; Ci-6alkylCi-6alkoxy; Ci-6alkylCi- 6alkoxyCi-6alkoxy; Ci.8alkylamino; Ci-8alkylaminocarbonylCi-6alkyl; Ci-6alkylaminosulfonylCi- 6alkyl; Ci_6alkylsulfonylCi-6alkyl; Ci-6alkylcyano; Ci-6alkylC3-8cycloalkyl; Ci-6alkylC3- ecycloalkylcyano; Ci-6alkylphenyl; Ci-6alkylcarbonylpyrrolidinyl; C2-6alkynyl; C2.6alkynylCi- 6hydroxy; Ci-6alkylCi-6alkoxyamino; Ci-6alkylCi-6alkoxyaminoCi-6alkyl; Ci_6alkylCi- 6alkoxyaminodiCi-6alkyl; Ci-6alkylcarboxy; Ci-6alkylaminocarbonylCi-6alkoxy; Ci_
6alkylheteroaryl, wherein the heteroaryl is an O-containing or N-containing monocyclic heteroaryl; or Ci-6alkylheterocycloalkyl, wherein the heterocycloalkyi is a monocyclic heterocycloalkyi;
[00135] R13 , R13', R14 and R14' are independently selected from hydrogen; halogen; amino; aminoalkyl; cyano; Ci_6alkyl; Ci-6alkoxy; carbonyl; carboxamide; and amide; or R13 and R13' or R14 and R14' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22';
[00136] R15, R15', R16 and R16' are independently hydrogen, halogen, amino, cyano, Ci-6alkyl, or Ci.6alkoxy, wherein any or R15, R15', R16 or R16', together with R5 or R8, form a 3- to 8-membered cycloalkyl ring or a 3-8 membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2.
6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22'; or any of R15, R15', R16 and/or R16' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3.8cycloalkyl, C2.6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
[00137] R17 , R17', R18 and R18' are independently selected from hydrogen; halogen; amino; cyano; Ci_6alkyl; and Ci-6alkoxy; or R17 and R18 or R17' and R18' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3.8cycloalkyl, C2.
6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22';
[00138] R19, R19' and R19" are independently selected from hydrogen; Ci_6alkyl; C3. ecycloalkyl; C2-6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci- 6alkyltetrazole; Ci-6alkylthiazole; Ci-6alkyloxazole; Ci-6alkyldioxazole; and Ci_
6alkyloxazolidone; and
[00139] R20 and R21 are independently selected from hydrogen; Ci_6alkyl; C3. ecycloalkyl; C2-6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci_
6alkyltetrazole; Ci-6alkylthiazole; Ci-6alkyloxazole; Ci-6alkyldioxazole; and Ci_
6alkyloxazolidone; or R20 and R21 together with the nitrogen to which they are attached form unsubstituted pyrrolidinyl, unsubstituted piperidinyl, or unsubstituted morpholinyl; or form carboxyl-substituted pyrrolidinyl, carboxyl-substituted piperidinyl or carboxyl-substituted morpholinyl; and
[00140] R22 and R22' are independently selected from hydrogen; oxygen; Ci_6alkyl; Ci-6haloalkyl; C3-8cycloalkyl; Ci-6alkylC3-8cycloalkyl; C2-6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci-6alkyltetrazole; Ci-6alkylthiazole; Ci-6alkyloxazole; Ci_ ealkyldioxazole; Ci-6alkyloxazolidone; -COR19; -COOR19'; -CSOR19 "; and -CONR20R21 ; and
[00141] R23 is selected from hydrogen; Ci_6alkyl; C3-8cycloalkyl; C2-6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci-6alkyltetrazole; Ci-6alkylthiazole; Ci_ 6alkyloxazole; Ci-6alkyldioxazole; Ci-6alkyloxazolidone; aryl; heteroaryl; and benzyl, or a pharmaceutically acceptable salt thereof.
[00142] One particular embodiment provides a compound according of Formula I, Formula IA or Formula IB as described herein, wherein:
[00143] R1 is selected from hydrogen, hydroxy, halogen, cyano, amino, pyrrolidinyl, unsubstituted Ci_6alkyl or halo-substituted Ci_6alkyl, unsubstituted Ci-6alkoxy or halo- substituted Ci-6alkyoxy; C3.7cycloalkyl or halo-substituted C3.7cycloalkyl; N-containing monocyclic heterocycloalkyi, pyrrolidinyl, -CxH2x-phenyl, -0-CxH2x-phenyl, or -(Ci_ 6alkyl)N- CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; or -OR12;
[00144] R2 and R3 are independently OR12;
[00145] R4 is selected from hydrogen, hydroxy, halogen, cyano, amino, pyrrolidinyl, unsubstituted Ci_6alkyl or halo-substituted Ci_6alkyl, unsubstituted Ci-6alkoxy or halo- substituted Ci-6alkyoxy; C3.7cycloalkyl or halo-substituted C3.7cycloalkyl; N-containing monocyclic heterocycloalkyi, pyrrolidinyl, -CxH2x-phenyl, -0-CxH2x-phenyl, or -(Ci_ 6alkyl)N- CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; or -OR12; and
R6 and R7 together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring comprising one heteroatom or two or more heteroatoms, optionally substituted with R15, R15', R16 and/or R16', wherein the one heteroatom in the heteroalkyl ring is NR20 and the two or more heteroatoms are selected from N, NR22, O, S, SR22 and SR22R22'; R11 is hydrogen; R12 is as described herein; and R15 and R16 or R15' and R16' together form a 3- to 8- membered cycloalkyi ring or heterocycloalkyi ring optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein the heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22'; or R15, R15', R16 or R16', together with R5 or R8, form a 3- to 8-membered cycloalkyi ring or a 3-8 membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
[00146] or a pharmaceutically acceptable salt thereof.
[00147] Another embodiment provides a compound of Formula I, Formula IA or Formula IB as described herein, wherein R10 is COOR19 or CONR20R21; or a
pharmaceutically acceptable salt thereof.
[00148] Another particular embodiment provides a compound of Formula I, Formula IA or Formula IB as described herein, wherein R10 is COOR19; R12 is unsubstituted Ci_6alkyl; and R19 is hydrogen, or a pharmaceutically acceptable salt thereof.
[00149] Another particular embodiment provides a compound of Formula I, Formula IA or Formula IB as described herein wherein R4 is hydrogen; R10 is COOR19; R12 is -CH3; and R19 is hydrogen, or a pharmaceutically acceptable salt thereof.
[00150] Another particular embodiment provides a compound of Formula I, Formula IA or Formula IB as described herein, wherein R2 and R3 are independently OR12; R5 and R6 together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with R13 and R14, wherein the heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22'; R10 is COOR19; R11 is hydrogen; R12 is unsubstituted Ci- 6alkyl; R13 and R14 or R13' and R14' together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein the heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22'; and R19 is hydrogen, or a pharmaceutically acceptable salt thereof.
[00151] Still another particular embodiment provides a compound of Formula I, Formula IA or Formula IB as described herein, wherein R2 and R3 are independently OR12; R7 and R8 together form a 3- to 8-membered ring, optionally substituted with R17 and R18; R10 is COOR19; R11 is hydrogen; R12 is unsubstituted Ci_6alkyl; R17 and R18 or R17' and R18' together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3. ecycloalkyl, C2-6alkenyl or Ci.6alkoxy, wherein the heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22'; and R19 is hydrogen, or a pharmaceutically acceptable salt thereof.
[00152] Another particular embodiment provides a compound of Formula I, Formula IA or Formula IB as described herein, wherein R2 and R3 are independently OR12; and R5 and R6 together form a 3- to 8- membered cycloalkyl ring or 3- to 8- membered
heterocycloalkyl ring, optionally substituted with R13 , R14, R13' and/or R14', wherein the heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22', or a pharmaceutically acceptable salt thereof.
[00153] Another particular embodiment provides a compound of Formula I, Formula IA or Formula IB as described herein wherein R2 and R3 are independently OR12; and R7 and R8 together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered
heterocycloalkyl ring, optionally substituted with R17, R18, R17' and/or R18', wherein the heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22', or a pharmaceutically acceptable salt thereof.
[00154] In accordance with one embodiment of the present invention, there is provided a compound having the structure of Formula I, IA or IB wherein the compounds are selected from:
Figure imgf000039_0001
R
O O , aminosulfonamide O O t heterocycle, heteroaryl.
[00156] In particular embodiments there are provided compounds of Formula I, IA or IB wherein the compounds are selected from:
Figure imgf000039_0002
[00157] In other particular embodiments there are provided compounds of Formula I, IA or IB wherein the compounds are selected from:
Figure imgf000040_0001
wherein R is H or alkyl.
[00158] In another embodiment of the present invention, compounds of Formula I, Formula IA or Formula IB are selected from:
Figure imgf000040_0002
Figure imgf000040_0003
wherein R is hydrogen or alkyi and R' is H, alkyl, ester, carbamate, urea, or sulfonyl urea. [00159] In another embodiment of the present invention there is provided a compound of Formula I, Formula IA or Formula IB as shown:
Figure imgf000041_0001
[00160] wherein R1 , R2, R3, R4, R9, R10 and R11 are as described, and R' and R are independently selected from hydrogen, halo, alkyl, aryl, carbocycle, heterocycle, heteroaryl, alkylene optionally substituted with
',
Figure imgf000041_0002
combine to form a spiro, fused or bridged carbocylic or heterocyclic ring.
[00162] In another embodiment of the present invention, there is provided a compound having the structure of Formula I, IA or IB, wherein the compound is selected
Figure imgf000041_0003
wherein R and R' are independently H, alkyl, acyl, ester, carbamoyl, sulfonyl urea, or urea, and wherein Z is substituted C, O, N or S.
[00163] In another particular embodiment, there is provided a compound of Formula I, IA or IB as indicated:
Figure imgf000042_0001
wherein R4, R5, R6, R9, R10 and R11 are as described, and R1 is halogen, R2 and R3 are independently selected from:
R2 R3
Figure imgf000043_0001
Figure imgf000043_0002
Ck
Δ,
CI--..
F...._
Me'
BO"
Ek
Pr-.
[00164] In a particular embodiment of the invention, there is provided a compound selected from Table 1 A and Table 1 B , or a pharmaceutically acceptable salt thereof.
[00165] Compounds described herein can exist in particular geometric or stereoisomeric forms. The invention contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)-isomers, the racemic mixtures thereof, and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, as falling within the scope of the invention. Additional asymmetric carbon atoms can be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in this invention.
[00166] Optically active (R)- and (S)-isomers and d and I isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. If, for instance, a particular enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis, or by derivatization with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers. Alternatively, where the molecule contains a basic functional group, such as an amino group, or an acidic functional group, such as a carboxyl group, diastereomeric salts can be formed with an appropriate optically active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographic means known in the art, and subsequent recovery of the pure
enantiomers. In addition, separation of enantiomers and diastereomers is frequently accomplished using chromatography employing chiral, stationary phases, optionally in combination with chemical derivatization (e.g., formation of carbamates from amines).
[00167] In another embodiment of the invention, there is provided a compound of Formula I, Formula IA or Formula IB or a pharmaceutically acceptable salt thereof selected from Table 1 A and 1 B, for use in therapy.
[00168] In another embodiment of the invention, there is provided a compound of Formula I, Formula IA or Formula IB, or a compound selected from Table 1 A and Table 1 B, or a pharmaceutically acceptable salt thereof, for use in the treatment of a viral infection.
[00169] In another embodiment of the invention, there is provided a use of a compound of Formula I, Formula IA or Formula IB, or a compound selected from Table 1 A and Table 1 B, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for use in the treatment of a viral infection in a human.
[00170] In another embodiment of the invention, there is provided a pharmaceutical composition comprising a pharmaceutically acceptable diluent and a therapeutically effective amount of a compound of Formula I, Formula IA or Formula IB, or a compound selected from Table 1 A and Table 1 B, or a pharmaceutically acceptable salt thereof.
[00171] In one embodiment, the pharmaceutical formulation containing a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1 A and Table 1 B or a pharmaceutically acceptable salt thereof is a formulation adapted for parenteral administration. In another embodiment, the formulation is a long-acting parenteral formulation. In a further embodiment, the formulation is a nano-particle formulation.
[00172] In one embodiment, the pharmaceutical formulation containing a compound of Formula I, Formula IA or Formula IB, or a compound selected from Table 1 A and Table 1 B, or a pharmaceutically acceptable salt thereof is a formulation adapted for oral, rectal, topical or intravenous formulation, wherein the pharmaceutical formulation optionally comprises any one or more of a pharmaceutically acceptable carrier, adjuvant or vehicle.
[00173] In one embodiment, the compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1A and Table 1 B or a pharmaceutically acceptable salt thereof is formulated for oral administration, and can be administered as a conventional preparation, for example, as any dosage form of a solid agent such as tablets, powders, granules, capsules and the like; an aqueous agent; an oily suspension; or a liquid agent such as syrup and elixir. In one embodiment, the compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1 A and Table 1 B or a pharmaceutically acceptable salt thereof is formulated for parenteral administration, and can be administered as an aqueous or oily suspension injectable, or a nasal drop. Upon preparation of a parenteral formulation with a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1A and Table 1 B or a pharmaceutically acceptable salt thereof, conventional excipients, binders, lubricants, aqueous solvents, oily solvents, emulsifiers, suspending agents, preservatives, stabilizers and the like may be arbitrarily used. As an anti-viral-drug, particularly, an oral agent is preferable. A preparation of a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1A and Table 1 B or a pharmaceutically acceptable salt thereof may be prepared by combining (e.g. mixing) a therapeutically effective amount of a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1 A and Table 1 B or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable carrier or diluent.
[00174] Pharmaceutical formulations adapted for oral administration may be presented as discrete units such as capsules or tablets; powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; edible foams or whips; or oil-in-water liquid emulsions or water-in-oil liquid emulsions.
[00175] For instance, for oral administration in the form of a tablet or capsule, the compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1A and Table 1 B can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. Powders are prepared by comminuting the compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1 A and Table 1 B to a suitable fine size and mixing with a similarly comminuted pharmaceutical carrier such as an edible carbohydrate, as, for example, starch or mannitol. Flavoring, preservative, dispersing and coloring agent can also be present.
[00176] Capsules are made by preparing a powder mixture, as described above, and filling formed gelatin sheaths. Glidants and lubricants such as colloidal silica, talc, magnesium stearate, calcium stearate or solid polyethylene glycol can be added to the powder mixture before the filling operation. A disintegrating or solubilizing agent such as agar-agar, calcium carbonate or sodium carbonate can also be added to improve the availability of the medicament when the capsule is ingested. [00177] Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. Tablets are formulated, for example, by preparing a powder mixture, granulating or slugging, adding a lubricant and disintegrant and pressing into tablets. A powder mixture is prepared by mixing the compound, suitably comminuted, with a diluent or base as described above, and optionally, with a binder such as carboxymethylcellulose, an aliginate, gelatin, or polyvinyl pyrrolidone, a solution retardant such as paraffin, a resorption accelerator such as a quaternary salt and/or an absorption agent such as bentonite, kaolin or dicalcium phosphate. The powder mixture can be granulated by wetting with a binder such as syrup, starch paste, acadia mucilage or solutions of cellulosic or polymeric materials and forcing through a screen. As an alternative to granulating, the powder mixture can be run through the tablet machine and the result is imperfectly formed slugs broken into granules. The granules can be lubricated to prevent sticking to the tablet forming dies by means of the addition of stearic acid, a stearate salt, talc or mineral oil. The lubricated mixture is then compressed into tablets. The compounds of the present invention can also be combined with a free flowing inert carrier and compressed into tablets directly without going through the granulating or slugging steps. A clear or opaque protective coating consisting of a sealing coat of shellac, a coating of sugar or polymeric material and a polish coating of wax can be provided. Dyestuffs can be added to these coatings to distinguish different unit dosages.
[00178] Oral fluids such as solutions, syrups and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of the compound. Syrups can be prepared by dissolving the compound in a suitably flavored aqueous solution, while elixirs are prepared through the use of a non-toxic alcoholic vehicle. Suspensions can be formulated by dispersing the compound in a non-toxic vehicle. Solubilizers and emulsifiers such as ethoxylated isostearyl alcohols and polyoxy ethylene sorbitol ethers, preservatives, flavor additive such as peppermint oil or natural sweeteners or saccharin or other artificial sweeteners, and the like can also be added.
[00179] Where appropriate, dosage unit formulations for oral administration can be microencapsulated. The formulations of compounds of Formula I, Formula IA or Formula IB can also be prepared to prolong or sustain the release of the compound, as for example by coating or embedding particulate material in polymers, wax or the like.
[00180] The compounds of Formula I, Formula IA or Formula IB or compounds selected from Table 1A and Table 1 B, or salts, solvates or hydrates thereof, can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.
[00181] The compounds of Formula I, Formula IA or Formula IB or compounds selected from Table 1 A and Table 1 B, or salts, solvates or hydrates thereof, may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled. The compounds may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyrancopolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxyethylaspartamide-phenol, or polyethyleneoxidepolylysine substituted with palmitoyl residues. Furthermore, the compounds may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked or amphipathic block copolymers of hydrogels.
[00182] Pharmaceutical formulations adapted for transdermal administration may be presented as discrete patches intended to remain in intimate contact with the epidermis of the recipient for a prolonged period of time. For example, the compounds of Formula I, Formula IA or Formula IB or compounds selected from Table 1 A and Table 1 B may be delivered from a patch by iontophoresis as generally described in Pharmaceutical Research, 3(6), 318 (1986).
[00183] Pharmaceutical formulations adapted for topical administration may be formulated as ointments, creams, suspensions, lotions, powders, solutions, pastes, gels, sprays, aerosols or oils. When formulated in an ointment, the active ingredient may be employed with either a paraffinic or a water-miscible ointment base. Alternatively, the active ingredient may be formulated in a cream with an oil-in-water cream base or a water-in-oil base.
[00184] Pharmaceutical formulations adapted for rectal administration may be presented as suppositories or as enemas.
[00185] Pharmaceutical formulations adapted for nasal administration wherein the carrier is a solid include a coarse powder having a particle size for example in the range 20 to 500 microns which is administered in the manner in which snuff is taken, i.e. by rapid inhalation through the nasal passage from a container of the powder held close up to the nose. Suitable formulations wherein the carrier is a liquid, for administration as a nasal spray or as nasal drops, include aqueous or oil solutions of the active ingredient.
[00186] Pharmaceutical formulations adapted for administration by inhalation include fine particle dusts or mists, which may be generated by means of various types of metered, dose pressurized aerosols, nebulizers or insufflators.
[00187] Pharmaceutical formulations adapted for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The formulations may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets.
[00188] It should be understood that in addition to the ingredients particularly mentioned above, the formulations described herein may include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavouring agents.
[00189] A therapeutically effective amount of a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1A and Table 1 B will depend upon a number of factors including, for example, the age and weight of the human or other animal, the precise condition requiring treatment and its severity, the nature of the formulation, and the route of administration, and will ultimately be at the discretion of the attendant physician or veterinarian. An effective amount of a salt or hydrate thereof may be determined as a proportion of the effective amount of the compound of Formula I, Formula IA or Formula IB or the compound selected from Table 1 A and Table 1 B, or salts, solvates or hydrates thereof, per se.
[00190] Embodiments of the present invention provide administration of a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1 A and Table 1 B to a healthy or virus-infected patient, either as a single agent or in combination with (a) another agent that is effective in treating or preventing hepatitis B virus of hepatitis D virus, (b) another agent that improves immune response and robustness, or (c) another agent that reduces inflammation and/or pain.
[00191] The compounds of Formula I, Formula IA or Formula IB or compounds selected from Table 1 A and Table 1 B, or salts, solvates or hydrates thereof, are believed to have activity in preventing, halting or reducing the effects of hepatitis B virus by inhibiting the HBe and/or HBs antigens, thereby interfering with or preventing the virus from remaining in the host cell and rendering the virus unable to replicate.
[00192] Accordingly, there is provided a method of treating a hepatitis B virus and/or hepatitis D virus by administering a therapeutically effective amount of a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1 A and Table 1 B, or a pharmaceutically acceptable salt, solvate or hydrate thereof, to a mammal. In one embodiment, the mammal is a human.
[00193] In another aspect of the present invention, there is provided a method of reducing HBe and/or HBs antigens in a mammal by administering to said mammal a therapeutically effective amount of a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1A and Table 1 B or a pharmaceutically acceptable salt, solvate or hydrate thereof. In one embodiment, the mammal is a human.
[00194] In other embodiments, the compounds of the present invention may be used in combination with one or more antiviral therapeutic agents or anti-inflammatory agents useful in the prevention or treatment of viral diseases or associated pathophysiology. Thus, the compounds of the present invention and their salts, solvates, or other pharmaceutically acceptable derivatives thereof, may be employed alone or in combination with other antiviral or anti-inflammatory therapeutic agents.
[00195] The compounds of the present invention and any other pharmaceutically active agent(s) may be administered together or separately and, when administered separately, administration may occur simultaneously or sequentially, in any order. The amounts of the compounds of the present invention and the other pharmaceutically active agent(s) and the relative timings of administration will be selected in order to achieve the desired combined therapeutic effect. The administration in combination of a compound of the present invention and salts, solvates, or other pharmaceutically acceptable derivatives thereof with other treatment agents may be in combination by administration concomitantly in: (1) a unitary pharmaceutical composition including both compounds; or (2) separate pharmaceutical compositions each including one of the compounds.
[00196] Alternatively, the combination may be administered separately in a sequential manner wherein one treatment agent is administered first and the other second or vice versa. Such sequential administration may be close in time or remote in time. The amounts of the compound(s) of Formula I, Formula IA or Formula IB or compounds selected from Table 1 A and Table 1 B or salts thereof, and the other pharmaceutically active agent(s), and the relative timings of administration, will be selected in order to achieve the desired combined therapeutic effect. [00197] More particularly, embodiments provide a method as described comprising administering an additional agent selected from an antiviral agent, an antibiotic, an analgesic, a non-steroidal anti-inflammatory (NSAID) agent, an antifungal agent, an antiparasitic agent, an anti-nausea agent, an anti-diarrheal agent, or an immunosuppressant agent. In certain embodiments, the antiviral agent is an anti-hepatitis B agent or an anti- hepatitis C agent. Still more particularly, the additional agent is administered as part of a single dosage form of said pharmaceutical formulation, or as a separate dosage form.
[00198] The present invention is directed to compounds, compositions and pharmaceutical compositions that have utility as novel treatments and/or preventative therapies for virus infections. While not wanting to be bound by any particular theory, it is thought that the present compounds are able to inhibit the levels of HBe and HBs antigens in a subject infected with hepatitis B virus or suffering from a chronic hepatitis B viral infection. By reducing the levels of HBe and HBs antigens in a subject infected with hepatitis B virus, compounds described herein are effective at treating hepatitis B infections, and secondary disorders such as liver cirrhosis, liver failure and liver cancer which are often associated with hepatitis B virus infections.
[00199] Therefore, in another embodiment of the present invention, there is provided a method of treating or preventing a hepatitis B virus infection in a subject suffering from the HBV infection comprising administering to the subject an inhibitor of HBe and/or HBs antigens, wherein the inhibitor is a compound of Formula I, Formula IA or Formula IB.
[00200] In another embodiment of the present invention, there is provided a method of treating a hepatitis B virus infection and/or a hepatitis D virus infection in a subject suffering from the virus infection comprising administering to the subject a compound from Table 1 A or a compound from Table 1 B .
[00201] In some embodiments, provided is a method for treating a viral infection in a subject mediated at least in part by a virus in the hepatitis B family and or the hepatitis D family, comprising administering to the subject a composition comprising a compound of any of Formula I, Formula IA or Formula IB, or a compound selected from Table 1 A and Table 1 B, or a pharmaceutically acceptable salt thereof.
[00202] In yet another aspect, another embodiment of the present invention provides a method of inhibiting progression of a viral infection in a subject at risk for infection with a hepatitis B virus and/or a hepatitis D virus, comprising administering to the subject a therapeutically effective amount of the compound of Formula I, Formula IA or Formula IB, or a compound selected from Table 1A and Table 1 B, or a pharmaceutically acceptable salt thereof. [00203] In yet another aspect, another embodiment of the present invention provides a method of preventing a viral infection in a subject at risk for infection from a hepatitis B virus and/or a hepatitis D virus comprising administering to the subject a therapeutically effective amount of the compound of Formula I or Formula IA, or a compound selected from Table 1 A and Table 1 B, or a pharmaceutically acceptable salt thereof.
[00204] In yet another aspect, another embodiment of the present invention provides a method of treating a virus infection in a subject suffering from said virus infection, comprising administering to the subject a therapeutically effective amount of the compound of Formula I, Formula IA or Formula IB, or a compound selected from Table 1 A and Table 1 B, or a pharmaceutically acceptable salt thereof.
[00205] Compounds, methods and pharmaceutical compositions for treating viral infections, by administering a compound of Formula I, Formula IA or Formula IB or a compound selected from Table 1A and Table 1 B in a therapeutically effective amount are disclosed. Methods for preparing compounds of Formula I, Formula IA or Formula IB or compounds selected from Table 1A and Table 1 B and methods of using the compounds and pharmaceutical compositions thereof are also disclosed. In particular, the treatment and prophylaxis of viral infections such as those caused by hepatitis B and/or hepatitis D are disclosed.
[00206] In other embodiments, the compounds described herein are useful for treating infections in a subject wherein the infection is caused by a multi-drug resistant strain of the hepatitis B virus and/or a hepatitis D virus.
[00207] In further embodiments, the compound of the present invention, or a pharmaceutically acceptable salt thereof, is chosen from the compounds set forth in Table 1A and Table 1 B.
EXAMPLES
[00208] Synthetic Schemes
[00209] The compounds of the present invention having Formula I, Formula IA or
Figure imgf000051_0001
Formula I Formula IA Formula IB, [00210] or corresponding pharmaceutically acceptable salts thereof, are prepared using conventional organic syntheses, wherein:
[00211] C* is a carbon atom stereocenter which has a configuration which is (R) or
(S);
[00212] R1, R2, R3 and R4 are independently selected from hydrogen, hydroxy, halogen, cyano, amino, pyrrolidinyl, unsubstituted Ci_6alkyl or halo-substituted Ci_6alkyl, unsubstituted Ci-6alkoxy or halo-substituted Ci-6alkyoxy; C3-7cycloalkyl or halo-substituted C3-7cycloalkyl; N-containing monocyclic heterocycloalkyi, pyrrolidinyl, -CxH2x-phenyl, -O- CxH2x-phenyl or -(Ci-6alkyl)N- CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; or -OR12; and wherein
[00213] (A) R5 and R6 are independently halogen, cyano, unsubstituted Ci_6alkyl or halo-substituted Ci_6alkyl, unsubstituted Ci-6alkoxy or halo-substituted Ci-6alkoxy, unsubstituted C3-8cycloalkyl or halo-substituted C3-8cycloalkyl, -CxH2x-phenyl or -0-CxH2x- phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; heteroaryl; or R5 and R6 together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with R13, R13', R14 and/or R14', wherein the heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; and
[00214] R7 and R8 are independently hydrogen, halogen, cyano, unsubstituted Ci_ 6alkyl or halo-substituted Ci_6alkyl, unsubstituted Ci-6alkoxy or halo-substituted Ci-6alkoxy, unsubstituted C3.8cycloalkyl or halo-substituted C3.8cycloalkyl, -CxH2x-phenyl or -0-CxH2x- phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; heteroaryl; or R7 and R8 together form a 3- to 8- membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with R17, R17', R18 and/or R18', wherein the heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; or
[00215] (B) R5 is selected from hydrogen, unsubstituted Ci_6alkyl, and halo- or alkyl-substituted Ci_6alkyl;
[00216] R6 and R7 together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring comprising one heteroatom or two or more heteroatoms, optionally substituted with R15, R15', R16 and/or R16', wherein the one heteroatom in the heteroalkyl ring is NR20 and the two or more heteroatoms are selected from N, NR22, O, S, SR22 and SR22R22'; and
[00217] R8 is selected from hydrogen, unsubstituted Ci_6alkyl, and halo- or alkyl- substituted Ci-6alkyl; or
[00218] (C) R5 and R6 are independently hydrogen, halogen, cyano,
unsubstituted Ci_6alkyl or halo-substituted Ci_6alkyl, unsubstituted Ci-6alkoxy or halo- substituted Ci-6alkoxy, unsubstituted C3.8cycloalkyl or halo-substituted C3.8cycloalkyl, - CxH2x-phenyl or -0-CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; heteroaryl; or R5 and R6 together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered
heterocycloalkyi ring, optionally substituted with R13 , R13', R14 and/or R14', wherein the heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; and
[00219] R7 and R8 are independently halogen, cyano, unsubstituted Ci_6alkyl or halo- substituted Ci-6alkyl, unsubstituted Ci-6alkoxy or halo-substituted Ci-6alkoxy, unsubstituted C3-8cycloalkyl or halo-substituted C3-8cycloalkyl, -CxH2x-phenyl or -0-CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; heteroaryl; or R7 and R8 together form a 3- to 8- membered cycloalkyi ring or a 3- to 8- membered heterocycloalkyi ring, optionally substituted with R17 , R18', R17 and/or R18', wherein the heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22';
and wherein
[00220] R9 is hydrogen, hydroxy, halogen, cyano, unsubstituted Ci_6alkyl or halo- substituted Ci-6alkyl, unsubstituted Ci-6alkoxy or halo-substituted Ci-6alkoxy, unsubstituted C3-8cycloalkyl or halo-substituted C3-8cycloalkyl, -CxH2x-phenyl or -0-CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[00221] R10 is hydrogen; hydroxy; halogen; cyano; COOR19, CONR20 R21, unsubstituted Ci_6alkyl or halo-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- substituted Ci-6alkyoxy; C3.7cycloalkyl; -CxH2x-phenyl or -0-CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[00222] R11 is hydrogen, hydroxy, halogen, cyano, unsubstituted Ci_6alkyl or halo- substituted Ci-6alkyl, unsubstituted Ci-6alkoxy or halo-substituted Ci-6alkyoxy; C3.7cycloalkyl; -CxH2x-phenyl or -0-CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6;
[00223] R12 is hydrogen; unsubstituted Ci_6alkyl or Ci_6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, C3.8cycloalkyl and C2. 6alkenyl; Ci-6alkylCi-6alkoxy; Ci-6alkylCi-6alkoxyCi-6alkoxy; Ci-8alkylamino; Ci_
8alkylaminocarbonylCi-6alkyl; Ci-6alkylaminosulfonylCi-6alkyl; Ci-6alkylsulfonylCi_6alkyl; Ci_ 6alkylcyano; Ci-6alkylC3-8cycloalkyl; Ci-6alkylC3-8cycloalkylcyano; Ci-6alkylphenyl; Ci- 6alkylcarbonylpyrrolidinyl; C2.6alkynyl; C2-6alkynylCi-6hydroxy; Ci-6alkylCi-6alkoxyamino; Ci_ 6alkylCi-6alkoxyaminoCi-6alkyl; Ci-6alkylCi-6alkoxyaminodiCi-6alkyl; Ci-6alkylcarboxy; or Ci_ 6alkylaminocarbonylCi-6alkoxy; Ci-6alkylheteroaryl, wherein heteroaryl is N-containing monocyclic heteroaryl; or Ci-6alkylheterocycloalkyl, wherein heterocycloalkyi is monocyclic heterocycloalkyi;
[00224] R13 , R13', R14 and R14' are independently hydrogen, halogen, amino, aminoalkyl, cyano, Ci_6alkyl, Ci-6alkoxy, carbonyl, carboxamide, amide; or R13 and R13' or R14 and R14' together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci-6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein the heteroatom in the
heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22';
[00225] R15, R15', R16 and R16' are independently hydrogen, halogen, amino, cyano, Ci-6alkyl, or Ci-6alkoxy, wherein any of R15, R15', R16 or R16', together with R5 or R8, form a 3- to 8-membered cycloalkyi ring or a 3-8 membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2.
6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22'; or any of R15, R15', R16 and/or R16' together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
[00226] R17 , R17', R18 and R18' are independently hydrogen, halogen, amino, cyano, Ci-6alkyl, or Ci-6alkoxy; or R17 and R18 or R17' and R18' together form a 3- to 8-membered cycloalkyi or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein the heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22';
[00227] R19, R19' and R19 " are independently hydrogen, Ci_6alkyl, C3-8cycloalkyl, C2. 6alkenyl, phenyl, Ci-6alkylimidizole, Ci-6alkoxy, Ci-6alkyltriazole, Ci-6alkyltetrazole, Ci_
6alkylthiazole, Ci-6alkyloxazole, Ci-6alkyldioxazole; Ci-6alkyloxazolidone; and
[00228] R20 and R21 are independently hydrogen, Ci_6alkyl, C3-8cycloalkyl, C2. 6alkenyl, phenyl, Ci-6alkylimidizole, Ci-6alkoxy, Ci-6alkyltriazole,Ci-6alkyltetrazole, Ci_
6alkylthiazole, Ci-6alkyloxazole, Ci-6alkyldioxazole; Ci-6alkyloxazolidone, or R20 and R21 together with the nitrogen to which they are attached form unsubstituted pyrrolidinyl, unsubstituted piperidinyl, or unsubstituted morpholinyl; or form carboxyl-substituted pyrrolidinyl, carboxyl-substituted piperidinyl or carboxyl-substituted morpholinyl; and
[00229] R22 and R22' are independently selected from hydrogen, oxygen, Ci_6alkyl, C3-ecycloalkyl, C2-6alkenyl, phenyl, Ci-6alkylimidizole, Ci_6alkoxy, Ci-6alkyltriazole,Ci- 6alkyltetrazole, Ci-6alkylthiazole, Ci-6alkyloxazole, Ci-6alkyldioxazole; Ci-6alkyloxazolidone, - COR19, -COOR19', -CSOR19 ", -CONR20R21 , or a pharmaceutically acceptable salt thereof.
[00230] One particular embodiment provides a compound according of Formula I, Formula IA or Formula IB as described herein, wherein:
[00231] R1 is selected from hydrogen, hydroxy, halogen, cyano, amino, pyrrolidinyl, unsubstituted Ci_6alkyl or halo-substituted Ci_6alkyl, unsubstituted Ci-6alkoxy or halo- substituted Ci-6alkyoxy; C3.7cycloalkyl or halo-substituted C3.7cycloalkyl; N-containing monocyclic heterocycloalkyl, pyrrolidinyl,
Figure imgf000055_0001
-0-CxH2x-phenyl, or -(Ci_ 6alkyl)N- CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; or -OR12;
[00232] R2 and R3 are independently OR12;
[00233] R4 is selected from hydrogen, hydroxy, halogen, cyano, amino, pyrrolidinyl, unsubstituted Ci_6alkyl or halo-substituted Ci_6alkyl, unsubstituted Ci-6alkoxy or halo- substituted Ci-6alkyoxy; C3-7cycloalkyl or halo-substituted C3-7cycloalkyl; N-containing monocyclic heterocycloalkyl, pyrrolidinyl, -CxH2x-phenyl, -0-CxH2x-phenyl, or -(Ci_ 6alkyl)N- CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; or -OR12;
[00234] R6 and R7 together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyl ring, optionally substituted with R15 , R15', R16 and/or R16', wherein the heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22'; R9 is hydrogen; R11 is hydrogen; R12 is as described herein; and R15 and R16 or R15' and R16' together form a 3- to 8-membered cycloalkyl ring or heterocycloalkyl ring optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2. 6alkenyl or Ci-6alkoxy, wherein the heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22', or a pharmaceutically acceptable salt thereof.
[00235] Suitable synthetic routes are depicted below in the following general reaction schemes. The skilled artisan will appreciate that if a substituent described herein is not compatible with the synthetic methods described herein, the substituent may be protected with a suitable protecting group that is stable to the reaction conditions. The protecting group may be removed at a suitable point in the reaction sequence to provide a desired intermediate or target compound. Suitable protecting groups and the methods for protecting and de-protecting different substituents using such suitable protecting groups are well known to those skilled in the art; examples of which may be found in T. Greene and P. Wuts, Protecting Groups in Chemical Synthesis (3rd ed.), John Wiley & Sons, NY (1999). In some instances, a substituent may be specifically selected to be reactive under the reaction conditions used. Under these circumstances, the reaction conditions convert the selected substituent into another substituent that is either useful as an intermediate compound or is a desired substituent in a target compound.
[00236] Abbreviations
[00237] In describing the examples, chemical elements are identified in accordance with the Periodic Table of the Elements. Abbreviations and symbols utilized herein are in [00238] accordance with the common usage of such abbreviations and symbols by those skilled in the chemical arts. The following abbreviations are used herein:
AcOH acetic acid
Ac20 acetic anhydride
aq aqueous
B4 (S)-2-(((benzyloxy)carbonyl)amino)-3-(4-(trifluoromethyl)piperidin-1 - yl)propanoic acid
BOC (Boc) N-tert-butoxycarbonyl or tert-butyloxycarbonyl
CBz carboxybenzyl
dba dibenzylideneacetone or dibenzalacetone
DCE dichloroethane
DCM dichloromethane
DCM/EA dichloromethane/ethanol
DDQ 2,3-dichloro-5,6-dicyanobenzoquinone
DIPEA (or DIEA) Ν,Ν-diisopropylethylamine, or Hiinig's base
DME dimethoxyethane
DMEM Dulbecco's Modified Eagle Medium
DMF dimethylformamide
DMP Dess-Martin periodinane
DMSO-d6 deuterated dimethylsulfoxide
DMSO dimethylsulfoxide
DPPA diphenyl phosphoryl azide
ECso 50% effective concentration
EDTA ethylenediaminetetraacetic acid
Et ethyl
Et20 diethyl ether
EtOH ethanol
EtOAc, EA, AcOEt ethyl acetate
GlutaMAX™ cell culture supplement from Life Technologies
h hour(s)
HEPES 4-(2-hydroxyethyl)-1 -piperazineethanesulfonic acid
HPLC high performance liquid chromatography
IC50 50% inhibition concentration
iPrOH isopropyl alcohol or isopropanol
LCMS Liquid chromatography mass spectroscopy
LDA lithium di-isopropyl amide
Me methyl
MeOH methanol
NBS N-bromosuccinimide
NCS N-chlorosuccinimide
NIS N-iodosuccinimide NXS N-halosuccinimide
NaBH(OAc)3 sodium triacetoxyborohydride
NMR Nuclear Magnetic Resonance spectroscopy
Pd2(dba)3 Tris(dibenzylideneacetone)dipalladium(0)
PE petroleum ether
PPh3 triphenylphosphine
Pr propyl
RB round bottom
rt or r.t. room temperature
RT retention time
SFC supercritical fluid chromatography
S03pyr sulfur trioxide pyridine complex - formula C5H5NS03
SPhos 2-dicyclohexyl phosphine-2',6'-dimethoxybiphenyl or
dicyclohexyl(2',6'-dimethoxy-[1 ,1 '-biphenyl]-2-yl)
t-BuOMe methyl t-butyl ether
T3P 1 -Propanephosphonic anhydride solution, 2,4,6-Tripropyl-1 ,3,5,2,4,6- trioxatriphosphorinane-2,4,6-trioxide
TFA trifluoroacetic acid
THF tetrahydrofuran
uv ultraviolet
[00239] Unless otherwise noted, all starting materials were obtained from commercial suppliers and used without further purification. Unless otherwise indicated, all temperatures are expressed in °C (degrees Centigrade). Unless otherwise indicated, all reactions are conducted under an inert atmosphere at ambient temperature.
[00240] All temperatures are given in degrees Celsius, all solvents are highest available purity and all reactions run under anhydrous conditions in an argon (Ar) or nitrogen (N2) atmosphere where necessary.
[00241] The following examples illustrate the invention. These examples are not intended to limit the scope of the present invention, but rather to provide guidance to the skilled artisan to prepare and use the compounds, compositions, and methods of the present invention.
[00242] While particular embodiments of the present invention are described, the skilled artisan will appreciate that various changes and modifications can be made without departing from the spirit and scope of the invention.
[00243] As used herein the symbols and conventions used in these processes, schemes and examples are consistent with those used in the contemporary scientific literature, for example, the Journal of the American Chemical Society or the Journal of Biological Chemistry. Unless otherwise noted, all starting materials were obtained from commercial suppliers and used without further purification.
[00244] All references to ether are to diethyl ether; brine refers to a saturated aqueous solution of NaCI. Unless otherwise indicated, all temperatures are expressed in °C (degrees Centigrade). All reactions are conducted under an inert atmosphere at room temperature unless otherwise noted, and all solvents are highest available purity unless otherwise indicated.
[00245] Equipment Description
[00246] 1H NMR spectra were recorded on a Bruker Ascend 400 spectrometer or a Varian 400 spectrometer. Chemical shifts are expressed in parts per million (ppm, δ units). Coupling constants are in units of hertz (Hz). Splitting patterns describe apparent multiplicities and are designated as s (singlet), d (doublet), t (triplet), q (quartet), quint (quintet), m (multiplet), br (broad).
[00247] Flash chromatography was run over Merck Silica gel 60 (230 - 400 mesh), or using a Teledyne Isco Combiflash Companion with normal phase, disposable Redi-Sep flash columns.
[00248] Mass spectra were run on an open access LC-MS system using electrospray ionization. The analytical low-resolution mass spectra (MS) were recorded on Waters SQD instrument with UPLC analysis was conducted on a Phenomenex Kinetex 1 .7um, 2.1 x 50mm XB-C18 column at 40CSQ using a gradient elution method. Solvent A: 0.2% formic acid (FA) in water; Solvent B: 0.15% FA in acetonitrile; 1 % - 99% Solvent B gradient over 1 .1 minutes and holding steady at 99% solvent B for another 0.4 minutes, at 1 ml/min flow rate. Alternatively, the analytical low-resolution mass spectra (MS) were recorded on Waters ACQUITY UPLC with SQ Detectors using a Waters BEH C18, 2.1 x 50 mm, 1 .7 μηι using a gradient elution method.
Solvent A: 0.1 % formic acid (FA) in water;
Solvent B: 0.1 % FA in acetonitrile;
30% B for 0.5 min followed by 30-100% B over 2.5 min.
Figure imgf000059_0001
[00250] Particular compounds disclosed herein may be made according to Scheme 1. For example, compound 1 can be prepared by reacting 4-bromo-1 ,2-dimethoxybenzene 9 with a ketone, as shown, using a palladium/xantphos Buchwald-Hartwig chemistry in sodium t-butoxide and THF. Reductive amination of 1 by reaction of compound 1 with sodium cyanoborohydride in ammonium acetate follows, to yield amine 2. Amine 2 is then reacted with formic acid in reflux conditions to yield amide 3. The dihydroisoquinoline compound 4 is obtained by a cyclization/condensation reaction using phosphoryl chloride in acetonitrile.
[00251] Reaction of dihydroisoquinoline 4 with ethyl 2-(ethoxymethylene)-3- oxobutanoate in ethanol under reflux conditions yields the tetrahydro-pyrido-isoquinoline- carboxylate 5 which is then aromatized with DDQ in DME under reflux conditions to yield the the dihydro-pyrido-isoquinoline-carboxylate 6. Ester hydrolysis of carboxylate 6 using NaOH in ethanol yields the isoquinoline carboxylic acid 7. [00252] Scheme 2 Preparation of Diastereomers
Figure imgf000060_0001
wherein R15, R15'' R16 and R16' are as described herein.
[00253] Alternatively, particular compounds disclosed herein may be made as shown in Scheme 2. For example, compound 10 can be prepared by reacting 4-bromo-1 ,2- dimethoxybenzene 9 with a ketone, as shown, using a palladium/xantphos Buchwald- Hartwig chemistry in sodium t-butoxide and THF. Reductive amination of 10 by reaction of compound 10 with sodium cyanoborohydride in ammonium acetate follows, to yield amine 11. Amine 11 is then reacted with formic acid in reflux conditions to yield amide 12. The dihydroisoquinoline compound 13 is obtained by a cyclization/condensation reaction using phosphoryl chloride in acetonitrile.
[00254] Reaction of dihydroisoquinoline 13 with ethyl 2-(ethoxymethylene)-3- oxobutanoate in ethanol under reflux conditions yields the tetrahydro-pyrido-isoquinoline- carboxylate 14 which is then aromatized with DDQ in DME under reflux conditions to yield the dihydro-pyrido-isoquinoline-carboxylate diastereomers 15A, 15B, 15C and 15D. Ester hydrolysis of diastereomers 15A, 15B, 15C and 15D using NaOH in ethanol yields the desired isoquinoline carboxylic acid as diastereomers 16A, 16B, 16C and 16D as minor and major products, respectively. [00255] Scheme 3
Figure imgf000061_0001
wherein R1 , R2, R3, R4, R5, R6, R7 amd R8 are as described herein.
[00256] Other particular compounds disclosed herein may be prepared as shown in Scheme 3, wherein substituted phenylmethanol 17 is reacted with thionyl chloride and pyridine in dichloromethane to yield the chloromethylbenzene 18. Reaction of 18 with a substituted ethyl isobutyrate in LDA yields the substituted phenylpropanoate 19. Reaction of substituted phenylpropanoate 19 with LiOH in THF, MeOH and H20 at 55 °C leads to the substituted phenylpropanoic acid 20. Compound 20 is then reacted with DPPA and triethylamine in t-BuOH to yield the substituted phenylpropanyl t-butyl carbamate 21.
Treatment of 21 with HCI in dioxane provides the phenylpropanyl amine hydrochloride 22, which upon treatment with formic acid and acetic anhydride in dioxane yields the substituted phenylpropanyl formamide 23. Reaction of 23 with POCI3 in acetonitrile yields the substituted dihydroisoquinoline 24. Upon addition of the substituted dihydroisoquinoline 24 with (E)-ethyl 2-(ethoxymethylene)-3-oxobutanoate in ethanol, the substituted oxo- tetrahydropyrido isoquinoline carboxylate compound 25 is formed. The dehydrogenation of carboxylate compound 25 with p-chloroanil in DME provides the substituted oxo- dihydropyrido isoquinoline carboxylate compound 26. Carboxylate compound 26
undergoes ester hydrolysis in the presence of LiOH in MeOH and H20 to yield the desired substituted oxo-dihydro isoquinoline carboxylic acid 27.
Figure imgf000062_0001
[00258] Compound Examples
[00259] Example 1
[00260] Compound 141 may be made as described below, and as shown in Scheme
4.
[00261] Compound 141 : 9'-(Benzyloxy)-10'-bromo-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000062_0002
[00262] Step 1 : Benzyl 3-(benzyloxy)-4-bromobenzoate
Figure imgf000062_0003
[00263] To a mixture of 4-bromo-3-hydroxybenzoic acid (90 g, 420 mmol) and K2C03 (173 g, 1256 mmol) in MeCN (1800 mL) was added benzyl bromide (142 g, 837 mmol) dropwise at room temperature over 30 min. The reaction mixture was stirred at 65°C overnight. The mixture was cooled to room temperature and water (900 mL) was added. The mixture was extracted 2 times with ethyl acetate (1800 mL). The combined organic layers were washed with bine (900 mL), dried over MgS04 (500 g), and concentrated under vacuo. The crude product was crystallized with MeOH (1800 mL) for 2 h at 0°C. The white solid was filtered and was washed with cold MeOH (100 ml_) to afford benzyl 3-(benzyloxy)- 4-bromobenzoate (140 g, yield: 85%). LCMS (ESI) m/z: 397.1 (M + 1 )+.
[00264] Step 2:
Figure imgf000063_0001
[00265] To a suspension of LiAIH4 (13.8 g, 363.6 mmol) in THF (130 mL) at -20°C was added a solution of benzyl 3-(benzyloxy)-4-bromobenzoate (144 g, 363.6 mmol) in THF (1500 mL) dropwise over 2 h. The mixture was stirred at -20°C for 2 h. Water (13.8 g) was added dropwise over 30 min at 0°C. To the mixture was added 40% aqueous NaOH (13.8 g) dropwise over 30 min and then stirred for another 30 min. The solid was filtered off and the filtrate concentrated. The residue was dissolved in ethyl acatate (1500 mL), washed with brine, dried 0ver MgSO4, and concentrated. The crude product was crystallized with petroleum ether (1500 mL) for 2 h at 0°C. The white solid was collected by filtration and washed with petroleum ether(500 mL) to afford (3-(benzyloxy)-4-bromophenyl)methanol (72 g, 68% yield). 1 H NMR (400 MHz, DMSO-de) δ 7.51 (dd, J = 12.9, 7.6 Hz, 3H), 7.42 (dd, J = 10.1 , 4.7 Hz, 2H), 7.36 - 7.32 (m, 1 H), 7.17 (d, J = 1 .4 Hz, 1 H), 6.90 - 6.80 (m, 1 H), 5.29 (t, J = 5.7 Hz, 1 H), 5.19 (s, 2H), 4.47 (t, J = 5.7 Hz, 2H).
[00266] Step 3:
Figure imgf000063_0002
[00267] To a solution of (3-(benzyloxy)-4-bromophenyl)methanol (62 g, 212 mmol) in DCM (620 mL) was added DMAP (1 .3 g, 10.6 mmol) and DIPEA (82.2 g, 637 mmol). A solution of TsCI (49 g, 254.8 mmol) in DCM (500 mL) was added dropwise at 0°C and then stirred at room temperature for 2 hrs. The reaction mixture was concentrated under reduced pressure to give the crude product, which was purified by flash chromatography (silica gel, 0 - 50% EtOAc in petroleum ether) to afford 3-(benzyloxy)-4-bromobenzyl 4- methylbenzenesulfonate (90 g, 95% yield). LCMS (ESI) m/z: 445.0 (M-1)".
[00268] Step 4:
Figure imgf000063_0003
[00269] To a solution of lithium bis(trimethylsilyl)amide solution (182 mL, 0.363 mol) in THF (500 mL) was added dropwise a solution of ethyl cyclobutanecarboxylate (38.7g, 0.302 mol) while keeping the temperature between -60°C and -65°C. After the addition the mixture was stirred at -60°C to -65°C for 2 hours. A solution of 3-(benzyloxy)-4-bromobenzyl 4-methylbenzenesulfonate (90 g, 0.201 mol) in THF (500 mL) was added dropwise and the resulting mixture stirred at -60°C to -65°C for 1 hour and then at room temperature for 3 hours. The reaction was quenched by addition of saturated ammonium chloride aqueous solution (200 mL) and the organic phase separated. The aqueous layer was extracted with ethyl acetate (1800 mL).The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with petroleum ethenethyl acetate (100:1 ) to afford the title compound as a yellow liquid (60 g, 80%). Ή NMR (400 MHz, DMSO-d6) δ 7.49 - 7.45 (m, 3H), 7.41 (dd, J = 8.0, 2.8 Hz, 2H), 7.33 (dd, J = 5.9, 3.6 Hz, 1 H), 6.96 (d, J = 1 .7 Hz, 1 H), 6.64 (dd, J = 8.1 , 1 .8 Hz, 1 H), 5.17 (s, 2H), 4.03 (d, J = 7.1 Hz, 2H), 3.00 (s, 2H), 2.28 - 2.20 (m, 2H), 1 .99 (ddd, J = 5.7, 4.1 , 2.1 Hz, 2H), 1 .87 - 1 .78 (m, 1 H), 1 .67 - 1 .56 (m, 1 H), 1 .14 (t, J = 7.1 Hz, 3H).
[00270] Step 5: romobenzyl)cvclobutane-1 -carboxylic acid
Figure imgf000064_0001
[00271] To a solution of ethyl 1 -(3-(benzyloxy)-4-bromobenzyl)cyclobutane-1 - carboxylate (60.0 g, 160.0 mmol) in EtOH (600 mL) and water (100 mL) was added LiOH (19.0 g, 802 mmol). The mixture was stirred for 4 hrs., acidified with 1 N HCI, and extracted with EtOAc (600 mL). The organic layer was washed with brine, dried over Na2S04, and concentrated to give the crude product which was purified by flash chromatography (silica gel, 0-50% EtOAc in petroleum ether) to afford 1 -(3-(benzyloxy)-4- bromobenzyl)cyclobutane-1 -carboxylic acid (45 g, 75% yield) as a white powder. LCMS (ESI) m/z: 373.1 (M-1)".
[00272] Step 6: bromobenzyl)cvclobutan-1 -amine
Figure imgf000064_0002
[00273] To a solution of 1 -(3-(benzyloxy)-4-bromobenzyl)cyclobutane-1 -carboxylic acid (45.2 g, 120mmol) in toluene was added TEA (15.0 g, 144mmol), followed by DPPA (40.0 g, 144mmol). The mixture was stirred at 80°C for 2 hrs. The mixture was cooled to room temperature and concentrated HCI (45 mL) was added. The new mixture was heated at 60°C for 2 hrs., cooled to 0°C, and basified with 1 N NaOH aqueous. The organic layer was washed with brine and dried over MgS04. The solution of 1 -(3-(benzyloxy)-4- bromobenzyl)cyclobutan-1 -amine in toluene was used to next step without purification. LCMS (ESI) m/z: 346.1 (M + 1)+. [00274] Step 7: N-(1 -(3-(Benzyloxy)-4-bromobenzyl)cvclobutyl)formamide
Figure imgf000065_0001
[00275] To a solution of 1 -(3-(benzyloxy)-4-bromobenzyl)cyclobutan-1 -amine (120.0 mmol) in toluene was added acetic formic anhydride (10.5 g, 120 mmol) dropwise at room temperature and the mixture stirred for 30 min. The mixture was concentrated under vacuo to give crude N-(1 -(3-(benzyloxy)-4-bromobenzyl)cyclobutyl)formamide. LCMS (ESI) m/z: 374.1 (M + 1)+.
[00276] Step 8: '-(Benzyloxy)-7'-bromo-4'H-spiro[cvclobutane-1 ,3'-isoquinoline
Figure imgf000065_0002
[00277] N-(1 -(3-(Benzyloxy)-4-bromobenzyl)cyclobutyl)formamide (120 mmol) was dissolved in MeCN (500 ml_). Phosphorus oxychloride (21 .7 g,144 mmol) was added dropwise over 30 min and then stirred overnight. The reaction mixture was cooled to 0°C and basified with ammonium hydroxide to pH = 9-10. The mixture was extracted 2 times with DCM. The combined organic layers were washed with brine and concentrated to give crude 6'-(Benzyloxy)-7'-bromo-4'H-spiro[cyclobutane-1 ,3'-isoquinoline (42.6 g, yield: 100%), which was used to next step without purification. LCMS (ESI) m/z: 356.1 (M + 1)+.
[00278] Step 9: Ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-1 '.2'.7'.1 1 b1- tetrahydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000065_0003
[00279] 6'-(Benzyloxy)-7'-bromo-4'H-spiro[cyclobutane-1 ,3'-isoquinoline (42.6 g, 120 mmol) was dissolved in EtOH (500 ml_) before ethyl-2-(methoxymethylene)-3- oxobutanoate was added. The mixture was heated at 85°C overnight. The mixture was cooled to 0°C and stirred for 2 hrs. The solid was filtered and washed with MTBE to afford ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-1 ',2',7',1 1 b'-tetrahydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylate (18 g, yield = 30% for 5 steps). LCMS (ESI) m/z: 496.1 (M + 1)+.
[00280] Step 10: Ethyl 9'-(benzyloxy)-10'-bromo^'-oxo^'J1- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000066_0001
[00281] To a suspension of ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-1 ',2',7',1 1 b'- tetrahydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (18 g, 36 mmol) in DME (270 mL) was added tetrach!oro-p-benzoquinone (7.1 g, 29 mmol) at room temperature. The mixture was heated at 75°C overnight. Solids were collected by filtration and washed with DME to afford ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate(17.0 g, yield: 94%). LCMS (ESI) m/z: 494.1 (M + 1)+.
[00282] Step 1 1 : 9'-(Benzyloxy)-10'-bromo-2'-oxo-2',7'-dihvdrospiro[cvclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000066_0002
[00283] To a solution of ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (20 mg, 0.04 mmol) in THF/EtOH/H20 (2 mL /1 mL /0.5 mL) was added LiOH (10.2 mg, 0.24 mmol). The mixture was stirred at r.t for 1 hr, acidified with 1 N HCI, and extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered, and concentrated to give crude product which was purified by reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford 9'-(Benzyloxy)-10'-bromo-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (5.6 mg, 29.8% yield ) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 10.15 (br, 1 H), 8.80 (s, 1 H), 8.32 (s, 1 H), 7.52 (d, J = 7.3 Hz, 2H), 7.44 (t, J = 7.4 Hz, 2H), 7.38(d, J = 7.8 Hz, 3H), 5.32 (s, 2H), 3.36 (s, 2H), 2.70-2.63 (m,2H), 2.07-2.01 (m, 2H), 1 .91 - 1 .85 (m, 2H). LCMS (ESI) m/z: 466.1 / 468.1 (M/M+2)+.
[00284] Example 2
[00285] 9'-(Cvclopropylmethoxy)-10'-(1 .1 -difluoroethyl)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000067_0001
[00286] Step 1 : Ethyl 10'-bromo-9'-hvdroxy-2'-oxo-2',7'-dihvdrospiro[cvclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000067_0002
[00287] A solution of ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (900 mg, 1 .82 mmol) in TFA (6 ml_) was heated at 65°C for 30 minutes and concentrated. The residue was dissolved in DCM, washed with sat. NaHC03, dried over Na2S04, and concentrated to give crude ethyl 10'-bromo-9'-hydroxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (770 mg crude, quant.). LCMS (ESI) m/z: 404.1 / 406.1 (M/M+2)+.
[00288] Step 2: Ethyl 10'-bromo-9'-(cvclopropylmethoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000067_0003
[00289] To the solution of ethyl 10'-bromo-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (770 mg, 1 .9 mmol) in MeCN (5 ml_) was added K2C03 (787 mg, 5.7 mmol) and (bromomethyl)cyclopropane (282 mg, 2.09 mmol). The mixture was heated at 65°C for 4 hours. The mixture was cooled to room temperature and partitioned between EtOAc and H20. The phases were separated and the aqueous layer extracted 2 times with EtOAc, The combined organic layers were washed brine and concentrated. The residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM to afford ethyl 10'-bromo-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (800 mg, 96% yield over two steps). LCMS (ESI) m/z: 458.2 / 460.2 (M/M+2)+. [00290] Step 3: Ethyl 10'-acetyl-9'-(cvclopropylmethoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000068_0001
[00291] To a solution of ethyl 10'-bromo-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (280 mg, 0.61 mmol) in DMF (4 mL) was added 1 -(vinyloxy)butane (122 mg, 1 .22 mmol), 1 ,3- bis(diphenylphosphino)propane (30 mg, 0.07 mmol), Pd(OAc)2 (13 mg, 0.06 mmol), and TEA (123 mg, 1 .22 mmol). The mixture was heated at 90°C overnight. The mixture was cooled to room temperature, treated with 1 N HCI, and extracted with EA. The organic phase was washed with brine, dried over Na2S04, and concentrated. The residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM to afford ethyl 10'-acetyl- 9'-(cyclopropylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate. (140 mg, 54% yield). LC MS (ESI) m/z: 422.3 (M + 1)+.
[00292] Step 4: Ethyl 9'-(cvclopropylmethoxy)-10'-(2-methyl-1 .3-dithiolan-2-yl)-2'- oxo-2', 7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000068_0002
[00293] To a solution of ethyl 10'-acetyl-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (80 mg, 0.19 mmol) in DCM (2 mL) at 0°C was added ethane-1 ,2-dithiol (89 mg, 0.95 mmol), followed by boron trifluoride etherate (81 mg, 0.57 mmol) dropwise. The resulting mixture was stirred at 0°C for 4 hours and then partitioned between DCM and ice water. The layers were separated, and the aqueous layer was extracted with DCM. The combined organic layers were dried over Na2S04, filtered, and concentrated to give crude ethyl 9'-(cyclopropylmethoxy)-10'-(2- methyl-1 ,3-dithiolan-2-yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (94 mg). LCMS (ESI) m/z: 498.3 (M + 1)+. [00294] Step 5: Ethyl 9'-(cvclopropylmethoxy)-1 Q'-(1 .1 -difluoroethyl)-2'-oxo-2'.7'- dihydrospirofcyclobuta '-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000069_0001
[00295] To a solution of ethyl 9'-(cyclopropylmethoxy)-10'-(2-methyl-1 ,3- dithiolan-2-yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (94 mg, 0.19 mmol) in DCM (5 mL) was added NIS (86 mg, 0.38 mmol), followed by dropwise addition of pyridine hydrofluoride (75 mg, 0.76 mmol). The reaction mixture was stirred for 5 minutes, quenched with ice water, and extracted with DCM. The organic layer was dried over Na2S04, filtered, and concentrated. The residue was purified by prep.TLC (10% MeOH in DCM) to afford ethyl 9'-(cyclopropylmethoxy)-10'-(1 ,1 - difluoroethyl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (30 mg, 36% yield over two steps). LCMS (ESI) m/z: 444.3 (M + 1)+.
[00296] Step 6: 9'-(Cvclopropylmethoxy)-10'-(1 .1 -difluoroethyl)-2'-oxo-2'.7'- dihydrospirofcyclobuta '-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000069_0002
[00297] To a solution of ethyl 9'-(cyclopropylmethoxy)-10'-(1 ,1 -difluoroethyl)-
2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (30 mg, 0.068 mmol) in THF (2 mL) was added LiOH (1 1 mg, 0.27 mmol) in water (0.5 mL). The mixture was stirred at room temperature for 20 minutes and then acidified with 1 N HCI to pH = 6. The mixture was extracted 2 times with 15% IPA in DCM. The combined organic layers were dried over Na2S04, filtered, and concentrated. The residue was purified by reverse phase HPLC (Gilson, C18, 20%-100% MeCN in water with 0.1 % formic acid) to afford 9'- (cyclopropylmethoxy)-10'-(1 ,1 -difluoroethyl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid . (5 mg, 33% yield) as a white solid. LCMS (ESI) m/z: 416.9 (M + 1)+. Ή NMR (400 MHz, DMSO-d6) δ 8.81 (s, 1 H), 7.96 (s, 1 H), 7.30 (s, 1 H), 7.26 (s, 1 H), 4.06 (d, J = 6.9 Hz, 2H), 3.39 (s, 2H), 2.70 - 2.63 (m, 2H), 2.1 1 - 2.00 (m, 5H), 1 .93 - 1 .86 (m, 2H), 1 .32 - 1 .27 (m, 1 H), 0.63 - 0.59 (m, 2H), 0.40 - 0.36 (m, 2H).
[00298] Example 3 [00299] 9'-(CvclopropylmethoxyV10'-ethyl-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000070_0001
[00300] Step 1 : Ethyl 9'-(cvclopropylmethoxyV2'-oxo-10'-vinyl-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000070_0002
[00301] To a solution of ethyl 10'-bromo-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (200 mg, 0.44 mmol) and 4,4,5,5-tetramethyl-2-vinyl-1 ,3,2-dioxaborolane (136 mg, 0.88mmol) was added Pd(PPh3)4 (50 mg, 0.044 mmol) and Cs2C03 (429 mg, 1 .32 mmol) in dioxane (3 mL) and water (1 drop). The mixture was stirred at 85 °C under N2 for 2 hrs. The mixture was allowed to cool to room temperature and water was (1 mL) added. The mixture was extracted with 2 times ethyl acetate. The combined organic layers were washed with brine and concentrated. The residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM to afford ethyl 9'-(cyclopropylmethoxy)-2'-oxo-10'-vinyl-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (130 mg, 73% yield). LCMS (ESI) m/z: 406.3 (M + 1)+.
[00302] Step 2: Ethyl 9'-(cvclopropylmethoxy)-10'-ethyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000070_0003
[00303] To a solution of ethyl 9'-(cyclopropylmethoxy)-2'-oxo-10'-vinyl-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (130 mg, 0.32 mmol) in MeOH (5 mL) was added Pd (20 mg, on activated carbon(10%)(wetted with ca. 55% Water). The mixture was stirred at room temperature under H2 at balloon pressure overnight. The catalyst was filtered off. The filtrate was concentrated to give crude ethyl 9'- (cyclopropylmethoxy)-10'-ethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (100 mg, 78% yield). LCMS (ESI) m/z: 408.3 (M + 1)+.
[00304] Step 3: 9'-(Cvclopropylmethoxy)-10'-ethyl-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000071_0001
[00305] To a solution of ethyl 9'-(cyclopropylmethoxy)-10'-ethyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (100 mg, 0.25 mmol) in MeOH (5 ml_) and H20 (1 ml_) was added LiOH (18 mg, 0.74mmol). The reaction mixture was stirred at room temperature for 30 minutes. The reaction was acidified with 1 N HCI to pH = 6 and extracted with 15% IPA in DCM. The combined organic layers were dried over Na2S04, filtered, and concentrated. The residue was purified by reverse phase HPLC (Gilson, C18, 20%-100% MeCN in water with 0.1 % formic acid) to afford 9'- (cyclopropylmethoxy)-10'-ethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid (50 mg, 53% yield) as a white solid. LCMS (ESI) m/z: 380.35 (M + 1)+. Ή NMR (400 MHz, DMSO-d6)
[00306] δ 8.79 (s, 1 H), 7.81 (s, 1 H), 7.29 (s, 1 H), 7.02 (s, 1 H), 3.97 (d, J = 6.8 Hz, 2H), 3.31 (s, 2H), 2.68 - 2.59 (m, 4H), 2.05 - 1.98 (m, 2H), 1.93 - 1 .84 (m, 2H), 1.32 - 1.26 (m, 1 H), 1.20 (t, J = 7.5 Hz, 3H), 0.64 - 0.57 (m, 2H), 0.40 - 0.33 (m, 2H).
[00307] Example 4
[00308] 9'-(Cvclopropylmethoxy)-10'-(difluoromethyl)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000071_0002
[00309] Step 1 : Ethyl 9'-(cvclopropylmethoxy)-2'-oxo-10'-vinyl-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000071_0003
[00310] A mixture of ethyl 10'-bromo-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (1 .5 g, 3.3 mmol), potassium vinyltrifluoroborate (876 mg, 6.54 mmol), Pd(PPh3)4 (381 mg, 0.33 mmol), and Na2C03 (1 .04 g, 9.81 mmol) in EtOH (30 mL) was stirred at 90°C under N2 overnight. The mixture was concentrated and the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to give ethyl 9'-(cyclopropylmethoxy)-2'-oxo-10'-vinyl-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (1 g, 75.8% yield) as a yellow solid. LCMS (ESI) m/z: 406.2 (M + 1 ) +.
[00311] Step 2: Ethyl 9'-(cvclopropylmethoxy)-10'-formyl-2'-oxo-2'.7'- dihydrospirofcyclo
[00312] butane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000072_0001
[00313] To a stirred solution of ethyl 9'-(cyclopropylmethoxy)-2'-oxo-10'-vinyl-
2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (300 mg, 0.74 mmol) in THF (10 mL) and water (2.5 mL) was added K2OSO4 2H2O (27 mg, 0.074 mmol) at 0°C. The mixture was stirred at 0°C for 15 min. before Nal04 (316.6 mg,1 .48 mmol) was added. The mixture was stirred at room temperature overnight. Ethyl acetate and water were added. The organic layer was separated, dried over Na2S04, and concentrated. The residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to ethyl 9'- (cyclopropylmethoxy)-10'-formyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate
[00314] (160 mg, 53% yield) as a yellow solid. LCMS (ESI) m/z: 408.3 (M + 1 )+ .
[00315] Step 3: Ethyl 9'-(cvclopropylmethoxy)-10'-(difluoromethyl)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000072_0002
[00316] To a solution of ethyl 9'-(cyclopropylmethoxy)-10'-formyl-2'-oxo-2',7'- dihydro spiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (60 mg, 0.147 mmol) in DCM (8 mL) was added a solution of DAST (59.4 mg, 0.368 mmol) in DCM (2 mL) dropwise at 0°C. The mixture was stirred at room temperature overnight. The mixture was poured into saturated NaHC03 at 0°C slowly. The organic layer was separated, dried over Na2S04, concentrated, and the residue was purified by flash chromatography (silica gel, 0- 20% MeOH in DCM) to give ethyl 9'-(cyclopropylmethoxy)-10'-(difluoromethyl)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (17 mg, 27% yield) as a yellow oil. LCMS (ESI) m/z: 430.21 (M + 1)+.
[00317] Step 4: 9'-(Cvclopropylmethoxy)-10'-(difluoromethyl)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000073_0001
[00318] To a solution of ethyl 9'-(cyclopropylmethoxy)-10'-(difluoromethyl)-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (17 mg, 0.04 mmol) in THF/EtOH/H20 (2 mL /1 mL /0.5 mL) was added LiOH (10 mg, 0.24 mmol). After stirred at r.t for 1 hr, the resulting mixture was neutralized with 1 N HCI and extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated. The residue was purified by reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford 9'-(cyclopropylmethoxy)-10'-(difluoromethyl)-2'-oxo- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (11.8 mg, 74.2% yield) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 10.15 (br, 1 H), 8.81 (s, 1 H), 8.12 (s, 1 H), 7.33 (s, 1 H), 7.28 (s, 1 H),7.22-6.94(m, 1 H), 4.06 (d, J = 6.9 Hz, 2H), 3.41 (s, 2H), 2.70 - 2.63 (m, 2H), 2.03 - 1.99 (m, 2H), 1.92 - 1.86 (m, 2H),1.31-1 .28(m, 1 H),0.63 - 0.58 (m, 2H), 0.41-0.37 (m, 2H). LCMS (ESI) m/z: 402.3 (M + 1)+.
[00319] Example 5
[00320] 9'-(CvclopropylmethoxyV10'-(fluoromethvn-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000073_0002
[00321] Step 1 : Ethyl 9'-(cvclopropylmethoxy)-10'-(hvdroxymethyl)-2'-oxo-2',7'- dihydrospiro [cyclobuta '-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000074_0001
[00322] To a stirred solution of ethyl 9'-(cyclopropylmethoxy)-10'-formyl-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (160 mg, 0.39 mmol) in THF (2 mL) and DCM (2 mL) at 0°C was added NaBH4 (15 mg , 0.39 mmol) slowly. The mixture was stirred at 0°C for 30 min, quenched with sat. NhUCI, and extracted with DCM. The organic layer was washed with brine, dried over Na2S04, filtered, concentrated, and the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to give ethyl9'-(cyclopropylmethoxy)-10'-(hydroxymethyl)-2'-oxo-2',7'- dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (80 mg, 49.8% yield) as a yellow solid. LCMS (ESI) m/z: 410.2 (M + 1 )+.
[00323] Step 2: Ethyl 9'-(cvclopropylmethoxyV10'-(fluoromethyr)-2'-oxo-2'.7'- dihydrospiro [cyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000074_0002
[00324] To a solution of ethyl 9'-(cyclopropylmethoxy)-10'-(hydroxymethyl)-2'- oxo-2', 7'-dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (40 mg, 0.098 mmol) in DCM (2 mL) was added a solution of DAST (15.8 mg, 0.098 mmol) in DCM (1 mL) dropwise at 0 °C and the mixture stirred at room temperature overnight. The mixture was poured into saturated NaHCCb at 0 °C slowly. The organic layer was separated, dried over Na2S04, concentrated and the residue purified by flash chromatography (silica gel, 0- 20% MeOH in DCM) to give ethyl 9'-(cyclopropylmethoxy)-10'-(fluoromethyl)-2'-oxo-2',7'- dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (23 mg, 57% yield) as a yellow oil. LCMS (ESI) m/z: 412.3 (M + 1)+.
[00325] Step 3: 9'-(Cvclopropylmethoxy)-10'-(fluoromethyl)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000075_0001
[00326] To a solution of ethyl 9'-(cyclopropylmethoxy)-10'-(fluoromethyl)-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (23 mg, 0.056 mmol) in THF/EtOH/H20 (2 mL /1 mL /0.5 mL) was added LiOH (14 mg, 0.34 mmol). The mixture was stirred at r.t for 1 hr, acidified with 1 N HCI, and extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered, concentrated, and the residue purified by reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford 9'-(cyclopropylmethoxy)-10'-(fluoromethyl)-2'-oxo-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylic acid
(16.1 mg, 75.2% yield ) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 12.15 (br, 1 H), 8.80 (s, 1 H), 8.11 (s, 1 H), 7.28 (s, 1 H), 7.17 (s, 1 H), 5.44 (d, J = 47.9 Hz, 2H), 4.02(d, J = 6.9 Hz, 2H),3.38 (s, 2H), 2.70-2.62 (m,2H), 2.06-1.99 (m, 2H), 1.94-1.86(m, 2H),1.32- 1.26(m, 1 H),0.63-0.57(m, 2H),0.41-0.35(m, 2H). For 1H NMR, found 22 out of 22 protons. LCMS (ESI) m/z: 384.3 (M + 1)+.
[00327] Example 6
[00328] 9'-(Cvclopropylmethoxy)-2'-oxo-10'-(trifluoromethyl)-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000075_0002
[00329] Step 1 : Ethyl 9'-(cvclopropylmethoxyV2'-oxo-10'-(trifluoromethvn-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000075_0003
[00330] To a solution of ethyl 10'-bromo-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (70 mg, 0.15 mmol) in DMF (2 mL) was added Cul (71 mg, 0.38 mmol) and methyl 2,2-difluoro-2- (fluorosulfonyl)acetate (286 mg, 1.5 mmol). The mixture was heated at 85 °C for 28 h. The mixture was heated 120 °C and stirred overnight at 120 °C. The mixture was cooled to room temperature. Water was added and then extracted 2 times with ethyl acetate. The combined organic layers were washed with brine and concentrated. The residue was purified by TLC (DCM: MeOH = 15: 1 ) to afford ethyl 9'-(cyclopropylmethoxy)-2'-oxo-10'- (trifluoromethyl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (10 mg, 6% yield). LCMS (ESI) m/z: 448.6 (M + 1)+.
[00331] Step 2: 9'-(Cvclopropylmethoxy)-2'-oxo-10'-(trifluoromethyl)-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000076_0001
[00332] To a solution of ethyl 9'-(cyclopropylmethoxy)-2'-oxo-10'-
(trifluoromethyl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (10 mg, 0.03 mmol) in EtOH (5 ml_) and H20 (1 ml_) at room temperature was added LiOH (2 mg, 0.1 mmol). The mixture was heated at 65 °C for 30 min. The mixture was acidified to pH = ~1 and purified by reverse phase HPLC (C18, 0-30% Acetonitrile in the H20 with 0.1 % formic acid) to afford 9'-(cyclopropylmethoxy)-2'-oxo-10'-(trifluoromethyl)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (1 .5 mg, 18% yield) as a white solid. LCMS (ESI) m/z: 420.3 (M + 1)+. 1 H NMR (400 MHz, DMSO-d6) δ 8.81 (s, 1 H), 8.22 (s, 1 H), 7.46 (s, 1 H), 7.39 (s, 1 H), 4.12 (d, J = 6.9 Hz, 2H), 3.43 (s, 2H), 2.69 - 2.62 (m, 2H), 2.05 - 2.00 (m, J = 6.8 Hz, 2H), 1 .92 -1 .86 (m, 2H), 1 .31 - 1 .27 (m, 1 H), ,0.62 - 0.57 (m, 2H), 0.41 - 0.37 (m, 2H).
[00333] Example 7
[00334] 10'-Methoxy-9'-(3-methoxypropoxy)-2'-oxo-2.2'.3.5.6.7'- hexahvdrospiro[pyran-4,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000076_0002
[00336] n-Butyllithium (2.5 M in hexanes) (0.581 mL, 1 .45 mmol) was added dropwise to a -78 °C solution of diisopropylamine (0.207 mL, 1 .45 mmol) in tetrahydrofuran (THF) (5 mL). The mixture was allowed to warm to room temperature and stirred for 15 minutes. The mixture was cooled to -78 °C before methyl tetrahydro-2H-pyran-4- carboxylate (0.185 mL, 1 .38 mmol) was added dropwise. The mixture was warmed to 0 °C and stirred for 15 minutes. A white precipitate formed. A solution of 4-(bromomethyl)-1 - methoxy-2-(3-methoxypropoxy)benzene (200 mg, 0.692 mmol) in tetrahydrofuran (THF) (1 .7 mL) was added dropwise. The mixture was stirred an additional 20 minutes at 0 °C and then quenched with saturated ammonium chloride. The mixture was extracted 2 times with ethyl acetate. The combined organic layers were washed with brine, dried over sodium suflate, and concentrated to give crude methyl 4-(4-methoxy-3-(3- methoxypropoxy)benzyl)tetrahydro-2H-pyran-4-carboxylate as a clear oil. LCMS (ESI) m/z: 353.3 (M+1)+.
[00337] Step 2: 4-(4-Methoxy-3-(3-methoxypropoxy)benzyl)tetrahvdro-2H-pyran-4- carboxylic acid
Figure imgf000077_0001
[00338] A solution of lithium hydroxide monohydrate (291 mg, 6.92 mmol) in water (2 mL) was added to a solution of methyl 4-(4-methoxy-3-(3- methoxypropoxy)benzyl)tetrahydro-2H-pyran-4-carboxylate (244 mg, 0.692 mmol) in 1 ,4- dioxane (2 mL) and the mixture heated at 85 °C overnight. The mixture was allowed to cool to room temperature, quenched with 1 M hydrochloric acid, and extracted 3 times with 2- methyltetrahydrofuran. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give crude 4-(4-methoxy-3-(3- methoxypropoxy)benzyl)tetrahydro-2H-pyran-4-carboxylic acid (234 mg). N62802-31 -1 . LCMS was consistent with desired product. LCMS (ESI) m/z: 339.3 (M+1)+.
[00339] Step 3: 4-(4-Methoxy-3-(3-methoxypropoxy)benzyl)tetrahvdro-2H-pyran-4- amine
Figure imgf000077_0002
[00340] Potassium tert-butoxide (160 mg, 1 .430 mmol) was added to a mixture of 4-(4-methoxy-3-(3-methoxypropoxy)benzyl)tetrahydro-2H-pyran-4-carboxylic acid (242 mg, 0.715 mmol) in 1 ,4-dioxane (5 mL) and tert-butanol (2 mL). Diphenyl
phosphorazidate (0.308 mL, 1 .43 mmol) was added dropwise and the mixture heated to 85 °C. A thick white precipitate formed very quickly. The mixture was allowed to cool to room temperature and concentrated. The residue was mixed with 1 ,4-dioxane (5 mL) and 5N hydrochloric acid (10 mL, 50.0 mmol). The mixture was heated at 100 °C for 2 hours, cooled, and poured into ice with excess concentrated ammonium hydroxide. The mixture was extracted 2 times with 2-methyltetrahydrofuran and the combined organic layers were concentrated. The residue was purified by reverse phase medium pressure
chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient). Fractions were concentrated to give 4-(4-methoxy-3-(3-methoxypropoxy)benzyl)tetrahydro- 2H-pyran-4-amine (102 mg, 0.330 mmol, 46% yield). LCMS (ESI) m/z: 310.3 (M+1)+.
[00341] Step 4: N-(4-(4-Methoxy-3-(3-methoxypropoxy)benzyl)tetrahvdro-2H-pyran- 4-yl)formamide
Figure imgf000078_0001
[00342] A solution of 4-(4-methoxy-3-(3-methoxypropoxy)benzyl)tetrahydro-2H- pyran-4-amine (102 mg, 0.330 mmol) and formic acid (0.126 mL, 3.30 mmol) in 1 ,4-dioxane (2 mL) were heated at 100 °C overnight. The mixture was allowed to cool and was concentrated to give crude N-(4-(4-methoxy-3-(3-methoxypropoxy)benzyl)tetrahydro-2H- pyran-4-yl)formamide (1 1 1 mg, 0.329 mmol, 100 % yield). LCMS (ESI) m/z: 338.4 (M+1)+.
[00343] Step 5: 7-Methoxy-6-(3-methoxypropoxy)-2',3',5',6'-tetrahydro-4H-
Figure imgf000078_0002
[00344] N-(4-(4-Methoxy-3-(3-methoxypropoxy)benzyl)tetrahydro-2H-pyran-
4-yl)formamide (1 1 1 mg, 0.329 mmol) in acetonitrile (15 mL) were cooled in an ice bath before phosphorus oxychloride (0.037 mL, 0.395 mmol) was added dropwise with stirring. The mixture was heated at 60 °C for 4 hours and concentrated. The residue was dissolved in ethyl acetate and cooled in an ice bath before water and 30% NH4OH were added to bring the pH = ~1 1 . The organic layer was collected and the aqueous layer extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give 7-methoxy-6-(3-methoxypropoxy)-2',3',5',6'-tetrahydro-4H- spiro[isoquinoline-3,4'-pyran] (71 mg, 0.222 mmol, 67.6 % yield). LCMS (ESI) m/z: 320.3 (M+1)+. [00345] Step 6: Ethyl 10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2.2'.3.5.6.7'- hexahvdrospiro[pyran-4,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000079_0001
[00346] 7-Methoxy-6-(3-methoxypropoxy)-2',3',5',6'-tetrahydro-4H- spiro[isoquinoline-3,4'-pyran] (71 mg, 0.222 mmol) and ethyl 2-(ethoxymethylene)-3- oxobutanoate (0.193 mL, 1 .1 1 mmol) in ethanol (1 mL) were heated at reflux overnight. The mixture was cooled, concentrated, and the residue dissolved in 1 ,2-dimethoxyethane (DME) (1 mL). p-Chloranil (54.7 mg, 0.222 mmol) was added and the mixture heated at 85 °C for 2 hours. The mixture was concentrated and the residue purified by silica chromatography eluting with a gradient of 0% to 100% ethanol in ethyl acetate. Fractions were concentrated to give ethyl 10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2,2',3,5,6,7'-hexahydrospiro[pyran- 4,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (49 mg, 0.107 mmol, 48% yield) as a white powder. LCMS (ESI) m/z: 458.3 (M+1)+.
[00347] Step 7: 10'-Methoxy-9'-(3-methoxypropoxy)-2'-oxo-2.2'.3.5.6.7'- hexahvdrospiro[pyran-4,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000079_0002
[00348] A solution of lithium hydroxide monohydrate (41 .3 mg, 0.984 mmol) in water (1 .667 mL) was added to a solution of ethyl 10'-methoxy-9'-(3-methoxypropoxy)-2'- oxo-2,2',3,5,6,7'-hexahydrospiro[pyran-4,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (45 mg, 0.098 mmol) in methanol (5 mL) and the mixture heated at 60 °C for 2 hours. The mixture was allowed to cool to room temperature, diluted with water, acidified with 1 M hydrochloric acid, and extracted 3 times with dichloromethane. The combined organic layers were washed with brine and concentrated. The residue was purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient). Fractions were concentrated. The residue was lyophililzed (acetonitrile / water) to give 10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2,2',3,5,6,7'-hexahydrospiro[pyran-4,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (30 mg, 0.070 mmol, 71 % yield) as a white solid. Ή NMR (400 MHz, DMSO-c/6) δ ppm 8.77 (s, 1 H), 7.53 (s, 1 H), 7.48 (s, 1 H), 7.14 (s, 1 H), 4.10 (t, J=6.64 Hz, 2 H), 3.88 (s, 3 H), 3.78 - 3.85 (m, 2 H), 3.64 - 3.73 (m, 2 H), 3.48 (t, J=6.25 Hz, 2 H), 3.41 (s, 2 H), 3.25 (s, 3 H), 2.1 1 - 2.26 (m, 2 H), 2.00 (quin, J=6.25 Hz, 2 H), 1 .76 (d, J=12.10 Hz, 2 H). LCMS (ESI) m/z: 430.3 (M+1)+.
[00349] Example 8
[00350] 10'-Chloro-9'-(cvclopropylmethoxy)-2'-oxo-2',7'-dihvdrospiro[cyclohexane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000080_0001
[00351] Step 1 : Methyl 1 -(4-chloro-3-methoxybenzyl)cvclohexanecarboxylate
Figure imgf000080_0002
[00352] n-Butyllithium (2.5 M in hexanes) (6.06 mL, 15.2 mmol) was added dropwise to a -78 °C solution of diisopropylamine (2.16 mL, 15.2 mmol) in tetrahydrofuran (THF) (40 mL). The mixture was allowed to warm to room temperature and stirred for 15 minutes. The mixture was cooled to -78 °C before methyl cyclohexanecarboxylate (2.06 mL, 14.4 mmol) was added dropwise. The mixture was warmed to 0 °C and stirred for 15 minutes. A solution of 4-(bromomethyl)-1 -chloro-2-methoxybenzene (1 .7 g, 7.22 mmol) in tetrahydrofuran (THF) (13.3 mL) was added dropwise. The mixture was stirred an additional 20 minutes at 0 °C and then quenched with saturated ammonium chloride. The mixture was extracted 2 times with ethyl acetate. The combined organic layers were washed with brine, dried over sodium suflate, and concentrated to give crude methyl 1 -(4- chloro-3-methoxybenzyl)cyclohexane-1 -carboxylate (2.14 g). 1H NMR (400 MHz,
CHLOROFORM-d) δ ppm 7.15 - 7.28 (m, 1 H), 6.51 - 6.62 (m, 2 H), 3.84 (s, 3 H), 3.60 (s, 3 H), 2.74 (s, 2 H), 1 .36 - 2.09 (m, 4 H), 1 .12 - 1 .34 (m, 6 H).
[00353] Step 2: -(4-Chloro-3-methoxybenzyl)cvclohexanecarboxylic acid
Figure imgf000080_0003
[00354] A solution of lithium hydroxide monohydrate (3.41 g, 81 mmol) in water (10 mL) was added to a solution of methyl 1 -(4-chloro-3-methoxybenzyl)cyclohexane- 1 -carboxylate (2.41 g, 8.12 mmol) in 1 ,4-dioxane (10 mL) and the mixture heated at 100 °C overnight. The mixture was allowed to cool to room temperature, quenched with 1 M hydrochloric acid, and extracted 3 times with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give crude 1 -(4- chloro-3-methoxybenzyl)cyclohexane-1 -carboxylic acid (2.29 g). LCMS (ESI) m/z: 281 .1 (M- 1)-.
[00355] Step 3: 1 -(4-Chloro-3-methoxybenzyl)cvclohexanamine
Figure imgf000081_0001
[00356] Diphenyl phosphorazidate (3.84 ml_, 17.82 mmol) was added dropwise to a 0 °C stirring mixture of 1 -(4-chloro-3-methoxybenzyl)cyclohexane-1 -carboxylic acid (2.29 g, 8.10 mmol) and triethylamine (2.483 ml_, 17.82 mmol) in toluene (40 ml_). The mixture was allowed to warm to room temperature and stirred for 15 minutes before being heated at 80 °C for 1 hour. The mixture was allowed to cool to room temperature before a mixture 5N hydrogen chloride (40 ml_, 200 mmol) and 1 ,4-dioxane (40 ml_) was added. The mixture was heated at 80 °C with vigorous stirring for 1 hour. The mixture was allowed to cool to room temperature and was extracted with ethyl ether. The ethyl ether layer was back-extracted 2 times with 1 N hydrochloric acid. The combined acidic aqueous layers were basified with 1 M sodium hydroxide and extracted 3 times with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give 1 -(4-chloro-3-methoxybenzyl)cyclohexan-1 -amine (1 .1 g, 4.3 mmol, 54 % yield) as an oil. LCMS (ESI) m/z: 254.2 (M+1 )+.
[00357] Step 4: N-(1 -(4-chloro-3-methoxybenzyl)cvclohexyl)formamide
Figure imgf000081_0002
[00358] A solution of 1 -(4-chloro-3-methoxybenzyl)cyclohexan-1 -amine (1 .1 g, 4.3 mmol) and formic acid (1 .663 mL, 43.3 mmol) in 1 ,4-dioxane (25 mL) was heated at 100 °C for 3 days. The mixture was allowed to cool and was concentrated to give crude N- (1 -(4-chloro-3-methoxybenzyl)cyclohexyl)formamide (1 .22 g). LCMS (ESI) m/z: 282.2 (M+1)+.
[00359] Step 5: 7'-Chloro-6'-methoxy-4'H-spiro[cvclohexane-1 ,3'-isoquinolinel
Figure imgf000081_0003
[00360] N-(1 -(4-Chloro-3-methoxybenzyl)cyclohexyl)formamide (1 .2 g, 4.26 mmol) in acetonitrile (40 mL) was cooled in an ice bath before phosphorus oxychloride (0.476 mL, 5.1 1 mmol) was added dropwise with stirring. The mixture was heated at 60 °C for 2 hours and concentrated. The residue was dissolved in ethyl acetate and cooled in an ice bath before water and 30% NH4OH were added to bring the pH = ~1 1 . The organic layer was collected and the aqueous layer extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give crude 7'-chloro-6'-methoxy-4'H-spiro[cyclohexane-1 ,3'-isoquinoline] (1 .12 g). LCMS (ESI) m/z: 264.2 (M+1)+.
[00361] Step 6: Ethyl 10'-chloro-9'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclohexane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000082_0001
[00362] 7'-Chloro-6'-methoxy-4'H-spiro[cyclohexane-1 ,3'-isoquinoline] (1 .12 g, 4.25 mmol) and ethyl 2-(ethoxymethylene)-3-oxobutanoate (3.69 mL, 21 .2 mmol) in ethanol (20 mL) were heated at reflux overnight. The mixture was cooled, concentrated, and the residue dissolved in 1 ,2-dimethoxyethane (DME) (20 mL). p-Chloranil (1 .04 g, 4.25 mmol) was added and the mixture heated at 85 °C for 2 hours. The mixture was allowed to cool to room temperature and then cooled to 0 °C in an ice bath. The precipitate was collected by filtration, washed with cold 1 ,2-dimethoxyethane, and dried to give ethyl 10'- chloro-9'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (560 mg, 1 .39 mmol, 33 % yield) as solid with a pale light green tint. LCMS (ESI) m/z: 402.3 (M+1)+.
[00363] Step 7: 10'-Chloro-9'-hvdroxy-2'-oxo-2',7'-dihvdrospiro[cvclohexane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000082_0002
[00364] Boron tribromide (1 M in dichloromethane) (2.84 mL, 2.84 mmol) was added dropwise to a 0 °C solution of ethyl 10'-chloro-9'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (380 mg, 0.946 mmol) in dichloromethane (DCM) (5 mL). The mixture was allowed to warm to room temperature and stirred overnight. Additional boron tribromide was added (1 eq) and the mixture stirred overnight. The mixture was poured into ice, diluted with 1 M hydrochloric acid, and extracted 3 times with ethyl acetate. The combined organic layers were concentrated. The residue was dissolved in methanol (5 mL), basified with 1 M sodium hydroxide (10 ml_), and heated at 70 C for 3 hours. The mixture was allowed to cool to room temperature, acidified with 1 N hydrochloric acid, and extracted 3 times with ethyl acetate. The combined organic layers were dried over sodium sulfate and concentrated to give 10'-chloro-9'-hydroxy-2'-oxo-2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid (266 mg, 0.739 mmol, 78 % yield). LCMS (ESI) m/z: 360.2 (M+1)+.
[00365] Step 8: 10'-Chloro-9'-(cvclopropylmethoxy)-2'-oxo-2'.7'- dihydrospirofcyclohexa -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000083_0001
[00366] 10'-Chloro-9'-hydroxy-2'-oxo-2',7'-dihydrospiro[cyclohexane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (122 mg, 0.339 mmol), potassium carbonate (469 mg, 3.39 mmol), and (bromomethyl)cyclopropane (0.263 ml_, 2.71 mmol) in N,N- dimethylformamide (DMF) (1 ml_) were stirred at room temperature overnight. The mixture was quenched with water and extracted 2 times with ethyl acetate. The combined organic layers were washed with 5% aqueous lithium chloride, washed with brine, and concentrated to give crude cyclopropylmethyl 10'-chloro-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate. The residue was dissolved in methanol (1 ml_) before an aqueous solution of lithium hydroxide monohydrate (285 mg, 6.78 mmol) was added. The mixture was heated at 60 °C for 3 hours. The mixture was allowed to cool to room temperature, acidified with 1 N hydrochloric acid, and a precipitate collected by filtration. The product was slurried in acetonitrile, sonicated, and the precipitate collected by filtration. The sample was air dried. The crude product was purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient). Fractions were lyophilized to give 10'-chloro-9'- (cyclopropylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylic acid (47 mg, 0.1 1 mmol, 33 % yield). 1H NMR (400 MHz, DMSO-cf6) δ ppm 8.79 (s, 1 H), 8.15 (s, 1 H), 7.38 (s, 1 H), 7.29 (s, 1 H), 3.91 - 4.12 (m, 2 H), 1 .84 - 2.00 (m, 2 H), 1 .73 (d, J=1 1 .72 Hz, 2 H), 1 .62 (br. s., 3 H), 1 .46 - 1 .58 (m, 2 H), 1 .17 - 1 .39 (m, 3 H), 0.50 - 0.66 (m, 3 H), 0.35 (q, J=4.69 Hz, 2 H). LCMS (ESI) m/z: 414.3 (M+1)+. [00367] Example 9
[00368] 9'-(CvclopropylmethoxyV10'-methyl-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000084_0001
[00369] Step 1 : Ethyl 9'-(cvclopropylmethoxy)-10'-methyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000084_0002
[00370] Ethyl 10'-bromo-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (200 mg, 0.44 mmol), methylboronic acid (105 mg, 1 .75 mmol), Pd(PPh3)4 (254 mg, 0.22 mmol), Cs2C03 (571 mg, 1 .75 mmol), toluene (40 mL) and water (2 mL) under nitrogen were stirred for 3 h at 100 °C. The mixture was cooled to room temperature, added to water (40 mL), and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and
concentrated. The residue was purified by column chromatography eluting with
hexane/ethyl acetate. Fractions were concentrated to afford ethyl 9'-(cyclopropylmethoxy)- 10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (1 12mg, 65% yield). LCMS (ESI) m/z: 394.2 (M + 1)+.
[00371] Step 2: 9'-(CvclopropylmethoxyV10'-methyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000084_0003
[00372] To a solution of ethyl 9'-(cyclopropylmethoxy)-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.25 mmol) in EtOH (5 mL) and H20 (1 mL) at room temperature was added LiOH (24 mg, 1 mmol). The mixture was heated at 65 °C for 30 min. and cooled to room temperature. The mixture was acidified to pH = ~4 and purified by reverse phase HPLC (C18, 0-30% acetonitrile in the H20 with 0.1 % formic acid) to afford 9'-(cyclopropylmethoxy)-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (55 mg, 60% yield) as a white solid. LCMS (ESI) m/z: 366.2 (M + 1)+. Ή NMR (400 MHz, DMSO-d6) δ 8.78 (s, 1 H), 7.86 (s, 1 H), 7.25 (s, 1 H), 7.01 (s, 1 H), 3.96 (d, J = 6.9 Hz, 2H), 3.30 (s, 2H), 2.67 - 2.59 (m,2H), 2.21 (s, 3H), 2.05 - 1 .98 (m, 2H), 1 .92 - 1 .84 (m, 2H), 1 .31 - 1 .25 (m, 1 H), 0.63 - 0.58 (m, 2H), 0.39 - 0.35 (m, 2H).
[00373] Example 10
[00374] 10'-Methyl-2'-oxo-9'-(pyrrolidin-1 -yl)-2',7'-dihvdrospiro[cvclobutane-1 ,6'- pyrido[2,1 -alisoquinolin '-carboxylic acid
Figure imgf000085_0001
[00375] Step 1 : Ethyl 9'-(benzyloxy)-10'-methyl^'-oxo^'J1- dihvdrospiro[cvclobutan '-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000085_0002
[00376] Ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'-dihydrospiro[cyclobutane-
1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (5.0 g, 10 mmol), methylboronic acid (2.4 g, 40 mmol), Pd(Ph3)4 (5.8 g, 5 mmol), Cs2C03 (13 g, 40 mmol), toluene (40 mL) and water (2 mL) were stirred overnight at 100 °C under nitrogen. The reaction mixture cooled to room temperature and water (100 mL) added. The mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica chromatography eluting with DCM/MeOH. Fractions were concentrated to afford ethyl 9'-(benzyloxy)-10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (3.4 g, 79% yield), LCMS (ESI) m/z: 430.32 (M + 1)+. [00377] Step 2: Ethyl 9'-hvdroxy-10'-methyl-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000086_0001
[00378] A solution of ethyl 9'-(benzyloxy)-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (3.4 g, 8 mmol) in TFA (40 ml_) was stirred at 65 °C. After the reaction was finished, the mixture was concentrated. The residue was dissolved in DCM, washed with saturated NaHC03 aqueous, dried over MgS04 and concentrated. The residue was purified by flash chromatography to afford ethyl 9'-hydroxy-10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (2.15 g, 75% yield). LCMS (ESI) m/z: 340.12 (M + 1)+.
[00379] Step 3: Ethyl 10'-methyl-2'-oxo-9'-(((trifluoromethyl)sulfonyl)oxy)-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000086_0002
[00380] To a solution of ethyl 9'-hydroxy-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (2.0 g, 5.9 mmol) in CH3CN was added PhN(Tf)2 (2.5 g, 7.1 mmol), DIPEA (2.3 g, 17.7 mmol), and DMAP (36 mg, 0.295 mmol). The mixture was stirred at room temperature for 2 h, filtered, and the residue was washed with EtOAc. The filtrate was concentrated and the residue purified by flash chromatography (silica gel, MeOH/DCM 1 : 10) to afford ethyl 10'-methyl-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate (1 .3 g, 47% yield). LCMS (ESI) m/z: 472.2 (M + 1)+.
[00381] Step 4: Ethyl 10'-methyl^'-oxo-g'-foyrrolidin-l -yl)-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000086_0003
[00382] A Schlenk tube was charged with Pd(dba)2 (120.75 mg, 0.2 mmol),
BINAP (130.76 mg, 0.21 mmol), Cs2C03 (137 mg, 0.42 mmol) and toluene (2 ml_). To this mixture was added a solution of ethyl 10'-methyl-2'-oxo-9'-(((trifluoromethyl)sulfonyl)oxy)- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.21 mmol) and pyrrolidine (18 mg, 0.25 mmol) in toluene (1 ml_) via syringe. The mixture was heated at 80 °C with stirring until the starting material had been consumed as judged by LCMS analysis. The mixture was cooled to room temperature, diluted with ethyl acetate, filtered, and concentrated. The crude material was purified by flash chromatography on silica gel to afford ethyl 10'-methyl-2'-oxo-9'-(pyrrolidin-1 -yl)-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (49 mg, 60% yield). LCMS (ESI) m/z: 393.2 (M + 1)+.
[00383] Step 5: 10'-Methyl-2'-oxo-9'-(pyrrolidin-1 -yl)-2',7'-dihvdrospiro[cyclobutane-
Figure imgf000087_0001
[00384] To a solution of ethyl 10'-methyl-2'-oxo-9'-(pyrrolidin-1 -yl)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (20 mg, 0.05 mmol) in MeOH (2 mL) and water (0.5 mL) was added LiOH (1 .2 mg, 0.05 mmol). The reaction mixture was stirred at room temperature for 1 h and then purified by reverse phase HPLC (C18, 0-30% MeCN in H20 with 0.1 % formic acid) to afford 10'-methyl-2'-oxo-9'-(pyrrolidin- 1 -yl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (9 mg, 50% yield) as a yellow solid. LCMS (ESI) m/z: 365.9 (M + 1 )+. Ή NMR (400 MHz, DMSO- de) δ 8.75 (s, 1 H), 7.68 (s, 1 H), 7.15 (s, 1 H), 6.70 (s, 1 H), 3.41 -3.38 (m, 4H), 3.24 (s, 2H), 2.64-2.57 (m, 2H), 2.38 (s, 3H), 2.04-2.00 (m, 2H), 1 .93-1 .88 (m, 6H).
[00385] Example 11
[00386] 10'-Methyl-9'-(oxetan-3-ylmethoxy)-2'-oxo-2',7'-dihvdrospiro[cyclobutane-
Figure imgf000087_0002
[00387] Step 1 : Ethyl 10'-methyl-9'-(oxetan-3-ylmethoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000088_0001
[00388] A Schlenk was charged with ethyl 9'-hydroxy-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.29 mmol), oxetan-3-ylmethyl 4-methylbenzenesulfonate (43 mg, 0.35 mmol) and CH3CN (2 ml_).
Potassium carbonate (48 mg, 0.35 mmol) was added and the mixture stirred for 6 h at 85 °C. After cooling, the obtained inorganic salt was filtered off. Ethyl acetate was added to the filtrate and washed with brine. After drying with anhydrous magnesium sulfate the solvent was evaporated and the residue purified by column chromatography to afford ethyl 10'- methyl-9'-(oxetan-3-ylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (65 mg, 55% yield). LCMS (ESI) m/z: 410.2 (M + 1)+ .
[00389] Step 2: 10'-Methyl-9'-(oxetan-3-ylmethoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000088_0002
[00390] To a solution of ethyl 10'-methyl-9'-(oxetan-3-ylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (50 mg, 0.12 mmol) in MeOH (2 ml_) and water (0.5 ml_) was added LiOH (14.4 mg, 0.6 mmol). The mixture was stirred at room temperature for 1 h, acidified to pH = ~4 and purified by reverse phase HPLC (C18, 0-40% MeCN in H20 with 0.1 % formic acid) to afford 10'-methyl-9'-(oxetan-3- ylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (22 mg, 48% yield) as a white solid. LCMS (ESI) m/z: 382.8 (M + 1)+.Ή NMR (400 MHz, DMSO-de) δ 8.79 (s, 1 H), 7.88 (s, 1 H), 7.26 (s, 1 H), 7.09 (s, 1 H), 4.74 (dd, J = 7.8, 6.1 Hz, 2H), 4.48 (t, J = 6.0 Hz, 2H), 4.31 (d, J =6.3 Hz, 2H), 3.47 - 3.42 (m, 1 H), 3.30 (s, 2H), 2.68 - 2.61 (m, 2H), 2.20 (s, 3H), 2.06 - 2.00 (m, 2H), 1 .94 - 1 .86 (m, 2H).
[00391] Example 12 [00392] 9'-(CvclopentylmethoxyV10'-methyl-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000089_0001
[00393] Step 1 : Ethyl 9'-(cvclopentylmethoxy)-10'-methyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000089_0002
[00394] To a solution of ethyl 9'-hydroxy-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (90 mg, 0.265 mmol) in DMF (5 ml_) was added K2C03 (73.2 mg, 0.530 mmol) and (bromomethyl)cyclopentane (51 .8 mg, 0.318 mmol). The mixture was heated at 90 °C for 3 hours. The mixture was cooled to room temperature and partitioned between EtOAc and H20. The organic layer was separated and the aqueous layer was extracted 2 times with EtOAc. The combined organic layers were washed brine, filtered, concentrated, and the residue purified by flashed chromatography (silica gel, 0-20 % MeOH in DCM to afford ethyl 9'-(cyclopentylmethoxy)- 10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (60 mg, 53.7% yield). LCMS (ESI) m/z: 422.4 (M + 1 )+.
[00395] Step 2: 9'-(Cvclopentylmethoxy)-10'-methyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000089_0003
[00396] To a solution of ethyl 9'-(cyclopentylmethoxy)-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (60 mg, 0.142 mmol) in MeOH (5 ml), NaOH (45.5 mg, 1 .14 mmol) dissolved in H20 (1 ml) was added. The mixture was stirred at room temperature for 2 h, acidified to pH = ~5 with 1 N HCI, and diluted with EtOAc (20ml_) and H20 (30 ml_). The mixture was extracted with EtOAc (50 mL x 2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by revers phase HPLC (C18 column, 30%-100% MeCN in H20, with 0.1 % formic acid) to give 9'-(cyclopentylmethoxy)-10'-methyl-2'-oxo-2',7'-dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylic acid (16 mg, 28.6% yield) as a white solid. LCMS (ESI) m/z: 394.3 (M + 1)+. Ή NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1 H), 7.85 (s, 1 H), 7.25 (s, 1 H), 7.04 (s, 1 H), 3.97 (d, J = 6.8 Hz, 2H), 3.32 (s, 2H), 2.68 - 2.60 (m, 2H), 2.39 - 2.34 (m, 1 H), 2.20 (s, 3H), 2.04 - 1.99 (m, 2H), 1.93 - 1 .86 (m, 2H), 1.83 - 1.77 (m, 2H), 1.65 - 1 .54 (m, 4H), 1.42 - 1.36 (m, 2H).
[00397] Example 13
[00398] 10'-Methoxy-9'-f 1 -methyl-1 H-pyrazol^-vD^'-oxo^'J'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000090_0001
[00399] Stepl : Ethyl 10'-methoxy-9'-f 1 -methyl-1 H-pyrazol^-vD^'-oxo^'J'- dihydrospiro
[00400] [cyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000090_0002
[00401] A mixture of ethyl 10'-methoxy-2'-oxo-9'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate (80 mg,0.164 mmol), 1-methyl-4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)-1 H-pyrazole(61.5 mg, 0.295 mmol), Pd(dppf)CI2 (13 mg, 0.0164mmol), and CH3COOK (48 mg, 0.492 mmol) in 1 ,4-dioxane(1 ml_) and H20 (1 drop) was stirred at 100°C under N2 overnight. The mixture was concentrated and the residue purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to afford ethyl 10'-methoxy-9'-(1-methyl-1 H-pyrazol-4-yl)- 2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (70 mg, 100% yield) as a yellow solid. LCMS (ESI) m/z: 420.31 (M + 1)+.
[00402] Step 2: 10'-Methoxy-9'-(1 -methyl-1 H-pyrazol-4-yl)-2'-oxo-2'.7'-dihvdrospiro
[00403] [cyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000091_0001
[00404] To a solution of ethyl 10'-methoxy-9'-(6-methoxypyridin-3-yl)-2'-oxo-
2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (70 mg, 0.167 mmol) in THF/EtOH/H20(2 mL /1 mL /0.5 mL) was added LiOH (42 mg, 1 mmol). The mixture was stirred at r.t for 1 hr, acidified with 1 N HCI, and extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered, concentrated, and the residue purified by reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford 10'-methoxy-9'-(1 -methyl-1 H-pyrazol-4-yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylic acid (13.2 mg, 20.3% yield ) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 1 1.94 (br, 1 H), 8.83 (s, 1 H), 8.25 (s, 1 H), 8.01 (s, 1 H), 7.73 (s, 1 H), 7.58 (s, 1 H), 7.56 (s, 1 H), 4.02 (s, 3H), 3.90 (s, 3H), 3.32 (s, 2H),2.69-2.62 (m, 2H), 2.07-2.01 (m, 2H), 1 .93-1.86 (m, 2H). LCMS (ESI) m/z: 392.3(M + 1)+.
[00405] Example 14
[00406] 9'-(Furan-3-yl)-10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000091_0002
[00407] Stepl : Ethyl 9'-(furan-3-yl)-10'-methoxy^'-oxo^'J1- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000091_0003
[00408] To a solution of ethyl 10'-methoxy-2'-oxo-9'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate (100 mg, 0.205 mmol) in DMF(5 ml) was added furan-3-ylboronic acid (47 mg, 0.41 mmol), Pd(dppf)CI2 (20 mg,0.02mmol), K2C03 (55 mg, 0.41 mmol), and H20(1 ml). The mixture was stirred at 100 °C for 3 h, cooled to room temperature, and diluted with EtOAc (20ml_) and H20 (30 ml_). The mixture was extracted with EtOAc (50 mL x 2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel, 0-10% MeOH in DCM) to afford ethyl 9'- (furan-3-yl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (68 mg, 81 .8% yield) as a yellow solid. LCMS (ESI) m/z: 406.5 (M + 1 )+.
[00409] Step 2: 9'-(Furan-3-yl)-10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000092_0001
[00410] To a solution of ethyl 9'-(furan-3-yl)-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (68 mg, 0.168 mmol) in MeOH(10 ml) was added NaOH (53.7 mg, 1 .34 mmol) in H20 (1 ml_). The resulting mixture was stirred at 40 °C for 2 h. The mixture was acidified to pH = ~5 with 1 N HCI. The mixture was diluted with EtOAc (20ml_) and H20 (30 mL), and then extracted with EtOAc (50 mL x 2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by Gilson (C18 column, 40%-100% MeCN in H20,with 0.1 % formic acid) to give 9'-(furan-3-yl)-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (1 1 .5 mg, 18% yield) as a white solid. LCMS (ESI) m/z: 378.4 (M + 1)+. Ή NMR (400 MHz, DMSO-d6) δ 8.84 (s, 1 H), 8.28 (s, 1 H), 7.79 (s, 1 H), 7.75 (s, 1 H), 7.62 (s, 1 H), 7.59 (s, 1 H), 7.10 (s, 1 H), 4.04 (s, 3H), 3.34 (s, 2H), 2.71 - 2.62 (m, 2H), 2.07 - 2.00 (m, J = 6.7 Hz, 2H), 1 .96 - 1 .86 (m, J = 15.4, 8.0 Hz, 2H).
[00411] Example 15
[00412] 10'-Chloro-9'-methoxy-2'-oxo-2',7'-dihvdrospiro[cvclohexane-1 ,6'-pyrido[2,1 - alisoquinolinel-3'-carboxylic acid
Figure imgf000092_0002
[00413] Step 1 : 10'-Chloro-9'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclohexane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000093_0001
[00414] A solution of lithium hydroxide monohydrate (39.7 mg, 0.946 mmol) in water (0.667 mL) was added to a solution of ethyl 10'-chloro-9'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (38 mg, 0.095 mmol) in methanol (1 mL) and the mixture heated at 60 °C for 2 hours. The mixture was allowed to cool to room temperature, diluted with water, acidified with 1 M hydrochloric acid, and extracted 3 times with dichloromethane. The combined organic layers were washed with brine and concentrated to ~3 mL of dichloromethane. Hexanes were added and the precipitate collected by filtration. Drying under vacuum gave 10'-chloro-9'-methoxy-2'-oxo- 2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (15 mg, 0.040 mmol, 42 % yield) as a white solid. Ή NMR (400 MHz, DMSO-c/6) δ ppm 8.77 (s, 1 H), 8.15 (s, 1 H), 7.37 (s, 1 H), 7.30 (s, 1 H), 3.90 (s, 3 H), 3.28 (s, 2 H), 1 .84 - 2.00 (m, 2 H), 1 .72 (d, J=1 1 .71 Hz, 2 H), 1 .43 - 1 .66 (m, 5 H), 1 .18 - 1 .35 (m, 1 H). LCMS (ESI) m/z: 374.3 (M+1)+.
[00415] Example 16
[00416] 10'-(Difluoromethyl)-2'-oxo-9'-((tetrahvdro-2H-pyran-4-yl)methoxy)-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000093_0002
[00417] Step 1 : (Tetrahydro-2/-/-pyran-4-yl)methyl 4-methylbenzenesulfonate
Figure imgf000093_0003
[00418] To a 0°C solution of (tetrahydro-2H-pyran-4-yl)methanol (1 .1 g, 9.47 mmol) in anhydrous DCM was added DMAP (~3 mg, cat.) and TEA (1 .9 g, 18.94 mmol), followed by TsCI (1 .8 g, 9.5mmol). The mixture was stirred at room temperature overnight. The solvent was removed and the residue purified by flash chromatography (silica gel, 0- 40% ethyl acatate in petroleum ether) to afford tetrahydro-2H-pyran-4-yl)methyl 4- methylbenzenesulfonate (1 .2 g, yield,47%) as a white solid. LCMS (ESI) m/z: 271 .17 (M +
[00419] Step 2: Ethyl 9'-(benzyloxy)-2'-oxo-10'-vinyl-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000094_0001
[00420] To a solution of ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (6.0 g, 12mmol) in EtOH (120 mL) was added potassium vinyl trifluoroborate (3.2 g, 24 mmol), Pd(PPh3)4 (1 .4 g, 1 .2 mmol) and Na2C03 (3.85 g, 36.3 mmol) under N2 atmosphere. The mixture was heated at 80 °C overnight. After cooling to rt, the mixture was filtered and washed with DCM (100 mL). The filtrate was concentrated and the residue purified by column chromatography using MeOH in DCM (1/15) to afford the title compound (4.6 g, 86% yield). LCMS (ESI) m/z: 442.7 (M + 1)+.
[00421] Step 3: Ethyl 9'-(benzyloxy)-10'-formyl-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000094_0002
[00422] To a solution of ethyl 9'-(benzyloxy)-2'-oxo-10'-vinyl-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (4.6 g, 10mmol) in THF (100 mL) and H20 (25 mL) was added K20s04 2H20 (384 mg, 1 .04 mmol) at 0 °C. Sodium periodate (8.9 g, 41 .6 mmol) was added in portions. The mixture was allowed to warm to room temperature and stirred for 2 h. The mixture was filtered and washed with DCM (100 mL). The filtrate was concentrated and the residue purified by column chromatography eluting with MeOH in DCM (1 /15). Fractions were concentrated to afford ethyl 9'-(benzyloxy)-10'-formyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (4.6 g, 86% yield). LCMS (ESI) m/z: 444.3 (M + 1 )+. [00423] Step 4: Ethyl 9'-(benzyloxy)-10'-toifluoromethyl^'-oxo^'J1- dihvdrospiro[cvclobutan -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000095_0001
[00424] To a solution of ethyl 9'-(benzyloxy)-10'-formyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (1.3 g, 2.93 mmol) in DCM (40 mL) was added DAST (1.4 g, 8.79 mmol) dropwise at 0 °C under N2. The mixture was stirred at rt overnight, diluted with DCM (80 mL), washed with sat.NaHC03 (50 mL), washed with brine (50 mL), and concentrated. The residue was purified by column chromatography eluting with MeOH in DCM (1/20) to afford ethyl 9'-(benzyloxy)-10'- (difluoromethyl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'- carboxylate (750 mg, 76% yield). LCMS (ESI) m/z: 466.6 (M + 1)+.
[00425] Step 5: Ethyl 1 O'-toifluoromethviyg'-hvdroxy^'-oxo^'J1- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000095_0002
[00426] A mixture of ethyl 9'-(benzyloxy)-10'-(difluoromethyl)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (750 mg, 1.61 mmol) and added 10% Pd/C (75 mg) was stirred under H2 at (15 psi) In MeOH (20 mL) at rt for 5 hrs. The mixture was filtered and the filtrate concentrated. The residue was triturated with ethyl acetate to afford ethyl 10'-(difluoromethyl)-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (280 mg, 46% yield). LCMS (ESI) m/z: 376.4 (M + 1)+.
[00427] Step 6: Ethyl 10'-(difluoromethyl)-2'-oxo-9'-((tetrahvdro-2H-pyran-4- yl)methoxy)-2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000095_0003
[00428] To the solution of ethyl 10'-(difluoromethyl)-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.266 mmol) in DMF (3 mL) was added K2C03 (1 10 mg, 0.8 mmol) and (tetrahydro-2/-/-pyran-4- yl)methyl 4-methylbenzenesulfonate (108 mg, 0.4 mmol). The mixture was heated at 80°C for 5 hours. The mixture was cooled to room temperature and partitioned between EtOAc and H20. The organic layer was separated and the aqueous layer extracted 2 times with EtOAc. The combined organic layers were washed brine and concentrated. The residue was purified by flashed chromatography (silica gel, 0-30 % MeOH in DCM to afford ethyl 10'-(difluoromethyl)-2'-oxo-9'-((tetrahydro-2H-pyran-4-yl)methoxy)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (30 mg, 24% yield over two steps). LCMS (ESI) m/z: 474.3 (M + 1)+.
[00429] Step 7: 10'-(Difluoromethyl)-2'-oxo-9'-((tetrahydro-2H-pyran-4-yl)methoxy)- 2',7'-dihvdrospiro[cyclobutane- '-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000096_0001
[00430] To a solution of ethyl 10'-(difluoromethyl)-2'-oxo-9'-((tetrahydro-2H- pyran-4-yl)methoxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (30 mg, 0.06mmol) in THF (3 mL) was added LiOH (4 mg, 0.18 mmol) in water (1 mL). The mixture was stirred at room temperature for 20 minutes, acidified with 1 N HCI to pH = 6, and extracted 2 times with 15% IPA in DCM. The combined organic layers were dried over Na2S04 and concentrated. The residue was purified by reverse phase HPLC (Gilson, C18, 30%-100% MeCN in water with 0.1 % formic acid) to afford 10'- (difluoromethyl)-2'-oxo-9'-((tetrahydro-2/-/-pyran-4-yl)methoxy)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (7.6 mg, 28% yield) as a white solid. LCMS (ESI) m/z: 446.31 (M + 1 )+. Ή NMR (400 MHz, DMSO-d6) δ 8.81 (s, 1 H), 8.13 (s, 1 H), 7.33 (s, 1 H), 7.30 (s, 1 H), 7.08 (t, J = 54.7 Hz, 1 H), 4.05 (d, J = 6.3 Hz, 2H), 3.90 (dd, J = 1 1 .2, 3.1 Hz, 2H), 3.42 (s, 2H), 3.39 - 3.33 (m, 2H), 2.71 - 2.63 (m, 2H), 2.07 - 1 .98 (m, 3H), 1 .93 - 1 .86 (m, 2H), 1 .73 - 1 .68 (m, 2H), 1 .43 - 1 .34 (m, 2H).
[00431] Example 17 [00432] 10'-(DifluoromethylV2'-oxo-9'-((tetrahvdrofuran-3-vnmethoxyV2'.7'-
Figure imgf000097_0001
[00434] A solution of ethyl 9'-(benzyloxy)-10'-formyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (3.9 g, 8.8 mmol) in TFA (80 mL) was heated at 70 °C for 4 h. The solvent was removed under reduced pressure. The mixture was diluted with DCM (150 mL) and washed with brine (80 mL x 3). The organic layer was dried over Na2S04, concentrated, and the residue triturated with ethyl acetate / petroleum ether to afford ethyl 10'-formyl-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (2.4 g, 77% yield). LCMS (ESI) m/z: 354.5 (M + 1)+.
[00435] Step 2:
Figure imgf000097_0002
[00436] To a solution of (tetrahydrofuran-3-yl)methanol (1 .0 g, 9.8 mmol) in
DCM (15 mL) was added N,N-dimethylpyridin-4-amine (122 mg, 1 mmol), TEA (2.0 g, 20 mmol) and 4-methylbenzenesulfonyl chloride (1 .86 g, 9.8 mmol). The reaction mixture was stirred at rt for 3 h. The mixture was concentrated and residue was purified by column chromatography using ethyl acetate in petroleum ether (1/4) to afford tetrahydrofuran-3- yl)methyl 4-methylbenzenesulfonate (900 mg, 36% yield). LCMS (ESI) m/z: 257.4 (M + 1)+. [00437] Step 3: Ethyl 10'-formyl-2'-oxo-9'-((tetrahvdrofuran-3-yl)methoxyV2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000098_0001
[00438] To a solution of ethyl 10'-formyl-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.28 mmol) in DMF (4 mL) was added (tetrahydrofuran-3-yl)methyl 4 methylbenzenesulfonate (144 mg, 0.56 mmol) and K2C03 (1 16 mg, 0.84 mmol). The mixture was heated at 90 °C overnight, poured into water (30 mL), and extracted with DCM (25 X 2 mL). The combined organic layers were washed with brine (30 X 4 mL), concentrated, and the residue purified by column chromatography eluting with MeOH in DCM (1/15) to afford ethyl 10'-formyl-2'-oxo- 9'-((tetrahydrofuran-3-yl)methoxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (102 mg, 84% yield). LCMS (ESI) m/z: 438.4 (M + 1)+.
[00439] Step 4: Ethyl 10'-(difluoromethyl)-2'-oxo-9'-((tetrahydrofuran-3-yl)methoxy)- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000098_0002
[00440] To a solution of ethyl 10'-formyl-2'-oxo-9'-((tetrahydrofuran-3- yl)methoxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (102 mg, 0.23 mmol) in DCM (4 mL) was added DAST (148 mg, 0.92 mmol) dropwise at 0 °C under N2. The mixture was stirred at rt overnight, diluted with DCM (30 mL), washed with sat.NaHC03 (20 mL), and washed with brine (20 mL). The organic phase was concentrated to afford ethyl 10'-(difluoromethyl)-2'-oxo-9'-((tetrahydrofuran-3-yl)methoxy)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (85 mg, 79% yield). LCMS (ESI) m/z: 460.4 (M + 1)+. [00441] Step 5: 10'-(DifluoromethylV2'-oxo-9'-((tetrahvdrofuran-3-yl)methoxyV2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000099_0001
[00442] To a solution of ethyl 10'-(difluoromethyl)-2'-oxo-9'-((tetrahydrofuran-
3-yl)methoxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate ( 85 mg, 0.18 mmol) in EtOH (6 mL) was added NaOH (36 mg, 0.9 mmol) in H20 (1 .5 ml_). The mixture was stirred at rt for 2 h and concentrated. The mixture was acidified to pH = ~1 by 1 N HCI and extracted with DCM (10 ml X 3). The combined organic layers were washed with brine (30 mL), dried over Na2S04, and concentrated. The residue was purified by reverse phase HPLC (C18, 30- 100 % acetonitrile in H20 with 0.1 % formic acid) to afford 10'-(difluoromethyl)-2'-oxo-9'-((tetrahydrofuran-3-yl)methoxy)-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (25 mg, 32% yield) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 8.81 (s, 1 H), 8.13 (s, 1 H), 7.33 (d, J = 7.3 Hz, 2H), 7.07 (t, J = 54.7 Hz, 1 H), 4.30 - 3.99 (m, 2H), 3.92 - 3.75 (m, 2H), 3.75 - 3.64 (m, 1 H), 3.63 - 3.53 (m, 1 H), 3.41 (s, 2H), 2.85 - 2.58 (m, 3H), 2.28 - 1 .96 (m, 3H), 2.00 - 1 .81 (m, 2H), 1 .80 - 1 .61 (m, 1 H). LCMS (ESI) m/z: 432.4 (M + 1)+.
[00443] Example 18
[00444] 10'-(Difluoromethyl)-9'-(3-hvdroxy-3-methylbutoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000099_0002
[00445] Step 1 : 3-Hvdroxy-3-methylbutyl 4-methylbenzenesulfonate
Figure imgf000099_0003
[00446] To a solution of 3-methylbutane-1 ,3-diol (1 .36 g, 13 mmol) in DCM
(20 mL) was added N,N-dimethylpyridin-4-amine (122 mg, 1 mmol), TEA (2.6 g, 26 mmol), and 4-methylbenzenesulfonyl chloride (2.5 g, 13 mmol). The reaction mixture was stirred at rt for 3 h, concentrated, and the residue was purified by column chromatography using ethyl acetate in petroleum ether (1/4) to afford 3-hydroxy-3-methylbutyl 4- methylbenzenesulfonate (1 .4 g, 42% yield). LCMS (ESI) m/z: 259.5 (M + 1)+.
[00447] Step 3: Ethyl 10'-(difluoromethyl)-9'-(3-hvdroxy-3-methylbutoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000100_0001
[00448] To a solution of ethyl 10'-formyl-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (50 mg, 0.13 mmol) in DMF (3 mL) was added 3-hydroxy-3-methylbutyl 4-methylbenzenesulfonate (36 mg, 0.26 mmol) and K2C03 (54 mg, 0.39 mmol). The mixture was heated at 90 °C for 4 h, cooled to room temperature, poured into water (30 mL), and extracted with DCM (25 X 2 mL). The combined organic layers were washed with brine (30 X 4 mL), concentrated, and the residue purified by Prep-TLC eluting with MeOH in DCM (1/20) to afford ethyl 10'- (difluoromethyl)-9'-(3-hydroxy-3-methylbutoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylate (40 mg, 65% yield). LCMS (ESI) m/z: 462.5 (M +
[00449] Step 3: 10'-(Difluoromethvn-9'-(3-hvdroxy-3-methylbutoxyV2'-oxo-2'.7'- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000100_0002
[00450] To a solution of ethyl 10'-(difluoromethyl)-9'-(3-hydroxy-3- methylbutoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (4 mg, 0.09 mmol) in MeOH (4 mL) was added LiOH H20 (1 1 mg, 0.26 mmol) in H20 (1 mL). The mixture was stirred at rt for 3 h, concentrated, and acidified to pH = ~1 with 1 N HCI. The mixture was extracted with DCM (10 mL X 3). The combined organic layers were washed with brine (20 mL),dried over Na2S04, and concentrated. The residue was purified by reverse phase HPLC (C18, 30- 60 % acetonitrile in H20 with 0.1 % formic acid) to afford 10'-(difluoromethyl)-9'-(3-hydroxy-3-methylbutoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (16 mg, 42% yield) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1 H), 8.12 (s, 1 H), 7.33 (s, 2H), 7.05 (t, J = 54.8 Hz, 1 H), 4.44 (s, 1 H), 4.29 (t, J = 8 Hz, 2H), 3.43 (s, 2H), 2.69-2.65 (m,2H), 2.07-2.00 (m, 2H), 1 .93-1 .89 (m, 4H), 1 .19 (s, 6H). LCMS (ESI) m/z: 434.3 (M + [00451] Example 19
[00452] 10'-(Difluoromethyl)-9'-(oxetan-3-ylmethoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000101_0001
[00453] Step 1 : Oxetan-3-ylmethyl 4-methylbenzenesulfonate
Figure imgf000101_0002
[00454] To a solution of oxetan-3-ylmethanol (750 mg, 8.5 mmol) in DCM (15 mL) was added N,N-dimethylpyridin-4-amine (122 mg, 1 mmol), TEA (2.0 g, 20 mmol) and 4-methylbenzenesulfonyl chloride (1 .6 g, 8.5 mmol). The mixture was stirred at rt for 3 h, concentrated, and the residue was purified by column chromatography using ethyl acatate in petroleum ether (1 /4) to afford oxetan-3-ylmethyl 4-methylbenzenesulfonate (1 .4 g, 70% yield). LCMS (ESI) m/z: 243.3 (M + 1 )+.
[00455] Step 2: Ethyl 10'-formyl-9'-hvdroxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane-
Figure imgf000101_0003
[00456] A solution of ethyl 9'-(benzyloxy)-10'-formyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (3.9 g, 8.8 mmol) in TFA (80 mL) was heated at 70 °C for 4 h. The solvent was removed under reduced pressure. The mixture was diluted with DCM (150 mL), washed with brine (80 mL x 3), dried over Na2S04, concentrated, and the residue triturated with EA/PE to afford ethyl 10'-formyl- 9'-hydroxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (2.4 g, 77% yield). LCMS (ESI) m/z: 354.5 (M + 1)+. [00457] Step 3: Ethyl 10'-formyl-9'-(oxetan-3-ylmethoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000102_0001
[00458] To a solution of ethyl 10'-formyl-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.28 mmol) in DMF (4 mL) was added oxetan-3-ylmethyl 4-methylbenzenesulfonate (136 mg, 0.56 mmol) and K2C03 (1 16 mg, 0.84 mmol). The reaction mixture was heated at 90 °C for 4 h, cooled to room temperature, poured into water (30 mL), and extracted with DCM (25 X 2 mL). The combined organic layers were washed with brine (30 X 4 mL), concentrated, and the residue purified by prep TLC eluting with MeOH in DCM (1/15) to afford ethyl 10'- formyl-9'-(oxetan-3-ylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (105 mg, 88% yield). LCMS (ESI) m/z: 424.4 (M + 1)+.
[00459] Step 4: Ethyl 10'-(difluoromethyl)-9'-(oxetan-3-ylmethoxy)-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000102_0002
[00460] To a solution of ethyl 10'-formyl-9'-(oxetan-3-ylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (105 mg, 0.25 mmol) in DCM (4 mL) was added DAST (161 mg, 1 .0 mmol) dropwise at 0 °C under N2. The mixture was stirred at rt overnight, diluted with DCM (30 mL), washed with sat.NaHC03 (20 mL), washed with brine (20 mL), and concentrated to afford ethyl 10'-(difluoromethyl)-9'- (oxetan-3-ylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (95 mg, 85% yield). LCMS (ESI) m/z: 446.7 (M + 1 )+. [00461] Step 5: 10'-(Difluoromethyl)-9'-(oxetan-3-ylmethoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000103_0001
[00462] To a solution of ethyl 10'-(difluoromethyl)-9'-(oxetan-3-ylmethoxy)-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate ( 95 mg, 0.21 mmol) in EtOH (6 mL) was added NaOH (34 mg, 0.85 mmol) in H20 (1 .5 mL). The mixture was stirred at rt for 2 h and concentrated. The mixture was acidified to pH= ~1 with 1 N HCI and extracted with DCM (10 mL X 3). The combined organic layers were washed with brine (30 mL), dried over Na2S04, concentrated, and the residue purified by reverse phase HPLC (C18, 30- 60 % acetonitrile in H20 with 0.1 % formic acid) to affordl O'- (difluoromethyl)-9'-(oxetan-3-ylmethoxy)-2'-oxo-2',7'-dihydrospiro [cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (30 mg, 34% yield) as a white powder. 1H NMR (400 MHz, DMSO-de) δ 8.82 (s, 1 H), 8.14 (s, 1 H), 7.35 (s, 2H), 7.07 (t, J = 54.7 Hz, 1 H), 4.86 - 4.64 (m, 2H), 4.47 (t, J = 6.0 Hz, 2H), 4.41 (d, J = 6.3 Hz, 2H), 3.64 - 3.37 (m, 3H), 2.80 - 2.56 (m, 2H), 2.16 - 1 .96 (m, 2H), 1 .98 - 1 .67 (m, 2H). LCMS (ESI) m/z: 418.3 (M +
[00463] Example 20
[00464] 10'-(Difluoromethyl)-9'-(3-methoxy-3-methylbutoxy)-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000103_0002
[00465] Step 1 : 3-Methoxy-3-methylbutyl 4-methylbenzenesulfonate
Figure imgf000103_0003
[00466] To a solution of 3-methoxy-3-methylbutan-1 -ol (1 .0 g, 8.47 mmol) in
DCM (20 mL) was added N,N-dimethylpyridin-4-amine (103 mg, 0.84 mmol), TEA (1 .7 g, 16.9 mmol) and 4-methylbenzenesulfonyl chloride (1 .61 g, 8.47 mmol). The mixture was stirred at rt overnight, concentrated, and the residue purified by column chromatography eluting with ethyl acatate in petroleum ether (1/4) to afford methoxy-3-methylbutyl 4- methylbenzenesulfonate (2.1 g, 91 % yield). LCMS (ESI) m/z: 273.4 (M + 1)+. [00467] Step 2: Ethyl 10'-formyl-9'-(3-methoxy-3-methylbutoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000104_0001
[00468] To a solution of ethyl 10'-formyl-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.28 mmol) in DMF (4 ml_) was added 3-methoxy-3-methylbutyl 4-methylbenzenesulfonate (153 mg, 0.56 mmol) and K2C03 (1 16 mg, 0.84 mmol). The mixture was heated at 90 °C overnight, cooled to room temperature, poured into water (30 ml_), and extracted with DCM (25 X 2 ml_). The combined organic layers were washed with brine (30 X 4 ml_), concentrated, and the residue purified by prep TLC eluting with MeOH in DCM (1 /15) to afford ethyl 10'- formyl-9'-(3-methoxy-3-methylbutoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (1 10 mg, 87% yield). LCMS (ESI) m/z: 454.5 (M + 1)+.
[00469] Step 3: Ethyl 10'-(difluoromethyl)-9'-(3-methoxy-3-methylbutoxy)-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000104_0002
[00470] To a solution of ethyl 10'-formyl-9'-(3-methoxy-3-methylbutoxy)-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (1 10 mg, 0.24 mmol) in DCM (4 mL) was added DAST (155 mg, 0.96 mmol) dropwise at 0 °C under N2. The mixture was stirred at rt overnight, diluted with DCM (30 mL), washed with sat.NaHC03 solution (20 mL), and washed with brine (20 mL). The organic phase was concentrated to afford ethyl 10'-(difluoromethyl)-9'-(3-methoxy-3-methylbutoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 87% yield). LCMS (ESI) m/z: 476.5 (M + 1)+. [00471] Step 4: 10'-(DifluoromethylV9'-(3-methoxy-3-methylbutoxyV2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000105_0001
[00472] To a solution of ethyl 10'-(difluoromethyl)-9'-(3-methoxy-3- methylbutoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate ( 85 mg, 0.18 mmol) in EtOH (6 mL) was added NaOH (29 mg, 0.72 mmol) in H20 (1 .5 mL). The mixture was stirred at rt for 2 h, concentrated, acidified to pH = ~1 with 1 N HCI, and extracted with DCM (10 mL X 3). The combined organic layers were washed with brine (30 mL), dried over Na2S04, concentrated, and the residue purified by reverse phase HPLC (C18, 30- 60 % acetonitrile in H20 with 0.1 % formic acid) to afford 10'- (difluoromethyl)-9'-(3-methoxy-3-methylbutoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (36 mg, 45% yield) as a white powder. 1H NMR (400 MHz, DMSO-de) δ 8.81 (s, 1 H), 8.12 (s, 1 H), 7.33 (s, 2H), 7.06 (t, J = 54.8 Hz, 1 H), 4.23 (t, J = 6.9 Hz, 2H), 3.43 (s, 2H), 3.13 (s, 3H), 2.82 - 2.58 (m, 2H), 2.12 - 1 .96 (m, 4H), 1 .95 - 1 .81 (m, 2H), 1 .20 (s, 6H). LCMS (ESI) m/z: 448.61 (M + 1)+.
[00473] Example 21
[00474] 10'-(Difluoromethyl)-9'-((3-methyloxetan-3-yl)methoxy)-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000105_0002
[00475] Step 1 : (3-Methyloxetan-3-yl)methyl 4-methylbenzenesulfonate
Figure imgf000105_0003
[00476] To a solution of (3-methyloxetan-3-yl)methanol (1 .02 g, 10 mmol) in
DCM (20 mL) was added N,N-dimethylpyridin-4-amine (122 mg, 1 mmol), TEA (2.0 g, 20 mmol) and 4-methylbenzenesulfonyl chloride (1 .9 g, 10 mmol). The mixture was stirred at rt for 3 h, concentrated, and the residue purified by column chromatography eluting with ethyl acatate in petroleum ether (1/4) to afford (3-methyloxetan-3-yl)methyl 4- methylbenzenesulfonate (1 .1 g, 43% yield). LCMS (ESI) m/z: 257.4 (M + 1)+. [00477] Step 2: Ethyl 10'-formyl-9'-((3-methyloxetan-3-yl)methoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000106_0001
[00478] To a solution of ethyl 10'-formyl-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.28 mmol) in DMF (4 ml_) was added (3-methyloxetan-3-yl)methyl 4-methylbenzenesulfonate (144 mg, 0.56 mmol) and K2C03 (1 16 mg, 0.84 mmol). The mixture was heated at 90 °C for 4 h, cooled to room temperature, poured into water (30 ml_), and extracted with DCM (25 X 2 ml_). The combined organic layers were washed with brine (30 X 4 ml_), concentrated, and the residue was purified by prep. TLC eluting with MeOH in DCM (1/15) to afford ethyl 10'- formyl-9'-((3-methyloxetan-3-yl)methoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 81 % yield). LCMS (ESI) m/z: 438.3 (M +
[00479] Step 3: Ethyl 10'-(difluoromethyl)-9'-((3-methyloxetan-3-yl)methoxy)-2'-oxo- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000106_0002
[00480] To a solution of ethyl 10'-formyl-9'-((3-methyloxetan-3-yl)methoxy)-
2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.23 mmol) in DCM (4 mL) was added DAST (185 mg, 1 .15 mmol) dropwise at 0 °C under N2. The mixture was stirred at rt overnight, diluted with DCM (30 mL), washed with sat.NaHC03 solution (20 mL), washed with brine (20 mL), and concentrated to afford ethyl 10'-(difluoromethyl)-9'-((3-methyloxetan-3-yl)methoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (90 mg, 86% yield). LCMS (ESI) m/z: 460.2 (M + 1)+.
[00481] Step 4: 10'-(Difluoromethvn-9'-((3-methyloxetan-3-vnmethoxyV2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000107_0001
[00482] To a solution of ethyl 10'-(difluoromethyl)-9'-((3-methyloxetan-3- yl)methoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate ( 90 mg, 0.19 mmol) in EtOH (6 mL) was added NaOH (31 mg, 0.78 mmol) in H20 (1 .5 mL). The mixture was stirred at rt for 2 h, concentrated, acidified to pH = ~1 with 1 N HCI, and extracted with DCM (10 mL X 3). The combined organic layers were washed with brine (30 mL), dried over Na2S04, concentrated, and the residue purified by reverse phase HPLC (C18, 30- 55 % acetonitrile in H20 with 0.1 % formic acid) to afford 10'- (difluoromethyl)-9'-((3-methyloxetan-3-yl)methoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (38 mg, 45% yield) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1 H), 8.15 (s, 1 H), 7.35 (d, J = 4.2 Hz, 2H), 7.08 (t, J = 54.7 Hz, 1 H), 4.52 (d, J = 5.8 Hz, 2H), 4.34 (d, J = 5.9 Hz, 2H), 4.26 (s, 2H), 3.43 (s, 2H), 3.32 (s, 3H), 2.82 - 2.57 (m, 2H), 2.17 -1 .96 (m, 2H), 1 .99 -1 .68 (m, 2H). LCMS (ESI) m/z: 432.5 (M + 1)+.
[00483] Example 22
[00484] 3-Hvdroxy-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000107_0002
[00485] Step 1 : 4-Methoxy-3-(3-methoxypropoxy)benzaldehvde
Figure imgf000107_0003
[00486] To a stirred solution of 3-hydroxy-4-methoxybenzaldehyde (40 g,
0.17 mol) in MeCN (500 mL ) were added 1 -bromo-3-methoxypropane (32 g, 0.21 mol) and K2C03 (70 g, 0.51 mol). The mixture was refluxed overnight, cooled to room temperature and quenched with H20, and extracted with EtOAc(100 mL x 3). The combined organic layers were washed with brine, dried over Na2S04, and concentrated to give crude 4- methoxy-3-(3-methoxypropoxy)benzaldehyde (50 g, 84% yield) as a white solid. LCMS (ESI) m/z: 225.3 (M + 1)+. [00487] Step 2: (4-Methoxy-3-(3-methoxypropoxy)phenyr)methanol
Figure imgf000108_0001
[00488] To a 0 °C solution of 4-methoxy-3-(3-methoxypropoxy)benzaldehyde
(50 g, 0.22 mol) in MeOH (500 mL ) was added NaBH4 (16 g, 0.44 mol) in portions. The mixture was stirred at 0 °C for 2h, concentrated, and portioned between EtOAc and H2O. The organic layer was separated and washed with brine, dried over Na2S04, and concentrated to give crude (4-methoxy-3-(3-methoxypropoxy)phenyl)methanol (45 g, 89% yield) as a white solid. LCMS (ESI) m/z: 227.1 (M + 1)+.
[00489] Step 3: -(Bromomethyl)-1 -methoxy-2-(3-methoxypropoxy)benzene
Figure imgf000108_0002
[00490] Bromotrimethylsilane (17.5 mL, 0.13 mol) was added dropwise to a 0
°C solution of (4-methoxy-3-(3-methoxypropoxy)phenyl)methanol (20 g, 0.088 mol) in CHC (200 mL). The mixture was stirred at rt for 30 minutes and then stood for 1 h. The upper liquid was collected and concentrated. The residue was purified by flash chromatography (silica gel, 0-15% EtOAc in petroleum ether) to afford 4-(bromomethyl)-1 -methoxy-2-(3- methoxypropoxy)benzene (22 g, 86.3% yield) as a white solid. 1H NMR (400 MHz, CDCI3) δ 6.87 (m, 2H), 6.75 (d, J = 8.0 Hz, 1 H), 4.42 (s, 2H), 4.06 (t, J = 6.5 Hz, 2H), 3.79 (s, 3H), 4.06 (t, J = 6.1 Hz, 2H), 3.29 (s, 3H), 4.06 (p, J = 6.3 Hz, 2H).
[00491] Step 4: Methyl 3-(benzyloxy)-1 -(4-methoxy-3-(3- methoxypropoxy)benzyl)cvclobutane-1 -carboxylate
Figure imgf000108_0003
[00492] Methyl 3-(benzyloxy)cyclobutane-1 -carboxylate (23.6 g, 107 mmol) in THF (80 mL) was added dropwise to a solution of lithium diisopropylamide (2.0 M in THF, 68 mL, 136 mmol) in THF (270 mL) at -78°C. The mixture was stirred at this temperature for 1 h. 4- (Bromomethyl)-1 -methoxy-2-(3-methoxypropoxy) benzene (26.2 g, 90.6 mmol ) in THF (100 mL) was added dropwise and the mixture was stirred at -78°C for 1 h. The cold bath was removed and the mixture was stirred at room temperature overnight. The mixture was cooled to 0 °C, quenched with water, extracted with ethyl acetate, dried over sodium sulfate, and concentrated under vacuum. The residue was purified via silica gel chromatography (5- 30% EtOAc/PE) to give methyl 3-(benzyloxy)-1 -(4-methoxy-3-(3- methoxypropoxy)benzyl)cyclobutane-1 -carboxylate (33 g, 85% yield ) as colorless oil. LCMS (ESI) m/z: 429.27 (M + 1)+.
[00493] Step 5: 3-(Benzyloxy)-1 -(4-methoxy-3-(3- methoxypropoxy)benzvDcvclobutane
[00494] carboxylic acid
Figure imgf000109_0001
[00495] To a solution of methyl 3-(benzyloxy)-1 -(4-methoxy-3-(3- methoxypropoxy)benzyl) cyclobutane -1 -carboxylate (33 g, 77 mmol) in THF (200 ml), methanol (100 ml), and water (50 ml)was added LiOH (20 g, 476 mmol), The mixture was stirred at 65°C for 3 h, cooled to room temperature, concentrated, and extracted with Et20. The aqueous layer was acidified with 1 N HCI to pH = ~3. Water and EtOAc were added and the organic layer was separated. The organic phase was concentrated to give 3- (benzyloxy)-1 -(4-methoxy-3-(3- methoxypropoxy)benzyl)cyclobutanecarboxylic acid (19 g, 59.6% yield) as a yellow solid. LCMS (ESI) m/z: 415.2 (M + 1)+.
[00496] Step 6: 4-((3-(Benzyloxy)-1 -isocvanatocvclobutyl)methyl)-1 -methoxy-2-(3- methoxypropoxy)benzene
Figure imgf000109_0002
[00497] A solution of 3-(benzyloxy)-1 -(4-methoxy-3-(3- methoxypropoxy)benzyl)cyclobutanecarboxylic acid (18.3 g, 44.1 mmol), DPPA (15.8 g, 57.4 mmol), Et3N (8 mL, 57.4mmol) in toluene (200 mL) was stirred at 130 °C under N2 for 3 h. The mixture was concentrated to give crude 4-((3-(benzyloxy)-1 - isocyanatocyclobutyl)methyl)-1 -methoxy-2-(3-methoxypropoxy)benzene, which was used in the next step without purification. LCMS (ESI) m/z: 412.2 (M + 1 )+.
[00498] Step 7: 3-(Benzyloxy)-1 -(4-methoxy-3-(3-methoxypropoxy) benzvDcvclo butanamine
Figure imgf000109_0003
[00499] Crude 4-((3-(benzyloxy)-1 -isocyanatocyclobutyl)methyl)-1 -methoxy-
2-(3-methoxypropoxy)benzenewas dissolved in THF(150 mL). Concentrated HCI (40 mL) was added slowly and the mixture heated at 60 °C for 3 h (Caution: this reaction released bubbles). The mixture was cooled to room temperature, basified with aqueous NaHC03 to pH = 8, and extracted with EtOAc (150 mL x 2). The organic layer was separated, dried over Na2S04, and concentrated to afford crude 3-(benzyloxy)-1 -(4-methoxy-3-(3- methoxypropoxy)benzyl)cyclobutanamine, which was used in the next step without purification. LCMS (ESI) m/z: 386.3 (M + 1)+.
[00500] Step 8: N-(3-(Benzyloxy)-1 -(4-methoxy-3-(3- methoxypropoxy)benzyl)cvclobutyl) formamide
Figure imgf000110_0001
[00501] Acetic formic anhydride (19.4 g, 221 mmol) was added dropwise to a solution of 3-(benzyloxy)-1 -(4-methoxy-3-(3-methoxypropoxy)benzyl)cyclobutanamine
[00502] (18.2 g, 44.1 mmol) and TEA (8.9 g, 88.2 mmol) in THF(200 mL) at 0°C. The mixture was stirred at 0°C for 1 h and then partitioned between ethyl acatate and water. The organic layer was separated and the aqueous layer extracted with EA. The combined organic layers were dried over Na2S04 and concentrated. The residue was purified by flash chromatography (silica gel, 0-40 % EtOAc in petroleum ether) to afford N-(3-(benzyloxy)-1 - (4-methoxy-3-(3-methoxypropoxy)benzyl)cyclobutyl) formamide (14.1 g, 77 % yield over three steps). LCMS (ESI) m/z: 414.3 (M + 1)+.
[00503] Step 9: 3-(Benzyloxy)-7'-methoxy-6'-(3-methoxypropoxy)-4'H-
Figure imgf000110_0002
[00504] To a 0°C solution of N-(3-(benzyloxy)-1 -(4-methoxy-3-(3- methoxypropoxy)benzyl) cyclobutyl)formamide (14.1 g, 34.1 mmol) in acetonitrile (120 mL) was added POCI3 (5.22 g, 34.1 mmol) dropwise. The mixture was heated to 90°C for 1 h and cooled to room temperature. The mixture was concentrated and the residue dissolved in DCM. The organic mixture was washed with water, dried over Na2S04, and concentrated to give crude 3-(benzyloxy)-7'-methoxy-6'-(3-methoxypropoxy)-4'H-spiro[cyclobutane-1 ,3'- isoquinoline]. LCMS (ESI) m/z: 396.2 (M + 1 )+.
[00505] Step 10: Ethyl 3-(benzyloxy)-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo- 1 ',2',7',1 1 b'-tetrahvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'- carboxylate
Figure imgf000111_0001
[00506] To a solution of 3-(benzyloxy)-7'-methoxy-6'-(3-methoxypropoxy)-
4'H-spiro[cyclobutane-1 ,3'-isoquinoline] (13g, 136 mmol) in ethanol (150 mL) was added ethyl 2-(ethoxymethylene)-3-oxobutanoate (25.3 g, 136 mmol). The resulting mixture was heated to 90°C overnight. After removing the solvent, the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to afford ethyl 3- (benzyloxy)-I O'- methoxy-9'-(3-methoxypropoxy)-2'-oxo-1 ',2',7',1 1 b'-tetrahydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'- carboxylate (12 g, 65.6% yield over two steps). LCMS (ESI) m/z: 536.3 (M + 1)+.
[00507] Step 1 1 : Ethyl 3-(benzyloxy)-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo- 2',7'-dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000111_0002
[00508] To a stirred solution of ethyl 3-(benzyloxy)-10'-methoxy-9'-(3- methoxypropoxy)-2'-oxo-1 ',2',7',1 1 b'-tetrahydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'- carboxylate (12 g, 22mmol) in DME (120 mL) was added p-chloranil (4.7 g, 19 mmol). The mixture was heated at 90°C overnight and cooled to room temperature. The mixture was concentrated and the residue purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to give ethyl 3-(benzyloxy)-10'-methoxy-9'-(3-methoxypropoxy)-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (7.35 g, 61 .7% yield). LCMS (ESI) m/z: 534.4 (M + 1)+.
[00509] Step12: Ethyl 3-hvdroxy-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000111_0003
[00510] To a solution of ethyl 3-(benzyloxy)-10'-methoxy-9'-(3- methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoqu'
no carboxylate (7 g, 13 mmol) in EtOH (120 mL) was added palladium (700 mg, 5% on carbon (wetted with ca. 55% Water)). The mixture was stirred at 80°C under H2 (15 psi) overnight. The mixture was filtered and the filtrate concentrated to afford ethyl 3-hydroxy-10'-methoxy- 9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate (3.8 g, 65.5% yield). LCMS (ESI) m/zd: 444.2(M + 1 )+.
[00511] Step 13: 3-Hvdroxy-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000112_0001
[00512] To a solution of ethyl 3-hydroxy-10'-methoxy-9'-(3-methoxypropoxy)-
2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (160 mg, 0.36 mmol) in THF/EtOH/H20 (2 mL /1 mL /0.5 mL) was added LiOH (61 mg, 1 .44 mmol). The mixture was stirred at r.t for 1 hr, acidified with 1 N HCI, and extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered, concentrated and the residue purified by reverse phase HPLC (C18, 0-100 % MeCN in H20 with 0.1 % formic acid) to afford 3-hydroxy-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (30 mg, 20% yield) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 10.15 (br, 1 H), 8.78 (s, 1 H), 7.53- 7.37 (m, 2H), 7.03 (s, 1 H), 5.44 (d, J = 16.5 Hz, 1 H),4.39-4.28(m, 1 H),4.16-4.04 (m, 3H), 3.87 (s, 3H),3.48 (t, J = 6.2 Hz, 2H), 3.25 (s, 3H), 3.13 (s, 1 H),2.86-2.77 (m,1 H), 2.48-2.40 (m, 2H), 2.1 1 (dd, J = 13.7,4.8 Hz, 1 H), 2.02-1 .97 (m, 2H). LCMS (ESI) m/z: 416.4 (M + 1 )+.
[00513] Example 23
[00514] 3-Fluoro-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid (isomer 1)
Figure imgf000112_0002
[00515] Step 1 : Ethyl 3-fluoro-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000113_0001
[00516] To a solution of ethyl 3-hydroxy-10'-methoxy-9'-(3- methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (80 mg, 0.18 mmol) in DCM (1 mL) was added DAST (212.4 mg, 0.5 mmol). The mixture was stirred at r.t for 3 h, quenched with ice-water, and extracted with DCM . The organic layer was washed with brine, dried over Na2S04, concentrated, and the residue purified by flash chromatography (silica gel, 0 - 50 % DCM in MeOH) to afford ethyl 3-fluoro-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylate (50 mg, 62 % yield). LCMS (ESI) m/z: 446.3 (M +
[00517] Step 2: 3-Fluoro-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid (isomer 1)
Figure imgf000113_0002
isomer 1
[00518] To a solution of ethyl 3-fluoro-10'-methoxy-9'-(3-methoxypropoxy)-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (50 mg, 0.1 mmol) in THF (2 mL) was added LiOH (1 1 mg, 0.45 mmol) and H20 (0.5 ml). The mixture was stirred at room temperature for 2 h and acidified to pH = 7, concentrated, and the residue was purified by reverse phase HPLC (C18, 30-100% MeCN in H20 with 0.1 % formic acid). Fractions for corresponding to one peak were concentrated to afford 3-fluoro-10'- methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid (isomer 1) (5.4 mg, 21 % yield) as a white powder. 1H NMR (400 MHz, DMSO-de) δ 8.76 (s, 1 H), 7.50 (s, 1 H), 7.43 (s, 1 H), 6.98 (s, 1 H), 5.27-5.07 (m,1 H) , 4.10 (t,J=6.4 Hz, 2H), 3.88 (s, 3H), 3.48(t, J=6.2 Hz, 2H), 3.25 (s, 3H), 3.14(s, 2H), 2.89-2.81 (m, 2H), 2.67-2.62 (m, 2H), 2.02-1 .97 (m, 2H). LCMS (ESI) m/z:418.3 (M + 1)+. [00519] Example 24
[00520] 3-Fluoro-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid (isomer 2)
Figure imgf000114_0001
isomer 2
[00521] Step 1 : 3-Fluoro-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid (isomer 2)
[00522] Fractions corresponding to a second peak in the reverse phase
HPLC purification described above were concentrated to give 3-fluoro-10'-methoxy-9'-(3- methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylic acid (isomer 2) (3.1 mg, 21 % yield) as a white powder. 1H NMR (400 MHz, DMSO-de) δ 8.74 (s, 1 H), 7.49 (s, 1 H), 7.45 (s, 1 H), 7.09 (s, 1 H), 5.41 -5.39 (m,1 H), 5.27- 5.25 (m, 1 H), 4.12 (t, J=6.5 Hz, 2H), 3.88 (s, 3H), 3.48(t, J=6.2 Hz, 2H), 3.36 (s, 2H), 3.25(s, 3H), 3.12-3.06 (m, 2H), 2.44-2.38 (m, 2H), 2.01 -1 .97 (m, 2H). LCMS (ESI) m/z: 418.3 (M + 1)+.
[00523] Example 25
[00524] 10'-Methoxy-9'-(3-methoxypropoxy)-2'.3-dioxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000114_0002
[00525] Step 1 : 3-Hvdroxy-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000114_0003
[00526] To a solution of ethyl 3-hydroxy-10'-methoxy-9'-(3-methoxypropoxy)-
2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg,
0.23 mmol) in THF/EtOH/H20(2 mL /1 mL /0.5 mL) was added LiOH (17 mg, 0.69 mmol). The mixture was stirred at r.t for 1 hr, acidified with 1 N HCI, and extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, and concentrated to give crude 3-hydroxy-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (80 mg, 80% yield ) as a white powder. LCMS (ESI) m/z: 416.4 (M + 1)+.
[00527] Step 2: 10'-Methoxy-9'-(3-methoxypropoxy)-2'.3-dioxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000115_0001
[00528] To a solution of 3-hydroxy-10'-methoxy-9'-(3-methoxypropoxy)-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (80 mg, 0.19 mmol) in DCM (1 mL) at 0 °C was added Dess-Martin reagent (213mg, 0.5 mmol). The mixture was stirred at 0 °C for 3 h and filtered. The filtrate was concentrated and the residue purified by reverse phase HPLC (C18, 30-100% MeCN in H20 with 0.1 % formic acid) to afford 10'-methoxy-9'-(3-methoxypropoxy)-2',3-dioxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (13.3 mg, 17% yield) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 8.90 (s, 1 H), 7.52 (s, 1 H), 7.49 (s, 1 H), 7.05 (s, 1 H), 4.10 (t,J=6.4 Hz, 2H), 3.96 (d, J=19.0 Hz, 2H), 3.89 (s, 3H), 3.47(t, J=6.1 Hz, 2H), 3.37 (s, 2H), 3.25(s, 3H), 3.15 (d, J=19.2 Hz, 2H), 2.02-1 .96 (m, 2H). LCMS (ESI) m/z: 414.3 (M + 1)+.
[00529] Example 26
[00530] 3.10'-Dimethoxy-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000115_0002
[00531] Step 1 : 3,10'-Dimethoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000116_0001
[00532] To a solution of ethyl 3-hydroxy-10'-methoxy-9'-(3-methoxypropoxy)-
2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (1 1 1 mg, 0.25 mmol) in DMF (1 .0 mL) was added NaH (60% mixture with mineral oil, 25 mg, 0.625 mmol). The mixture was stirred at rt for 30 min before Mel (19 mg, 0.3 mmol) was added. The mixture was stirred at rt for 3 h and diluted with MeOH/H20 (0.5 mL / 0.5 mL). The mixture continued to stir for 2 h was then acidified with 1 N HCI. The mixture was extracted with CH2CI2 (3x5 mL). The combined organic layers were washed with brine, dried over Na2S04, concentrated and the residue purified by reverse phase HPLC (C18, 0-90% MeCN in H20 with 0.1 % formic acid) to afford 3,10'-dimethoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (38.5 mg, 35% yield) as an off-white powder. 1H NMR (400 MHz, CDCI3) δ 16.1 1 - 15.85 (s, 1 H), 8.93 - 8.82 (s, 1 H), 7.18 - 7.12 (s, 1 H), 7.09 - 7.02 (s, 1 H), 6.83 - 6.76 (s, 1 H), 4.25 - 4.17 (m, 2H), 4.16 - 4.1 1 (m, 0.5H), 3.99 - 3.91 (s, 3H), 3.91 - 3.86 (m, 0.5H), 3.59 (t, J = 5.6 Hz, 2H), 3.37 (s, 3H), 3.32 - 3.23 (m, 4H), 3.05 (s, 1 H), 2.88 - 2.77 (m, 1 H), 2.63 - 2.47 (m, 2H), 2.30 (dd, J = 14.4, 4.0 Hz, 1 H), 2.15 (p, J = 6.0 Hz, 2H). LCMS (ESI) m/z: 430.3 (M +
[00533] Example 27
[00534] 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'-dihvdrospiro[cyclohexane-1 ,6'- pyrido[2,1 -alisoquinolin -3'-carboxylic acid
Figure imgf000116_0002
[00535] Step 1 : Benzyl 3-(benzyloxy)-4-bromobenzoate
Figure imgf000116_0003
[00536] To a solution of 4-bromo-3-hydroxybenzoic acid (90 g, 418.6 mmol) and K2C03 (173 g, 1255.8 mmol) in MeCN (1800 mL) was added BnBr (142 g, 837mmol) dropwise at room temperature over 30 min. The mixture was stirred at 65 °C overnight. The mixture was cooled to room temperature and water (900 mL) added. The mixture was extracted 2 times with ethyl acatate (1800 mL). The combined organic layers were washed with bine (900 mL), dried over MgS04 (500 g), concentrated, and the residue crystallized with MeOH (1800 mL) for 2 h at 0 °C. The precipitate collected by filtration and washed with cold MeOH (100 mL) to afford benzyl 3-(benzyloxy)-4-bromobenzoate (140 g, 85%). LCMS (ESI) m/z: 397.1 / 399.2 (M/M+2)+.
[00537] Step 2: -(Benzyloxy)-4-bromophenyl) methanol
Figure imgf000117_0001
[00538] To a suspension of LiAlhU (13.8 g, 3646 mmol) in THF (130 mL) at -20 °C was added a solution of (3-(benzyloxy)-4-bromophenyl)methanol (144 g, 364 mmol) in THF (1500 mL) dropwise over 2 h. The mixture was stirred at -20 °C for 2 h. Water (13.8 g) was added dropwise over 30 min at 0 °C. 40% Aqueous sodium hydroxide (13.8 g) was added dropwise over 30 min and stirred for another 30 min. The precipitate was filtered away and the filtrate concentrated. The residue was dissolved in ethyl acatate (1500 mL), washed with brine, dried over MgS04, and concentrated. The residue was crystallized with petroleum ether (1500 mL) for 2 h at 0 °C. The white solid was collected by filtration and washed with petroleum ether (500 mL) to afford (3-(benzyloxy)-4-bromophenyl)methanol (72 g, 68% yield). Ή NMR (400 MHz, DMSO-d6) δ 7.51 (dd, J = 12.9, 7.6 Hz, 3H), 7.42 (dd, J = 10.1 , 4.7 Hz, 2H), 7.36 - 7.32 (m, 1 H), 7.17 (d, J = 1 .4 Hz, 1 H), 6.90 - 6.80 (m, 1 H), 5.29 (t, J = 5.7 Hz, 1 H), 5.19 (s, 2H), 4.47 (t, J = 5.7 Hz, 2H).
[00539] Step 3: -(Benzyloxy)-4-bromobenzyl 4-methylbenzenesulfonate
Figure imgf000117_0002
[00540] To a solution of (3-(benzyloxy)-4-bromophenyl)methanol (62 g, 213 mmol) in DCM (620 mL) was added DMAP (1 .3 g, 10.6 mmol) and DIPEA (82.2 g, 637mmol). A solution of TsCI (49 g, 254.8 mmol) in DCM (500 mL) was added dropwise at 0 °C and stirred at room temperature for 2 h. The mixture was concentrated and the residue purified by flash chromatography (silica gel, 0-50% EtOAc in petroleum ether) to afford 3- (benzyloxy)-4-bromobenzyl 4-methylbenzenesulfonate (90 g, 95% yield). LCMS (ESI) m/z: 447.I / 449.4 (M/M+2)+. [00541] Step 4: Methyl 1 -(3-(benzyloxy)-4-bromobenzvDcvclohexane-1 -carboxylate
Figure imgf000118_0001
[00542] To a stirred solution of methyl cyclohexanecarboxylate (34.4 g,
241 .4 mmol) in anhydrous THF (350 mL) at -78 °C, was added LDA (181 mL, 2 M in THF/hexane) dropwise. The mixture was stirred for 1 hour before a solution of 3- (benzyloxy)-4-bromobenzyl 4-methylbenzenesulfonate (90 g, 201 .2 mmol) in anhydrous THF (1 L) was added dropwise. The mixture was stirred at -78 °C for 30 minutes and warmed to room temperature gradually. The mixture was partitioned between EtOAc and saturated NH4CI and extracted with EtOAc. The organic phase was dried over Na2S04, concentrated, and the residue purified by flash chromatography (silica gel, 0-10% EtOAc in petroleum ether) to afford methyl 1 -(3-(benzyloxy)-4-bromobenzyl)cyclohexane-1 - carboxylate (76 g, 87.6% yield). ). Ή NMR (400 MHz, CDCI3) δ 7.48 - 7.43 (m, 2H), 7.42 - 7.34 (m, 3H), 7.33 - 7.27 (m, 1 H), 6.61 (d, J = 1 .8 Hz, 1 H), 6.52 (dd, J = 8.0, 1 .8 Hz, 1 H), 5.12 (s, 2H), 3.56 (s, 3H), 2.70 (s, 2H), 2.02 - 1 .94 (m, 2H), 1 .59 - 1 .50 (m, 3H), 1 .30 - 1 .15 (m, 5H).
[00543] Step 5: -(3-(benzyloxy)-4-bromobenzyl)cyclohexane-1 -carboxylic acid
Figure imgf000118_0002
[00544] To a solution of methyl 1 -(3-(benzyloxy)-4- bromobenzyl)cyclohexane-1 -carboxylate (76 g, 176.2 mmol) in DMSO (800 mL) was added KOH (39.6 g, 705mmol). The mixture was heated at 120 °C for 5 hours. The reaction was acidified with 1 N HCI to pH 6 at ice-water bath, extracted with EtOAc, the organic layers were dried over Na2S04, filtered and concentrated, the residue was purified by flash chromatography (silica gel, 0-40% EtOAc in petroleum ether) to afford 1 -(3-(benzyloxy)-4- bromobenzyl)cyclohexane-1 -carboxylic acid (40.3 g, 56.7% yield). LCMS (ESI) m/z 401 .32, 403.3 (M-2, M)-
[00545] Step 6:
Figure imgf000118_0003
[00546] At 0 °C, to a solution of 1 -(3-(benzyloxy)-4-bromobenzyl)cyclohexane-1 - carboxylic acid (40.3 g, 99.9 mmol), TEA (13.1 g, 130 mmol) in toluene (500 mL) was added DPPA (37.78 g, 130 mmol). The resulting mixture was stirred at 120 °C for 5 hours. The reaction mixture was partitioned between toluene and water. The organic layer was separated, the aqueous layer was extracted with toluene, and the combined organic layers were washed with brine, filtered and concentrated to 100 ml_ volume. To the solution of 2- (benzyloxy)-1 -bromo-4-((1 -isocyanatocyclohexyl) methyl)benzene in toluene at 0 °C, was added conc.HCI (40 ml). The mixture was stirred at room temperature for 2 hours and the solvent was removed. The residue was dissolved in DCM and treated with Sat. NaHC03. The organic layer was separated and the aqueous layer extracted with DCM. The combined organic layers were dried over Na2S04, concentrated, and the residue was purified by flash chromatography (silica gel, 0-100% EtOAc in petroleum ether) to afford 1 -(3-(benzyloxy)-4- bromobenzyl)cyclohexan-1 -amine (16 g, 42% yield). LCMS (ESI) m/z: 374.2, 376.2
(M,M+2)+.
[00547] Step 7: -(1 -(3-(Benzyloxy)-4-bromobenzyl)cvclohexyl)formamide
Figure imgf000119_0001
[00548] To a solution of 1 -(3-(benzyloxy)-4-bromobenzyl)cyclohexan-1 - amine (16 g, 42 mmol) in anhydrous THF (200 ml_) at 0 °C, was added acetic formic anhydride (36.9 g, 420 mmol) dropwise. The resulting mixture was stirred at the same temperature for 30 minutes. The mixture was partitioned between DCM and water and the organic layer was separated. The aqueous layer was extracted with DCM. The combined organic layers were dried over Na2S04, concentrated, and the residue purified by flash chromatography (silica gel, 0-50% EtOAc in petroleum ether) to afford /V-(1 -(3-(benzyloxy) 4-bromobenzyl)cyclohexyl)formamide (13 g, 76.9% yield). LCMS (ESI) m/z: 402.2 ,404.2 (M, M+2)+.
[00549] Step 8: 6'-(Benzyloxy)-7'-bromo-4'H-spiro[cyclohexane-1 ,3'-isoquinolinel
Figure imgf000119_0002
[00550] To a solution of Λ/-(1 -(3-(benzyloxy)-4- bromobenzyl)cyclohexyl)formamide (13 g, 32.3 mmol) in acetonitrile (150 ml_) at 0 °C was added POCI3 (5.9 g, 38.77 mmol) dropwise. The mixture was stirred at r. t. for 8 hours and concentrated. The residue was dissolved in DCM and washed with water. The organic layer was dried over Na2S04, concentrated, and the residue purified by flash chromatography (silica gel, 0-80% EtOAc in petroleum ether) to afford 6'-(benzyloxy)-7'-bromo-4'H- spiro[cyclohexane-1 ,3'-isoquinoline] (1 1 g, 88.6% yield). LCMS (ESI) m/z: 384.2 ,386.2 (M, M+2)+.
[00551] Step 9: Ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-1 '.2'.7'.1 1 b1- tetrahvdrospiro[cvclohexane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000120_0001
[00552] To a solution of 6'-(benzyloxy)-7'-bromo-4'H-spiro[cyclohexane-1 ,3'- isoquinoline] (1 1 g, 28.6 mmol) in EtOH (120 ml_) was added ethyl 2-(ethoxymethylene)-3- oxobutanoate (31 .9 g, 172mmol). The mixture was heated at 84 °C for 12 hours. The mixture was cooled, concentrated, and the residue purified by flash chromatography (silica gel, 0-100% EtOAc in petroleum ether) to afford ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo- 1 ',2',7',1 1 b'-tetrahydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (7.13 g, 47.5% yield). LCMS (ESI) m/z: 524.2 ,526.1 (M, M+2)+.
[00553] Step 10: Ethyl 9'-(benzyloxy)-10'-bromo^'-oxo^'J1- dihydrospirofcyclohexane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000120_0002
[00554] To a stirred solution of ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-
1 ',2',7',1 1 b'-tetrahydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (7.13 g, 13.6 mmol) in DME (100 mL) was added p-chloroanil (2.84 g,1 1 .5 mmol). The mixture was heated at 80 °C for 5 hours. The mixture was cooled to room temperature, concentrated, and the residue purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to afford ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'-dihydrospiro[cyclohexane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylate (6.8 g, 95.7% yield). LCMS (ESI) m/z: 522.2 ,524.2 (M, M+2)+. [00555] Step 1 1 : 9'-(Benzyloxy)-10'-bromo-2'-oxo-2\7' lihvdrospiro[cvclohexane-
Figure imgf000121_0001
[00556] To a solution of ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate(100 mg, 0.19 mmol) in THF (5 ml) was added NaOH (31 mg, 0.76 mmol) in H20 (1 ml). The mixture was stirred at room temperature for 2 h and acidified to pH = ~5 with 1 N HCI. The mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50ml_ x 2). The combined organic layers were dried over anhydrous Na2S04, concentrated, and the residue purified by Gilson (C18 column, 30%1 100% MeCN in H20, with 0.1 % formic acid) to give 9'- (benzyloxy)-10'-bromo-2'-oxo-2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylic acid (46.4 mg, 49% yield) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1 H), 8.33 (s, 1 H), 7.53 - 7.34 (m, 7H), 5.31 (s, 2H), 3.30 (s, 2H), 2.01 - 1 .92 (m, 2H), 1 .80 - 1 .73 (m, 2H), 1 .72 - 1 .64 (m, 3H), 1 .62 - 1 .52 (m, 2H), 1 .38 - 1 .30 (m, 1 H). LCMS (ESI) m/z: 494.3 ,496.4 (M, M+2)+.
[00557] Example 28
[00558] 10'-Methyl-2'-oxo-9'-(piperidin-1 -yl)-2',7'-dihvdrospiro[cvclohexane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000121_0002
[00560] To a solution of ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (500 mg, 0.96 in toluene (4 mL) was added methyboronic acid (172 mg, 0.2.87 mmol), Pd(PPh3)4 (50 mg), and Cs2C03 (1 .6 g, 4.8 mmol) in H20 (1 mL). After stirring at 85 °C for 6 h, the reaction mixture was cooled down to room temperature, the solvents were removed under vacuum to give the crude product which was purified by flash chromatography (silica gel, 0-50% DCM in MeOH) to afford the title compound (440 mg, 100 % yield). LCMS (ESI) m/z: 458.42 (M + 1 )+.
[00561] Step 2: Ethyl 9'-hvdroxy-10'-methyl-2'-oxo-2',7'-dihvdrospiro[cyclohexane-
Figure imgf000122_0001
[00562] A solution of ethyl 9'-(benzyloxy)-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (440 mg, 0.96 mmol) in TFA (4 mL) was stirred at stirred at 60 °C for 3 h. The reaction mixture was cooled down to room temperature and the solvents were removed under vacuum to give the product (320 mg, 91 % yield). LCMS (ESI) m/z: 368.32 (M + 1)+.
[00563] Step 3: Ethyl 10'-methyl-2'-oxo-9'-(((trifluoromethyl)sulfonyl)oxy)-2'.7'- dihydrospirofcyclohexa -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000122_0002
[00564] To a solution of ethyl 9'-hydroxy-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (160 mg, 0.435 mmol) in DCM (10 mL) was added N,N-dimethylpyridin-4-amine (5 mg, 0.044 mmol), TEA (133 mg, 1 .32 mmol) and 1 ,1 ,1 -trifluoro-N-phenyl-N-
((trifluoromethyl)sulfonyl)methanesulfonamide (233 mg, 0.65mmol) under N2 atmosphere. The reaction mixture was stirred at room temperature for 4 h. The solvents were concentrated and the residue was purified by column chromatography to afford the title compound (200 mg, 92% yield). LCMS (ESI) m/z: 500.27 (M + 1 )+.
[00565] Step 4: Ethyl 1 O'-methyl^'-oxo-g'-foiperidin-l -νη-2'.7'- dihydrospirofcyclohexane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000123_0001
[00566] A mixture of ethyl 10'-methyl-2'-oxo-9'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate
[00567] (150 mg, 0.3 mmol), Pd2(dba)3 (27mg,0.03 mmol), BINAP (28 mg, 0.045 mmol),Cs2C03 (591 mg,1 .8 mmol) in toluene (5 ml), piperidine (51 mg, 0.6 mmol) was stirred at 100 °C for 2 hours. The mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50ml_x2). The combined organic layers were dried over anhydrous Na2SC>4 and concentrated. The residue was purified by column chromatography (silica gel,0~10%MeOH in DCM) to afford the title compound. (20 mg, 15% yield). LCMS (ESI) m/z: 435.37 (M + 1 )+.
[00568] Step 5: 10'-Methyl-2'-oxo-9'-(piperidin-1 -ylV2',7'-dihvdrospiro[cvclohexane-
Figure imgf000123_0002
[00569] To a solution of ethyl 10'-methyl-2'-oxo-9'-(piperidin-1 -yl)-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (20 mg, 0.048 mmol) in MeOH (3 ml) was added LiOH (4 mg, 0.14mmol) in H20 (0.5ml). After stirring at room temperature for 20 minutes, the reaction mixture was quenched with 1 N HCI to pH 6, then was extracted with 15% IPA in DCM twice. The combined organic layer was dried over anhydrous Na2S04, filtered and concentrated. The residue was purified by reverse phase reverse phase HPLC (Gilson, C18, 20%~100% MeCN in H20,with 0.1 % formic acid) to afford the title compound. (6 mg, 31 % yield) as a yellow solid. LCMS (ESI) m/z: 407.3 (M + 1)+. Ή NMR (400 MHz, CDCI3) δ 8.91 (s, 1 H), 7.53 (s, 1 H), 7.10 (s, 1 H), 6.79 (s, 1 H), 3.12 (s, 2H), 2.97 - 2.92 (m, 4H), 2.34 (s, 3H), 1 .90 - 1 .85 (m, 4H), 1 .85 - 1 .72 (m, 8H), 1 .64 - 1 .62 (m, 2H), 1 .31 - 1 .29 (m, 2H).
[00570] Example 29 [00571] 9'-(CvclopropylmethoxyV10'-methyl-2'-oxo-2',7'-dihvdrospiro[cyclohexane- 1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000124_0001
[00572] Step 1 : Ethyl 9'-(cvclopropylmethoxyV10'-methyl-2'-oxo-2'.7'- dihvdrospiro[cvclohexane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000124_0002
[00573] To a solution of ethyl 9'-hydroxy-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (110 mg, 0.3 mmol) in MeCN (1 mL) was added bromomethyl cyclopropane (61 mg, 0.45 mmol) and K2C03 (83 mg, 0.6 mmol). After stirred at 65 °C for 5 h, the reaction mixture was cooled down to room temperature, the solvents were removed under vacuum and the residue was purified by PTLC to provide the desired product (89 mg, 70% yield). LCMS (ESI) m/z: 422.4 (M + 1)+.
[00574] Step 2: 9'-(Cvclopropylmethoxy)-10'-methyl-2'-oxo-2'.7'- dihvdrospiro[cvclohexane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000124_0003
[00575] To a solution of ethyl 9'-(cyclopropylmethoxy)-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (89 mg, 0.21 mmol) in THF (2 mL) was added LiOH (20 mg, 0.84 mmol) and H20 (0.5 ml). After stirred at room temperature for 2 h, the reaction mixture was neutralized. The solvents were removed and the residue was purified by reverse phase reverse phase HPLC (C18, 30-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (48 mg, 58 % yield) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 8.80 (s, 1 H), 7.88 (s, 1 H), 7.29 (s, 1 H), 7.07 (s, 1 H), 3.95 (d, J=6.8 Hz, 2H), 3.26 (s, 2H), 2.22(s, 3H), 1.98-1 .89 (m, 2H), 1.80-1 .73 (m, 2H), 1 .70-1 .61 (m, 3H), 1 .59-1 .50 (m, 2H), 1 .36-1 .27 (m, 2H), 0.64-0.56 (m, 2H), 0.41 -0.33 (m, 2H). LCMS (ESI): 394.3 (M + 1)+.
[00576] Example 30
[00577] 10'-Methyl-2'-oxo-9'-(pyrrolidin-1 -yl)-2',7'-dihvdrospiro[cyclohexane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000125_0001
[00578] Stepl : Ethyl 10'-methyl-2'-oxo-9'-(pyrrolidin-1 -yl)-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate
Figure imgf000125_0002
[00579] To a mixture of ethyl 10'-methyl-2'-oxo-9'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate
[00580] (1 10 mg, 0.220 mmol), Pd2(dba)3 (20.1 mg,0.022mmol), BINAP (20.5 mg,0.033mmol), Cs2C03 (433.6 mg,1 .32mmol) in toluene (10 ml) was added pyrrolidine (31 .3 mg,0.44mmol). The resulting mixture was stirred at 100 °C for 5 h. Then the mixture was diluted with EtOAc (20ml_) and H20 (30 mL), and then extracted with EtOAc (50ml_x2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel, 0-10%MeOH in DCM) to afford ethyl 10'-methyl-2'-oxo-9'-(pyrrolidin-1 -yl)-2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (40 mg, 43.2% yield). LCMS (ESI) m/z: 421 .3 (M + 1)+.
[00581] Step 2: 10'-Methyl-2'-oxo-9'-(pyrrolidin-1 -yl)-2',7'-dihvdrospiro[cyclohexane-
Figure imgf000125_0003
[00582] To a solution of ethyl 10'-methyl-2'-oxo-9'-(pyrrolidin-1 -yl)-2',7'- dihydrospiro [cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (40 mg, 0.095 mmol) in MeOH (5 ml), NaOH (30.4 mg, 0.761 mmol) dissolved in H20 (1 ml) was added. The resulting mixture was stirred at room temperature for 2 h. The reaction mixture was acidified to ph~5 with 1 N HCI. Then the mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50ml_x2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by reverse phase reverse phase HPLC Gilson (C18 column, 30%~100%MeCN in H20, with 0.1 % formic acid in H20) to give 10'-methyl-2'-oxo-9'-(pyrrolidin-1 -yl)-2',7'-dihydrospiro[cyclohexane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (14 mg, 37.5% yield) as a yellow solid. LCMS (ESI) m/z: 393.8 (M + 1)+. Ή NMR (400 MHz, DMSO-d6) δ 8.77 (s, 1 H), 7.71 (s, 1 H), 7.20 (s, 1 H), 6.73 (s, 1 H), 3.40 (t, J = 6.4 Hz, 4H), 3.19 (s, 2H), 2.39 (s, 3H), 1 .93 - 1 .87 (m, 5H), 1 .82 - 1 .77 (m, 2H), 1 .69 - 1 .62 (m, 3H), 1 .60 - 1 .44 (m, 3H), 1 .32 - 1 .28 (m, 1 H).
[00583] Example 31
[00584] 9'-(Cvclopropylmethoxy)-10'-(difluoromethyl)-2'-oxo-2'.7'- dihvdrospiro[cvclohexa -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000126_0001
[00585] Step 1 : Ethyl 10'-bromo-9'-hvdroxy-2'-oxo-2',7'-dihvdrospiro[cyclohexane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000126_0002
[00586] A solution of ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (300 mg, 0.57 mmol) in TFA (8 ml_) was heated at 70 °C for 3 h. The solvents were concentrated. The mixture was diluted with DCM (30 ml_) and washed with brine (20 ml_ x 2). The organic layer was dried over Na2S04. The solvents were concentrated. The residue was triturated with PE/EA to afford the title compound (235 mg, 95% yield). LCMS (ESI) m/z: 432.3/434.1 (M/M+2)+. [00587] Step 2: Ethyl 10'-bromo-9'-(cvclopropylmethoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclohexane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000127_0001
[00588] To a solution of ethyl 10'-bromo-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (235 mg, 0.54 mmol) in DMF (5 mL) was added (bromomethyl)cyclopropane (146 mg, 1 .08 mmol) at room temperature. The reaction mixture was heated at 80 °C for 4 h. Water (20 mL) was added and the mixture was extracted with DCM (25 X 2 mL). The combined organic layers were washed with brine (30 X 4 mL). The solvents were concentrated. The residue was purified by column chromatography using MeOH in DCM (1/10) to afford the title compound (250 mg, 94% yield). LCMS (ESI) m/z: 486.4/488.3 (M/M+2)+.
[00589] Step 3: Ethyl 9'-(cvclopropylmethoxy)-2'-oxo-10'-vinyl-2'.7'- dihydrospirofcyclohexa -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000127_0002
[00590] To a solution of ethyl 10'-bromo-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (250 mg, 0.51 mmol) in EtOH (8 mL) was added potassium vinyl trifluoroborate (137 mg, 1 .02 mmol), Pd(PPh3)4 (58 mg, 0.05 mmol) and Na2C03 (162 mg, 1 .53 mmol) under N2 atmosphere. The reaction mixture was heated at 80 °C overnight. After cooling to rt, the mixture was filtered and washed with DCM (20 mL). The filtrated was concentrated and the residue was purified by column chromatography using EA/PE (30%-100%) to afford the title compound (188 mg, 84% yield). LCMS (ESI) m/z: 434.3 (M + 1)+. [00591] Step 4: Ethyl 9'-(cvclopropylmethoxy)-10'-formyl-2'-oxo-2'.7'- dihvdrospiro[cvclohexane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000128_0001
[00592] To a solution of ethyl 9'-(cyclopropylmethoxy)-2'-oxo-10'-vinyl-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (188 mg, 0.43 mmol) in THF (6 mL) and H20 (1.5 mL) was added K20s04 2H20 (16 mg, 0.043 mmol) at 0 °C, then Nal04 (368 mg, 1 .72 mmol) was added in portions. The reaction mixture was stirred at rt for 2 h. After cooling to rt, the mixture was filtered, washed with DCM (25 mL). The filtrated was concentrated and the residue was purified by prep-TLC using MeOH in DCM (1/15) to afford the title compound (145 mg, 77% yield). LCMS (ESI) m/z: 436.5 (M + 1)+.
[00593] Step 5: Ethyl 9'-(cvclopropylmethoxy)-10'-(difluoromethyl)-2'-oxo-2'.7'- dihydrospirofcyclohexane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000128_0002
[00594] To a solution of ethyl 9'-(cyclopropylmethoxy)-10'-formyl-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (145 mg, 0.33 mmol) in DCM (4 mL) was added dropwise DAST (213 mg, 1 .32 mmol) at 0 °C under N2 atmosphere. The reaction mixture was stirred at rt for overnight. The mixture was diluted with DCM (30 mL), washed with sat.NaHC03 solution (20 mL) and brine (20 mL). The solvent was concentrated to afford the title compound (140 mg, 92% yield). LCMS (ESI) m/z: 458.3 (M + 1)+.
[00595] Step 6: 9'-(Cvclopropylmethoxy)-10'-(difluoromethyl)-2'-oxo-2'.7'- dihvdrospiro[cvclohexa -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000128_0003
[00596] To a solution of ethyl 9'-(cyclopropylmethoxy)-10'-(difluoromethyl)-2'- oxo-2', 7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2, 1 -a]isoquinoline]-3'-carboxylate ( 140 mg, 0.31 mmol) in EtOH (6 mL) was added NaOH (53 mg, 1 .32 mmol) in H20 (1 .2 mL). The reaction mixture was stirred at rt for 2 h. The solvent was concentrated. The mixture was acidified to pH~1 by 1 N HCI and extracted with DCM (10 mL X 3). The combined organic layers were washed with brine (30 mL) and dried over Na2S04. The solvents were concentrated. The residue was purified by prep, reverse phase HPLC (C18, 30- 100 % acetonitrile in H20 with 0.1 % formic acid) to afford the title compound (30 mg, 32% yield) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 8.84 (s, 1 H), 8.14 (s, 1 H), 7.37 (s, 1 H), 7.35 (s, 1 H), 7.09 (t, J = 54.8 Hz, 1 H), 4.06 (d, J = 6.9 Hz, 2H), 3.37 (s, 2H), 2.02 - 1 .92 (m, J = 12.2, 9.1 Hz, 2H), 1 .80 - 1 .51 (m, 7H), 1 .38 - 1 .26 (m, 2H), 0.68 - 0.56 (m, 2H), 0.43 - 0.33 (m, 2H). LCMS (ESI) m/z: 430.3 (M + 1 )+.
[00597] Example 32
[00598] 9-(Benzyloxy)-10-bromo-6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H- pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000129_0001
[00599] Step 1 : Ethyl 2.3-dimethylbutanoate
Figure imgf000129_0002
[00600] To a freshly prepared LDA solution in THF (2.3 N, 260 mL) was added ethyl 3-methylbutanoate (65.1 g, 0.5 mol) dropwise at -78 °C. The resulting mixture was stirred at -78 °C for 1 .5 h before Mel (106.5 g, 0.75 mol) was added dropwise. The reaction mixture was stirred at -78 °C for 0.5 h and at rt for 2 h, followed by addition of H2O (300 mL) to quench the reaction. The resulting mixture was extracted with Et20 (4x200 mL), and the combined organic layers were washed with 1 N HCI (100 mL), NaHC03 (sat. aq., 100 mL) and brine. The organic phase was dried over Na2S04, filtered and concentrated to give the crude product (68.9 g, 96% crude yield) which was used in the next step without further purification. Ή NMR (400 MHz, CDCI3) δ 4.13 (q, J = 7.2 Hz, 2H), 1 .98 - 1 .83 (m, 1 H), 1 .25 (t, J = 7.2 Hz, 3H), 1 .21 (t, J = 6.8 Hz, 1 H), 1 .10 (d, J = 7.2 Hz, 3H), 0.99
(overlapped dd, J = 7.2 Hz, 6H). [00601] Step 2: Benzyl 3-(benzyloxy)-4-bromobenzoate
Figure imgf000130_0001
[00602] To a mixture of 4-bromo-3-hydroxybenzoic acid (90 g, 0.42 mol) and K2C03 (173 g, 1 .26 mol) in MeCN (1 .8 L) was added BnBr (142 g, 0.84mol) dropwise at room temperature over 0.5 h. The resulting mixture was stirred at 65 °C for 12 h before it was cooled down to room temperature. H20 (0.9 L) was added to quench the reaction, and the obtained mixture was extracted with EtOAc (2 x0.9 L). The combined organic layers were washed with brine, dried over MgS04 and concentrated under reduced pressure. The obtained residue was purified by recrystallization with MeOH (1 .8 L) to afford the title compound (140 g, 85% yield). LCMS (ESI) m/z: 397.4/399.4 (M/M+2)+.
[00603] Step 3: -(Benzyloxy)-4-bromophenyl) methanol
Figure imgf000130_0002
[00604] To a suspension of LiAIH4 (13.8 g, 0.36 mol) in THF (130 mL) was added the solution of benzyl 3-(benzyloxy)-4-bromobenzoate (144 g, 0.36 mol) in THF (1500 mL) dropwise at -20 °C over 2 h. The reaction mixture was stirred at -20 °C for 2 h before H20 (14 mL) was added dropwise over 0.5 h at 0 °C to quench the reaction. To the resulting mixture 40% NaOH (14 mL) was added, and the obtained suspension was stirred for 0.5 h before it was filtered with a Buchner funnel. The collected filtrate was concentrated under reduced pressure, and the obtained residue was dissolved in EtOAc (1500 mL). The solution was washed with brine, dried over MgS04 and concentrated to give the crude product which was purified by recrystallization with petroleum ether (1500 mL) to afford the title compound (72 g, 68% yield). 1 H NMR (400 MHz, DMSO-d6) δ 7.51 (dd, J = 12.9, 7.6 Hz, 3H), 7.42 (dd, J = 10.1 , 4.7 Hz, 2H), 7.36 - 7.32 (m, 1 H), 7.17 (d, J = 1 .4 Hz, 1 H), 6.90 - 6.80 (m, 1 H), 5.29 (t, J = 5.7 Hz, 1 H), 5.19 (s, 2H), 4.47 (t, J = 5.7 Hz, 2H).
[00605] Step 4: -(Benzyloxy)-1 -bromo-4-(bromomethyl) benzene
Figure imgf000130_0003
[00606] A solution of PPh3 (109.2 g, 0.416 mol) in THF (160 mL) was added dropwise to a mixture of (3-(benzyloxy)-4-bromophenyl)methanol (93.8 g, 0.32 mol) and CBr4 (138 g, 0.416 mmol) in THF (160 mL) at 0 °C. The resulting mixture was stirred at rt for 3 h before it was diluted with Et20 (600 mL). The suspension was filtered through a pad of celite, and the filtrate was concentrated to give the crude product which was purified by flash column chromatography (silica gel, 0-6% CH2CI2 in petroleum ether) to afford the title compound (87.8 g, 77% yield). Ή NMR (400 MHz, CDCI3) δ 7.54 - 7.45 (m, 3H), 7.44 - 7.37 (m, 2H), 7.37 - 7.30 (m, 1 H), 6.98 (d, J = 2.0 Hz, 1 H), 6.88 (dd, J = 8.4, 2.0 Hz, 1 H), 5.17 (s, 2H), 4.42 (s, 2H).
[00607] Step 5:
Figure imgf000131_0001
[00608] To a freshly prepared LDA solution in THF (2.4 N, 152 mL) was added the solution of ethyl 2,3-dimethylbutanoate (52.3 g, 0.363 mol) in THF (150 mL) dropwise at -78 °C. The resulting mixture was stirred at -78 °C for 1 .5 h before the solution of 2-(benzyloxy)-1 -bromo-4-(bromomethyl)benzene (80.7 g, 0.227 mol) in THF (150 mL) was added dropwise. The reaction mixture was allowed to warm up to rt slowly and stirred for 12 h. NH4CI solution (sat. aq., 300 mL) was added to quench the reaction, and the mixture was extracted with EtOAc (4x200 mL). The combined organic layers were washed with 1 N HCI (100 mL), NaHC03 (sat. aq., 100 mL) and brine. The organic phase was dried over Na2S04, filtered and concentrated to give the crude product which was purified by flash column chromatography (silica gel, 0-5% Et20 in petroleum ether) to afford the title compound (84.3 g, 88% yield). 1H NMR (400 MHz, CDCI3) δ 7.50 -7.44 (m, 2H), 7.42 - 7.35 (m, 3H), 7.34 - 7.27 (m, 1 H), 6.70 (d, J = 2.0 Hz, 1 H), 6.59 (dd, J = 8.0, 2.0 Hz, 1 H), 5.10 (s, 2H), 4.02 (qd, J = 6.8, 2.4 Hz, 2H), 3.02 (d, J = 12.8 Hz, 1 H), 2.55 (d, J = 12.8 Hz, 1 H), 2.05 (p, J = 6.8 Hz, 1 H), 1 .19 (t, J = 6.8 Hz, 3H), 0.94 (d, J = 6.8 Hz, 3H), 0.89 (s, 3H), 0.87 (d, J = 6.8 Hz, 3H).
[00609] Step 6: -(3-(Benzyloxy)-4-bromobenzyl)-2,3-dimethylbutanoic acid
Figure imgf000131_0002
[00610] To a solution of ethyl 2-(3-(benzyloxy)-4-bromobenzyl)-2,3- dimethylbutanoate (84.3 g, 0.201 mol) in DMSO/H2O (201 mL/10 mL) was added KOH (33.8 g, 0.603 mol). The resulting mixture was stirred at 120 °C for 16 h, cooled down to rt and then diluted with EtOAc (1000 mL). The obtained mixture was acidified with 1 N HCI to pH2. The organic phase was separated, washed with H20 (4100 mL), dried over Na2S04 and concentrated to provide 79.9 g crude material which was used directly in the next step without further purification. Ή NMR (400 MHz, CDCI3) δ 7.49 - 7.42 (m, 2H), 7.41 - 7.33 (m, 3H), 7.33 - 7.27 (m, 1 H), 6.78 (d, J = 1 .6 Hz, 1 H), 6.65 (dd, J = 8.0, 2.0 Hz, 1 H), 5.10 (s, 2H), 2.99 (d, J = 13.2 Hz, 1 H), 2.58 (d, J = 13.2 Hz, 1 H), 2.05 (dt, J = 14.0, 6.8 Hz, 1 H), 0.95 (d, J = 6.8 Hz, 3H), 0.90 (d, J = 6.8 Hz, 3H), 0.88 (s, 3H).
[00611] Step 7: -(Benzyloxy')-1 -bromo-4-(2-isocvanato-2,3-dimethylbutyl') benzene
Figure imgf000132_0001
[00612] To a mixture of 2-(3-(benzyloxy)-4-bromobenzyl)-2,3- dimethylbutanoic acid (79.9 g, 0.204 mol) from previous step and Et3N (24.8 g, 0.245 mol) in PhMe (204 mL) was added DPPA (67.4 g, 0.245 mol) dropwise. The resulting mixture was stirred at 120 °C for 3 h before it was cooled down to room temperature. The reaction mixture was concentrated to provide 172 g crude material which was used directly in the next step immediately. LCMS (ESI) m/z: 388.37/391 .31 (M/M+2)+.
[00613] Step 8: -(3-(Benzyloxy)-4-bromophenyl)-2,3-dimethylbutan-2-amine
Figure imgf000132_0002
[00614] To a solution of 2-(benzyloxy)-1 -bromo-4-(2-isocyanato-2,3- dimethylbutyl)benzene 172 g from previous step in THF (204 mL) was added HCI (cone. aq. 180 mL) dropwise at 0 °C. The resulting mixture was allowed to warm up to rt and stirred for 12 h, and then basified with 5 N NaOH to pH around 12. The mixture was extracted with EtOAc (4 x300 mL). The combined organic layers were washed with H2O (2x300 mL) and brine, dried over Na2S04, filtered and concentrated to provide 72.3 g crude 1 -(3- (Benzyloxy)-4-bromophenyl)-2,3-dimethylbutan-2-amine which was used in the next step without further purification. LCMS (ESI) m/z: 362.2/364.2 (M/M+2)+.
[00615] Step 9: N-(1 -(3-(benzyloxy)-4-bromophenyl)-2,3-dimethylbutan-2- vDformamide
Figure imgf000132_0003
[00616] A mixture of formic acid (16.2 g, 0.352 mol) and acetic anhydride
(32.6 g, 0.32 mol) was stirred at 60 °C for 1 .5 h before it was cooled down to room temperature. This freshly prepared acetic formic anhydride was added dropwise to a mixture of 1 -(3-(benzyloxy)-4-bromophenyl)-2,3-dimethylbutan-2-amine (72.3 g, 0.2 mol) and Et3N (40.5 g, 0.4 mol) in DCM (200 mL) at 0 °C. The resulting mixture was allowed to warm up to rt and stirred for 12 h before H20 (100 mL) was added to quench the reaction. The organic phase was separated, washed with H20 (100ml_) and NaHC03 (sat. aq., 200 mL), dried over Na2S04, filtered and concentrated to provide 70.0 g crude N-(1 -(3- (benzyloxy)-4-bromophenyl)-2,3-dimethylbutan-2-yl)formamide which was used in the next step without further purification. LCMS (ESI) m/z: 390.2/392.2 (M/M+2)+.
[00617] Step 10: 6-(Benzyloxy)-7-bromo-3-isopropyl-3-methyl-3,4- dihvdroisoquinoline
Figure imgf000133_0001
[00618] To a solution of N-(1 -(3-(benzyloxy)-4-bromophenyl)-2,3- dimethylbutan-2-yl)formamide (70.0 g, 0.179 mol) in CH2CI2 (358 mL) was added POCI3 (28.8 g, 0.188 mol) dropwise at 0 °C. The resulting mixture was allowed to warm up to rt and stirred for 14 h before it was basified with 2 N NaOH to pH around 12. The organic phase was isolated, and the aqueous phase was extracted with EtOAc (4x250 mL). The combined organic layers were washed with H20 (2x300 mL) and brine, dried over Na2S04, filtered and concentrated to provide 66.0 g crude 6-(Benzyloxy)-7-bromo-3-isopropyl-3-methyl-3,4- dihydroisoquinoline which was used in the next step without further purification. LCMS (ESI) m/z: 372.2/374.2 (M/M+2)+.
[00619] Step 1 1 : Ethyl 9-(benzyloxy)-10-bromo-6-isopropyl-6-methyl-2-oxo- 1 ,6,7,1 1 b-tetrahvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000133_0002
[00620] A mixture of 6-(benzyloxy)-7-bromo-3-isopropyl-3-methyl-3,4- dihydroisoquinoline (66.0 g) and ethyl 2-(ethoxymethylene)-3-oxobutanoate (164.8 g, 0.885 mol) in EtOH (177 mL) was stirred at 100 °C for 20 h. After cooling down to rt, the reaction mixture was concentrated and was purified by flash column chromatography (silica gel, CH2CI2: MeOH : NH4OH = 800 : 8 : 1 ) to afford the title compound (21 .2 g, 21 % yield over 6 steps) as a pale brown solid. LCMS (ESI) m/z: 512.2/514.2 (M/M+2)+. [00621] Step 12: Ethyl 9-(benzyloxy)-10-bromo-6-isopropyl-6-methyl-2-oxo-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000134_0001
[00622] A mixture of ethyl 9-(benzyloxy)-10-bromo-6-isopropyl-6-methyl-2- oxo-1 ,6,7,1 1 b-tetrahydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (21 .2 g, 41 .4 mmol) and p-chloranil (8.7 g, 35.2 mmol) in DME (166 mL) was stirred at 80 °C for 12 h. After cooling to rt, the resulting suspension was filtered with a Biichner funnel. The collected insoluble solid was washed with cold DME (3x4 mL) and then dried under high vacuum to afford the title compound (16.8 g, 80% yield) as a slightly brown solid. 1H NMR (400 MHz, CDC ) δ 8.40 (s, 1 H), 7.92 (s, 1 H), 7.48 - 7.43 (m, 2H), 7.43 - 7.37 (m, 2H), 7.37 - 7.31 (m, 1 H), 6.90 (s, 1 H), 6.74 (s, 1 H), 5.22 (s, 2H), 4.39 (q, J = 7.2 Hz, 2H), 3.08 (d, J = 16 Hz, 1 H), 2.98 (d, J = 16 Hz, 1 H) 1 .86 (p, J = 6.8 Hz, 1 H), 1 .61 (s, 3H), 1 .40 (t, J = 7.2 Hz, 3H), 0.78 (d, J = 6.8 Hz, 3H), 0.72 (d, J = 6.8 Hz, 3H). LCMS (ESI) m/z: 510.2/512.3 (M/M+2)+.
[00623] Step 13: 9-(Benzyloxy)-10-bromo-6-isopropyl-6-methyl-2-oxo-6,7-dihydro- 2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000134_0002
[00624] To a solution of ethyl 9-(benzyloxy)-10-bromo-6-isopropyl-6-methyl-
2-0X0-6, 7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (61 mg, 0.12 mmol) in THF/H20 (0.6 ml/0.6 mL) was added LiOH (12 mg, 0.48 mmol). After stirring at rt for 2 h, the resulting mixture was acidified with 1 N HCI and extracted with CH2CI2 (3 x5 mL). The combined organic layers were washed with brine, dried over Na2S04, filtered and concentrated to provide the crude product which was purified by reverse phase reverse phase HPLC (C18, 0-90% MeCN in H20 with 0.1 % formic acid) to afford the title compound (23.7 mg, 41 % yield) as an off-white powder. Ή NMR (400 MHz, DMSO-d6) δ 8.55 (s, 1 H), 8.34 (s, 1 H), 7.54 - 7.48 (m, 2H), 7.47 - 7.39 (m, 3H), 7.39 - 7.31 (m, 2H), 5.36 - 5.28 (m, 2H), 3.30 - 3.18 (m, 2H), 1 .77 (dt, J = 13.6, 6.8 Hz, 1 H), 1 .67 (s, 3H), 0.78 (d, J = 6.8 Hz, 3H), 0.62 (d, J = 6.8 Hz, 3H). LCMS (ESI) m/z: 482.2/484.2 (M/M+2)+.
[00625] Example 33 [00626] 10-Cvclopropyl-6-isopropyl-6-methyl-9-(1 -methyl-1 H-pyrazol-4-vD-2-oxo-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000135_0001
[00627] Step 1 : Ethyl 9-(benzyloxy)-10-cvclopropyl-6-isopropyl-6-methyl-2-oxo-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000135_0002
[00628] Under a stream of nitrogen, ethyl 9-(benzyloxy)-10-bromo-6- isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (500 mg, 1 .0 mmol), cyclopropylboronic acid (170 mg, 2.0 mmol), Pd(Ph3)4 (46 mg, 0.04 mmol), Cs2C03 (1 .3 g, 4.0 mmol), toluene (10 ml_) and water (2 ml_) were placed into a Single round bottom flask, then stirred overnight at 100 °C. The reaction mixture was cooled, added to water (100 ml_), and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, and was concentrated to give an oily liquid. This crude material was dissolved in a small amount of DCM, packed to silica gel chromatography, and subjected to column purification using DCM/MeOH as an eluent to afford the title compound (330 mg, 70% yield), LCMS (ESI) m/z: 472.3 (M + 1 )+.
[00629] Step 2: Ethyl 10-cvclopropyl-9-hvdroxy-6-isopropyl-6-methyl-2-oxo-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000135_0003
[00630] A solution of ethyl 9-(benzyloxy)-10-cyclopropyl-6-isopropyl-6- methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (330 mg, 0.66 mmol) in TFA (5 ml_) was stirred at 65 °C. After the reaction was finished, the mixture was concentrated. The residue was dissolved in DCM, washed with saturated NaHC03 aqueous, dried over MgS04 and concentrated. The crude material was purified with flask chromatography to afford the title compound (181 mg, 72% yield). LCMS (ESI) m/z: 382.2 (M + 1 )+.
[00631] Step 3: Ethyl 10-cvclopropyl-6-isopropyl-6-methyl-2-oxo-9- (((trifluoromethyl)sulfonyl)oxy)-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000136_0001
[00632] To a solution of ethyl 10-cyclopropyl-9-hydroxy-6-isopropyl-6-methyl-
2-0X0-6, 7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (180 mg, 0.47 mmol) in CH3CN (2 mL) was added PhN(Tf)2 (201 mg, 0.56 mmol), DIPEA (182 mg, 1 .41 mmol) and DMAP (2.9 mg, 0.024 mmol). The reaction mixture was stirred at room temperature for 2 h. Then the mixture was filtered, and the residue was washed with DCM. The filtrate was evaporated. The crude product was purified with flash column (20 g silica gel. Solvent: MeOH/ DCM, 1 : 10) to afford the title compound (102 mg, 42% yield). LCMS (ESI) m/z: 514.2 (M + 1)+.
[00633] Step 4: Ethyl 10-cvclopropyl-6-isopropyl-6-methyl-9-(1 -methyl-1 H-pyrazol-4- yl)-2-oxo-6,7-dihvdro-2 -pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000136_0002
[00634] Under a stream of nitrogen, ethyl 10-cyclopropyl-6-isopropyl-6- methyl-2-oxo-9-(((trifluoromethyl)sulfonyl)oxy)-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3- carboxylate (100 mg, 0.2 mmol), 1 -methyl-4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)- 1 H-pyrazole (50 mg, 0.24 mmol), Pd(PPh3)4 (46 mg, 0.04 mmol), Cs2C03 (130 mg, 0.4 mmol), toluene (2 mL) and water (1 mL) were placed into a Schlenk tube, then stirred overnight at 100 °C. The reaction mixture was added to water (20 mL), and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure, and an oily liquid was obtained. This was dissolved in a small amount of DCM, packed to a silica gel chromatography and subjected to column purified using DCM/ MeOH as an eluent, to afford the title compound (58 mg, 65% yield). LCMS (ESI) m/z: 446.3 (M + 1)+. [00635] Step 5: 10-Cvclopropyl-6-isopropyl-6-methyl-9-(1 -methyl- 1 H-pyrazol-4-yl)-2- ,7-dihvdro-2H-pyr -alisoquinoline-3-carboxylic acid
Figure imgf000137_0001
[00636] To a solution of ethyl 10-cyclopropyl-6-isopropyl-6-methyl-9-(1 - methyl-1 H-pyrazol-4-yl)-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (15 mg, 0.03 mmol) in MeOH (2 mL) and water (1 mL) was added LiOH (1 .2 mg, 0.05 mmol). The reaction mixture was stirred at room temperature for 2 h, The mixture was acidified to pH~3 and purified by reverse phase reverse phase HPLC (C18 0-40% MeCN in H20 with 0.1 % formic acid) to afford the title compound (2 mg, 16% yield) as a white solid. LCMS (ESI) m/z: 418.9 (M + 1)+.Ή NMR (400 MHz, DMSO-d6) δ 8.58 (s, 1 H), 8.12 (s, 1 H), 7.97 (s, 1 H), 7.68 (s, 1 H), 7.62 (s, 1 H), 7.42 (s, 1 H), 3.92 (s, 3H), 3.27 - 3.22 (m, J = 9.5 Hz, 2H), 2.02 -1 .97 (m, 1 H), 1 .90 (d, J = 5.4 Hz, 2H), 1 .83 - 1 .78 (m, 1 H), 1 .69 (s, 3H), 0.92 - 0.76 (m, J = 10.3 Hz, 4H), 0.63 (d, J = 6.8 Hz, 3H).
[00637] Example 34
[00638] 9-(Cvclopropylmethoxy)-10-(difluoromethyl)-6-isopropyl-6-methyl-2-oxo-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000137_0002
[00639] Step 1 : Ethyl 10-bromo-9-hvdroxy-6-isopropyl-6-methyl-2-oxo-6,7-dihydro- 2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000137_0003
[00640] A solution of ethyl 9-(benzyloxy)-10-bromo-6-isopropyl-6-methyl-2- oxo-6, 7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (840 mg, 1 .65 mmol) in TFA (40 mL) was stirred at 70 °C. The reaction mixture was concentrated. The residue was dissolved in DCM, washed with K2C03, dried over MgS04 and concentrated. The crude material was purified with flash chromatography to afford the title compound (580 mg, 84% yield). LCMS (ESI) m/z: 420.1/422.1 (M/M+2)+.
[00641] Step 2: Ethyl 10-bromo-9-(cvclopropylmethoxyy6-isopropyl-6-methyl-2-oxo- 6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000138_0001
[00642] To a solution of ethyl 10-bromo-9-hydroxy-6-isopropyl-6-methyl-2- oxo-6, 7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (350 mg, 0.83 mmol) in DMF (5 mL) was added K2C03 (229 mg, 1 .67 mmol) and (bromomethyl)cyclopropane (167 mg, 1 .24 mmol). After stirred at 80 °C for 72 h, the reaction mixture was cooled to room temperature, quenched with H20, and was extracted with ethyl acetate three times. The combined organic layers were washed with brine and dried over Na2S04. The solvent was removed under reduced pressure, and the crude material was subjected to silica gel column purification to afford the title compound (310 mg, 79% yield). LCMS (ESI) m/z: 474.2/474.2 (M/M+2)+.
[00643] Step 3: Ethyl 9-(cvclopropylmethoxyy6-isopropyl-6-methyl-2-oxo-10-vinyl- 6,7-dihvdro-2H-pyrido[ -alisoquinoline-3-carboxylate
Figure imgf000138_0002
[00644] Under a stream of nitrogen, ethyl 10-bromo-9-(cyclopropylmethoxy)-
6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (350 mg, 0.74 mmol), 4,4,5,5-tetramethyl-2-vinyl-1 ,3,2-dioxaborolane (231 mg, 1 .5 mmol), Pd(Ph3)4 (172 mg, 0.15 mmol), KOAc (132 mg, 1 .5 mmol), dixoane (5 mL) and water (1 mL) were added into a Schlenk tube, then stirred overnight at 100 °C. The reaction mixture was diluted with 40 mL water, and extracted with DCM. The organic layer was dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The crude material was subjected to column purification using DCM/MeOH as an eluent to afford the title compound (250 mg, 79% yield), LCMS (ESI) m/z: 422.4 (M + 1)+. [00645] Step 4: Ethyl 9-(cvclopropylmethoxy)-10-formyl-6-isopropyl-6-methyl-2-oxo-
Figure imgf000139_0001
[00646] Ethyl 9-(cyclopropylmethoxy)-6-isopropyl-6-methyl-2-oxo-10-vinyl-
6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (250 mg, 0.59 mmol), Nal04 (505 mg, 2.36 mmol) and distilled water were sequentially added to a water (5 mL) solution of the K2040s (9.9 mg, 0.03 mmol) under an inert atmosphere and the mixture was stirred at 25 °C. The reaction mixture was extracted with DCM, concentrated and the residue was purified by column chromatography on silica gel (DCM: MeOH) to afford the title compound (220 mg, 88% yield). LCMS (ESI) m/z: 424.2 (M + 1 )+.
[00647] Step 5: Ethyl 9-(cyclopropylmethoxy)-10-(difluoromethyl)-6-isopropyl-6- methyl-2-oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000139_0002
[00648] To a solution of ethyl 9-(cyclopropylmethoxy)-10-formyl-6-isopropyl-
6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (220 mg, 0.52 mmol) in DCM (3 mL) was added a solution of DAST (418 mg, 2.6 mmol) in DCM (2 mL) dropwise over 30 min at 0 °C. The mixture was poured into saturated NaHC03 aqueous at 0 °C and was extracted twice with DCM. The combined DCM layers were washed with brine, dried over MgS04 and concentrated to afford the title compound (140 mg, 60% yield). LCMS (ESI) m/z: 446.4 (M + 1)+.
[00649] Step 6: ^(CvclopropylmethoxyVI O^difluoromethvO-e-isopropyl-e-methyl^- oxo-6, 7-dihydro-2H-pyr -alisoquinoline-3-carboxylic acid
Figure imgf000140_0001
[00650] To a solution of ethyl 9-(cyclopropylmethoxy)-10-(difluoromethyl)-6- isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (140 mg, 0.31 mmol) in EtOH (50 mL) was added NaOH (62 mg, 1 .55 mmol) at room temperature After stirred at room temperature for 2 h, the reaction mixture was acidified to pH~1 and extracted with ethyl acatate three times. The combined organic layers were washed with brine and dried over Na2S04. Solvent were removed and the residue was purified by reverse phase reverse phase HPLC (C18 0-40% MeCN in H20 with 0.1 % formic acid) to afford the title compound (108 mg, 83% yield) as a white solid. LCMS (ESI) m/z: 418.2 (M + 1)+ . Ή NMR (400 MHz, DMSO-d6) δ 8.56 (s, 1 H), 8.15 (s, 1 H), 7.40 (s, 1 H), 7.27 (s, 1 H), 7.22 (s, 0.25H), 7.08 (s, 0.5H), 6.94 (s, 0.25H), 4.06 (d, J = 4.8 Hz, 2H), 3.24 (s,2H), 1 .78 - 1 .74 (m, 1 H), 1 .68 (s, 3H), 0.88 - 0.83 (m, 1 H), 0.81 (d, J = 6.8 Hz, 3H), 0.63 (d, J = 6.8 Hz, 3H), 0.61 - 0.58 (m, 2H), 0.40 - 0.37 (m, 2H).
[00651] Example 35
[00652] Enantiomer 1 : 9-(Cyclopropylmethoxy)-10-(difluoromethyl)-6-isopropyl-6- methyl-2-oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
[00653] Example 36
[00654] Enantiomer 2: 9-(Cyclopropylmethoxy)-10-(difluoromethyl)-6-isopropyl-6- methyl-2-oxo-6, -dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000140_0002
[00655] Ethyl 9-(cyclopropylmethoxy)-10-(difluoromethyl)-6-isopropyl-6-methyl-2- oxo-6, 7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (75.8 mg) was subjected to chiral chromatography to give two enantiomers. The enantiomers were hydrolyzed independently in THF (2 mL), Water (0.5 mL) with LiOH (21 .1 mg) to give 18 mg ( 24.5 % yield) of Enantiomer 1 and 20 mg (27.3 % yield) of Enantiomer 2 respectively. [00656] Enantiomer 1 : Ή NMR (400 MHz, DMSO-d6) δ 8.56 (s, 1 H), 8.15 (s, 1 H), 7.40 (s, 1 H), 7.27 (s, 1 H), 7.22 (s, 0.25H), 7.08 (s, 0.5H), 6.94 (s, 0.25H), 4.06 (dd,
[00657] J = 6.9, 2.4 Hz, 2H), 3.32 (s, 2H), 1 .80 - 1 .74 (m, 1 H), 1 .68 (s, 3H), 1 .32 - 1 .27 (m, 1 H), 0.81 (d, J = 6.8 Hz, 3H), 0.63 (d, J = 6.8 Hz, 3H), 0.60 (dd, J = 8.0, 1 .6 Hz, 2H), 0.43 - 0.35 (m, 2H). (ESI) m/z: 418.5 (M + 1 )+
[00658] Enantiomer 2: Ή NMR (400 MHz, DMSO-d6) δ 8.56 (s, 1 H), 8.15 (s, 1 H), 7.40 (s, 1 H), 7.27 (s, 1 H), 7.22 (s, 0.25H), 7.08 (s, 0.5H), 6.94 (s, 0.25H), 4.06 (m, 2.3 Hz, 2H), 3.32 (s, 2H), 1 .82 - 1 .74 (m, 1 H), 1 .68 (s, 3H), 1 .28 (s, 1 H), 0.81 (d, J = 6.8 Hz, 4H), 0.63 (d, J = 6.8 Hz, 3H), 0.62 - 0.57 (m, 2H), 0.39 (d, J = 2.2 Hz, 2H). (ESI) m/z: 418.3 (M + D+
[00659] Example 37
[00660] 6-lsopropyl-10-methoxy-9-(3-methoxypropoxy)-6-methyl-2-oxo-6,7-dihydro- 2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000141_0001
[00661] Step 1 : 4-Methoxy-3-(3-methoxypropoxy)benzaldehvde
Figure imgf000141_0002
[00662] To a stirred solution of 3-hydroxy-4-methoxybenzaldehyde (40 g,
0.17 mol) in MeCN (500 mL ) was added 1 -bromo-3-methoxypropane (32 g, 0.21 mol) and K2C03 (70 g, 0.51 mol). The resulting mixture was refluxed overnight, before cooled down to room temperature and quenched with H20. The mixture was extracted with EtOAc (100ml_ three times) and the combined organic layers were washed with brine and dried over Na2S04. The solvents were removed to give 50 g of the crude product 4-methoxy-3-(3- methoxypropoxy)benzaldehyde as a white solid which was used in the next step without purification. LCMS (ESI) m/z: 225.3 (M + 1)+.
[00663] Step 2: (4-Methoxy-3-(3-methoxypropoxy)phenyl)methanol
Figure imgf000141_0003
[00664] To a 0 °C, solution of 4-methoxy-3-(3-methoxypropoxy)benzaldehyde
(50 g, 0.22 mol) in MeOH (500 mL) was added NaBH4(16 g, 0.44 mol) in portions. After stirring at 0 °C for 2h, the reaction mixture was concentrated and partitioned between EtOAc and H20. The organic layers were separated, washed with brine, dried over Na2S04 and concentrated to dryness to give 45 g crude as a white solid which was used in the next step without purification. LCMS (ESI) m/z: 227.1 (M + 1)+.
[00665] Step 3: -(bromomethyl)-1 -methoxy-2-(3-methoxypropoxy)benzene
Figure imgf000142_0001
[00666] TMS-Br (17.5 mL, 0.13 mol) was added dropwise to a 0°C cooled solution of (4-methoxy-3-(3-methoxypropoxy)phenyl)methanol (20 g, 0.088 mol) in CHCI3 (200 mL). The mixture was stirred at rt for 30 minutes and stood for 1 h. The upper liquid was collected and concentrated. The residue was purified by flash chromatography (silica gel, 0-15% EtOAc in petroleum ether) to afford the title compound (22 g, 86.3% yield) as a white solid. 1H NMR (400 MHz, CDCI3) δ 6.87 (m, 2H), 6.75 (d, J = 8.0 Hz, 1 H), 4.42 (s, 2H), 4.06 (t, J = 6.5 Hz, 2H), 3.79 (s, 3H), 4.06 (t, J = 6.1 Hz, 2H), 3.29 (s, 3H), 4.06 (p, J = 6.3 Hz, 2H).
[00667] Step 1 : Ethyl 2.3-dimethylbutanoate EtOOCN
[00668] To a freshly prepared solution of LDA (2.0 M, 156 mmol) in THF (35 mL) was added methyl ethyl 3-methylbutanoate (6 g, 130 mmol) at -78°C. The solution was stirred for 1 hour before Mel (7.8 mL, 18.5 g, 130 mol) was added. After another hour at - 78°C, the solution was allowed to warm to room temperature and stirred for 2 hours. The reaction was quenched with water (100 mL), extracted with Et20 (3- 150 mL), the organic layers were washed with saturated NH4CI (aq) (100 mL), brine and dried with Na2S04. After filtration, the solvent was removed under reduced pressure. The title compound was obtained as a colorless liquid (5.2 g) Ή NMR (400 MHz, CDCI3) δ 4.13 (q, J = 7.2 Hz, 2H), 1 .98 - 1 .83 (m, 1 H), 1 .25 (t, J = 7.2 Hz, 3H), 1 .21 (t, J = 6.8 Hz, 1 H), 1 .10 (d, J = 7.2 Hz, 3H), 0.99 (overlapped dd,J = 7.2 Hz, 6H).
[00669] Step 2: Ethyl 2-(4-methoxy-3-(3-methoxypropoxy)benzyl)-2,3- dimethylbutanoate
Figure imgf000142_0002
[00670] Ethyl 2,3-dimethylbutanoate (1 .7 g, 12 mmol) was added slowly to a solution of LDA (7.5 ml, 15 mmol) in THF (10 ml) at -78°C. The mixture was stirred for 1 h at -78°C before addition of 2-(benzyloxy)-4-(bromomethyl)-1 -methoxybenzene (3 g, 10 mmol) in solution of THF (20 ml) was added dropwise. The reaction mixture was stirred for 1 h at - 78°C and at room temperature for another 2h. After quenched with water, EtOAc was added to the mixture and the organic layer was separated, dried over MgS04, concentrated. The crude material was purified via silica gel chromatography (5-30% EtOAc/Hex) to give ethyl 2-(4-methoxy-3-(3-methoxypropoxy)benzyl)-2,3-dimethylbutanoate (3.1 g, 88 % yield) as a white solid. 1H NMR (400 MHz, CDCb) 66.74 (d, J = 8.1 Hz, 1 H), 6.68 - 6.60 (m, 2H), 4.12 - 4.03 (m, 4H), 3.82 (s, 3H), 3.56 (t, J = 6.2 Hz, 2H), 3.35 (s, 3H), 3.01 (d, J = 13.2 Hz,1 H), 2.56 (d, J = 13.2 Hz, 1 H), 2.13 - 2.03 (m, 3H), 1 .22 (t, J = 7.1 Hz, 3H), 0.98 - 0.95 (m, 6H), 0.89 - 0.87 (m, 3H).
[00671] Step 3: 2-(4-Methoxy-3-(3-methoxypropoxy)benzyl)-2,3-dimethylbutanoic acid
Figure imgf000143_0001
[00672] Ethyl 2-(4-methoxy-3-(3-methoxypropoxy)benzyl)-2,3- dimethylbutanoate(3 g, 8 mmol) from previous step was dissolved in DMSO (18 ml), Water (6 ml) and was treated with KOH (2.3 g, 40 mmol) at 120°C overnight. The reaction mixture was acidified with 1 N HCI to pH4, and the mixture was concentrated. Water and EtOAc were added and the organic layer was separated. The solvent was concentrated, and the crude material was purified via silica gel chromatography (2-10% EtOAc/Hex) to give 2-(4- methoxy-3-(3-methoxypropoxy) benzyl)-2,3-dimethylbutanoic acid (2.2 g, 79 % yield) as a white solid. LCMS (ESI) m/z: 325.2 (M + 1)+.
[00673] Step 4: 1 -(4-Methoxy-3-(3-methoxypropoxy)phenyl)-2,3-dimethylbutan-2- amine
Figure imgf000143_0002
[00674] A solution of 2-(4-methoxy-3-(3-methoxypropoxy)benzyl)-2,3- dimethylbutanoic acid (2.2 g, 6.7 mmol), DPPA (1 .43 mL, 6.7 mmol) and Et3N (0.90 mL, 6.7 mmol) in toluene (20 mL) in a sealed tube was purged with nitrogen, and stirred with heating at 90 oC for 5h. Cone. HCI (3 mL, 80 mmol) was added to the mixture at OoC and the reaction was stirred overnight at room temperature. The solvent was concentrated, and the crude material was purified via silica gel chromatography (2-10% MeOH/DCM) to give 1 -(4- methoxy-3-(3-methoxypropoxy) phenyl)-2,3-dimethylbutan-2-amine (1 .8 g, 89 % yield) as a white solid. LCMS (ESI) m/z: 296.1 (M + 1)+.
[00675] Step 5: N-(1 -(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,3-dimethylbutan-2- vDformamide
Figure imgf000144_0001
[00676] 1 -(4-Methoxy-3-(3-methoxypropoxy)phenyl)-2,3-dimethylbutan-2-amine (1 .8 g, 0.504 mmol) was dissolved in dioxane (20 mL) and was treated with a pre-mixed solution consisting of formic acid (5 mL, 130 mmol) and acetic anhydride (5 mL, 53.0 mmol) dissolved in dioxane (5.0 mL), and the mixture was heated at 90 oC with stirring for one hour. The reaction mixture was cooled to room temperature and the solvent was evaporated. Water and EtOAc were added and the organic layer was separated, and was dried over MgS04. The solvent was evaporated to give the crude N-(1 -(4-methoxy-3-(3- methoxypropoxy)phenyl)-2,3-dimethylbutan-2-yl)formamide (1 .2 g, 60 % yield) as a yellow oil, which was used directly in the next step without purification. LCMS (ESI) m/z: 324.6 (M + 1)+.
[00677] Step 5: 3-lsopropyl-7-methoxy-6-(3-methoxypropoxy)-3-methyl-3,4- dihvdroisoquinoline
Figure imgf000144_0002
[00678] N-(1 -(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,3-dimethylbutan-2- yl)formamide (1 .2 g, 3.7 mmol) was dissolved in acetonitrile (10 mL) and was heated with POCI3 (0.34 mL, 3.7 mmol) at 90 °C for one hour. The reaction mixture was cooled to 0 °C and was basified with NH4OH until pH>1 1 . EtOAc and water were added, and the organic layer was separated and was dried over MgS04. The solvent was evaporated to give 700 mg crude 3-isopropyl-7-methoxy-6-(3-methoxypropoxy)-3-methyl-3,4-dihydroisoquinoline as a dark solid, which was used in the next step without purification. LCMS (ESI) m/z: 306.2 (M
+ 1)+.
[00679] Step 5: Ethyl 6-isopropyl-10-methoxy-9-(3-methoxypropoxy)-6-methyl-2- oxo-1 ,6,7,1 1 b-tetrahvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000145_0001
[00680] The crude 3-isopropyl-7-methoxy-6-(3-methoxypropoxy)-3-methyl-3,4- dihydroisoquinoline (700 mg, 2.29 mmol) was dissolved in ethanol (10.00 ml_) and was treated with ethyl 2-(ethoxymethylene)-3-oxobutanoate (1 .26 g, 6.782 mmol) at 84 °C overnight and was then stirred for three more days at 100 °C to drive the reaction to completion. The reaction mixture was concentrated, and was diluted with water and EtOAc. The organic layer was separated, dried over MgS04 and was concentrated. The crude material was purified via silica gel chromatography (2-10% MeOH/DCM) to give ethyl 6- isopropyl-10-methoxy-9-(3-methoxypropoxy)-6-methyl-2-oxo-1 ,6,7,1 1 b-tetrahydro-2H- pyrido[2,1 -a]isoquinoline-3-carboxylate (550 mg, 61 % yield) as a yellow solid. LCMS (ESI) m/z: 446.4 (M + 1)+.
[00681] Step 5: Ethyl 6-isopropyl-10-methoxy-9-(3-methoxypropoxy)-6-methyl-2- oxo-6, 7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000145_0002
[00682] The ethyl 6-isopropyl-10-methoxy-9-(3-methoxypropoxy)-6-methyl-2-oxo- 1 ,6,7,1 1 b-tetrahydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (550 mg, 1 .24 mmol) was dissolved in DME (5 ml_) and was treated with 2,3,5,6-tetrachlorocyclohexa-2,5-diene-1 ,4- dione (243 mg, 0.99 mmol) for three hours at 80 °C. The reaction mixture was concentrated, then diluted with water and EtOAc, and the organic layer was separated. The organic layer was dried over MgS04 and the solvent was evaporated. The resulting residue was suspended in cold DME and a dark tan solid was filtered off to give 420 mg of crude title compound which was used directly in the next step without purification. LCMS (ESI) m/z: 444.6 (M + 1)+. [00683] Step 5: 6-lsopropyl-10-methoxy-9-(3-methoxypropoxy)-6-methyl-2-oxo-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000146_0001
[00684] The crude ethyl 6-isopropyl-10-methoxy-9-(3-methoxypropoxy)-6-methyl-2- oxo-6, 7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (100 mg, 0.225 mmol) from previous step was dissolved in THF (3 ml_), Water (1 ml_) and was treated with LiOH (27 mg, 1 .12 mmol) at room temperature for 30 minutes. The reaction mixture was acidified with 1 N HCI to pH4, and the mixture was concentrated. Water and EtOAc were added and the organic layer was separated. The solvent was evaporated, and the crude material was purified via by reverse phase reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford the compound 6-isopropyl-10-methoxy-9-(3-methoxypropoxy)-6- methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylic acid (50 mg, 53 % yield) as a white solid. 1 H NMR (400 MHz, DMSO-de) δ 8.54 (s, 1 H), 7.52 (s, 1 H), 7.48 (s, 1 H), 7.06 (s, 1 H), 4.14 - 4.05 (m, 2H), 3.88 (s, 3H), 3.48 (t, J = 6.2 Hz, 2H), 3.25 (s, 3H), 3.18 (s, 2H), 1 .98 (dd, J = 12.6, 6.3 Hz, 2H), 1 .83 (dt, J = 13.6, 6.8 Hz, 1 H), 1 .66 (s, 3H), 0.81 (d, J = 6.8 Hz, 3H), 0.62 (d, J = 6.8 Hz, 3H). LCMS (ESI) m/z: 416.4 (M + 1)+.
[00685] Example 38
[00686] (S)-6-lsopropyl-10-methoxy-9-(3-methoxypropoxy)-6-methyl-2-oxo-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
[00687] Example 39
[00688] (R)-6-lsopropyl-10-methoxy-9-(3-methoxypropoxy)-6-methyl-2-oxo-6,7- dihvdro-2H-pyr -alisoquinoline-3-carboxylic acid
Figure imgf000146_0002
[00689] Step 1 : (S)-6-isopropyl-10-methoxy-9-(3-methoxypropoxy)-6-methyl-2-oxo- 6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid and (R)-6-isopropyl-10- methoxy-9-(3-methoxypropoxy)-6-methyl-2-oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3- carboxylic acid [00690] Ethyl 6-isopropyl-l 0-methoxy-9-(3-methoxypropoxy)-6-methyl-2-oxo-6,7- dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (220 mg, 0.474 mmol) was subjected to Chiral column [ Column: CHIRALCEL OZ-H; Column Size: 0.46 cm I.D. x 15 cm L; Injection: 3.0 μΙ; Mobile phase: MeOH/CAN/DEA=80/20/0.1 (v/v/v); Flow rate: 1 .0 mL/min; Wave length: UV 254 nm; Temperature: 35 °C] to give two isomers as colorless solids. The two enantiomers (100 mg, 0.215 mmol) were hydrolyzed independently in THF (2 mL), Water (0.5 mL) with LiOH (25 mg, 1 .07 mmol) at room temperature for 30 minutes. The reaction mixture was acidified with 1 N HCI to pH4, and the mixture was concentrated. Water and EtOAc were added and the organic layer was separated. The solvent was concentrated, and the crude material was purified via reverse phase reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford the compound.
[00691] Example 38, stereochemistry confirmed via VCD analysis. (S)-6-isopropyl- 10-methoxy-9-(3-methoxypropoxy)-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 - a]isoquinoline-3-carboxylic acid (50 mg, 48 % yield) as a white solid. 1H NMR (400 MHz, DMSO-de) δ 8.54 (s, 1 H), 7.52 (s, 1 H), 7.49 (s, 1 H), 7.06 (s, 1 H), 4.10 (td, J = 9.5, 3.2 Hz, 2H), 3.88 (s, 3H), 3.48 (t, J = 6.2 Hz, 2H), 3.25 (s,3H), 3.19 (d, J = 3.0 Hz, 2H), 2.02 - 1 .94 (m, 2H), 1 .83 (m, 1 H), 1 .66 (s, 3H), 0.81 (d, J = 6.8 Hz, 3H), 0.62 (d, J = 6.8 Hz, 3H). LCMS (ESI) m/z: 416.2 (M + 1)+.
[00692] Example 39, stereochemistry confirmed via VCD analysis. (R)-6-isopropyl- 10-methoxy-9-(3-methoxypropoxy)-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 - a]isoquinoline-3-carboxylic acid (40 mg, 38 % yield) as a white solid. 1H NMR (400 MHz, DMSO-de) δ 8.54 (s, 1 H), 7.52 (s, 1 H), 7.49 (s, 1 H), 7.06 (s, 1 H), 4.10 (dd, J = 12.2, 6.3 Hz, 2H), 3.88 (s, 3H), 3.48 (t, J = 6.2 Hz, 2H), 3.25 (s, 3H), 3.19 (d, J = 2.8 Hz, 2H), 1 .99 (dd, J = 12.6, 6.3 Hz, 2H), 1 .83 (dt, J = 13.5, 6.6 Hz, 1 H), 1 .66 (s, 3H), 0.82 (t, J = 7.7 Hz, 3H), 0.62 (d, J = 6.8 Hz, 3H). LCMS (ESI) m/z: 416.2 (M + 1)+.
[00693] Example 40
[00694] 9'-(3-Methoxypropoxy)-10'-methyl-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000147_0001
[00695] Step 1 : 4-(Benzyloxy)-3-hvdroxybenzaldehyde
Figure imgf000148_0001
[00696] To a stirred solution of 3,4-dihydroxybenzaldehyde (30 g, 217 mmol) in DMF (350 mL) was added K2CO3 (45 g, 326 mmol) and benzyl bromide (37.1 g, 217 mmol). The resulting mixture was heated to 55°C overnight, cooled to room temperature, concentrated and crude 4-(benzyloxy)-3-hydroxybenzaldehyde was used in the next step as is. 1H NMR (400 MHz, CDCI3) δ 9.85 (s, 1 H), 7.46 (d, J = 1 .9 Hz, 1 H), 7.44 - 7.38 (m, 6H), 7.04 (d, J = 8.3 Hz, 1 H), 5.77 (s, 1 H), 5.21 (s, 2H). LCMS (ESI) m/z: 229.2 (M + 1)+.
[00697] Step 2: -(Benzyloxy)-3-(3-methoxypropoxy)benzaldehvde
Figure imgf000148_0002
[00698] To crude 4-(benzyloxy)-3-hydroxybenzaldehyde from previous step was added K2C03 (45 g, 326 mmol) and 1 -bromo-3-methoxypropane (33.2 g, 217 mmol). The resulting mixture was heated to 70°C for 4 hrs, cooled to room temperature, the reaction mixture was filtered and the filtrate was partitioned between EtOAc and H20. The organic layers were separated and the combined organic layer was washed with brine, dried over Na2S04 and concentrated. The crude product which was purified by flash chromatography (silica gel, 0-40% EtOAc in petroleum ether) to afford the title compound (25 g, 38% yield). LCMS (ESI) m/z: 299.2 (M-1 )".
[00699] Step 3: -(Benzyloxy)-3-(3-methoxypropoxy)methanol
Figure imgf000148_0003
[00700] To a 0°C cooled solution of 4-(benzyloxy)-3-(3- methoxypropoxy)benzaldehyde (25 g, 83.3 mmol) in MeOH (300 mL) was added NaBH4 (3.4 g, 89.9 mmol) in portions. After addition, the mixture was allowed to warm up to room temperature and stirred for 30 minutes. The reaction mixture was quenched with ice water and partitioned between EtOAc and sat'd NH4CI. The organic layer was separated and the aqueous layer was extracted twice with EtOAc, the combined organic phases were washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by flash chromatography (silica gel, 0-40% EtOAc in petroleum ether) to afford the title compound (25 g, 99% yield). LCMS (ESI) m/z calcd for Ci8H2203: 302.37. Found: 301 .10(M-1)-. [00701] Step 5: 1 -(Benzyloxy)-4-(bromomethyr)-2-(3-methoxypropoxy)benzene
Figure imgf000149_0001
[00702] To a 0°C, solution of (4-(benzyloxy)-3-(3- methoxypropoxy)phenyl)methanol (25 g, 82.8 mmol) in chloroform (300 mL) was added TMSBr (12 mL, 91 mmol) dropwise. The resulting mixture was allowed to warm up to room temperature and stirred for 4 hrs. The reaction was quenched with ice water and partitioned between DCM and water, the organic layer was separated, aqueous layer was extracted with DCM, the combined organic layer was washed with brine, dried over Na2S04, filtered and concentrated. The residue was purified by flash chromatography (silica gel, 0-30% EtOAc in petroleum ether) to afford the title compound (9.8 g, 32% yield). 1H NMR (400 MHz, CDCI3) δ 7.37 - 7.32 (m, 2H), 7.31 - 7.25 (m, 2H), 7.25 - 7.18 (m, 1 H), 6.88 (d, J = 1 .9 Hz, 1 H), 6.80 (dd, J = 8.2, 1 .9 Hz, 1 H), 6.75 (d, J = 8.2 Hz, 1 H), 5.03 (s, 2H), 4.38 (s, 2H), 4.05 (t, J = 6.3 Hz, 2H), 3.50 (t, J = 6.2 Hz, 2H), 3.26 (s, 3H), 2.01 (m, 2H).
[00703] Step 6: Ethyl 1 -(4-(benzyloxy)-3-(3-methoxypropoxy)benzyl)cvclobutane-1 - carboxylate
Figure imgf000149_0002
[00704] To a -75 °C solution of ethyl cyclobutanecarboxylate (4.1 g, 32.2 mmol) in anhydrous THF (60 mL) was added LDA (20.1 mL, 2 mol/L in THF/hexane) dropwise. After stirring at the same temperature for 1 hour another solution of 1 - (benzyloxy)-4-(bromomethyl)-2-(3-methoxypropoxy)benzene (9.8 g, 26.8 mmol) in anhydrous THF (30 mL) was added dropwise. The resulting mixture was stirred at -75°C for 30 minutes then warmed up to room temperature gradually. The reaction was partitioned between EtOAc and sat'd. NH4CI, extracted with EtOAc. The organic layers were dried over Na2S04, filtered and concentrated to give the crude product which was purified by flash chromatography (silica gel, 0-30% EtOAc in petroleum ether) to afford the title compound (4.8 g, 43% yield). LCMS (ESI) m/z: 413.5 (M + 1 )+.
[00705] Step 7: 1 -(4-(Benzyloxy)-3-(3-methoxypropoxy)benzyl)cyclobutane-1 - carboxylic acid
Figure imgf000149_0003
[00706] To a solution of ethyl 1 -(4-(benzyloxy)-3-(3- methoxypropoxy)benzyl)cyclobutane-1 -carboxylate (4.8 g, 1 1 .6 mmol) in MeOH (60 mL) was added a solution of NaOH (1 .86 g, 46.4 mmol). The resulting mixture was heated to 55°C overnight. The reaction was quenched with 1 N HCI to pH = 6 in an ice-water bath, extracted with EtOAc. The organic layers were dried over Na2S04, filtered and concentrated and the residue was purified by flash chromatography (silica gel, 0-40% EtOAc in petroleum ether) to afford the title compound (4.4 g, 99% yield). LCMS (ESI) m/z: 385.2 (M + 1)+.
[00707] Step 8: 1 -(4-(Benzyloxy)-3-(3-methoxypropoxy)benzyl)cvclobutane-1 - carbonyl azide
Figure imgf000150_0001
[00708] At 0 °C, to a solution of 1 -(4-(benzyloxy)-3-(3- methoxypropoxy)benzyl)cyclobutane-1 -carboxylic acid (4.4 g, 1 1 .4 mmol) in anhydrous THF (80 mL) was added TEA (3.16 ml_, 22.8 mmol), followed by a solution of ethyl
carbonochloridate (1 .86 g, 17.1 mmol) in anhydrous THF (10 mL) dropwise, after stirred at the same temperature for 30 minutes another solution of sodium azide (1 .1 1 g, 17.1 mmol) in water (10 mL) was added dropwise. The resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was partitioned between toluene and water. The organic layer was separated, aqueous layer was extracted with toluene, the combined organic layers were washed with brine, filtered and concentrated to 100 mL volume, which was used to the next step directly. LCMS (ESI) m/z: 432.2 (M+23)+.
[00709] Step 9:
Figure imgf000150_0002
[00710] The crude solution from previous step was heated to reflux for 1 hour. After being cooled to room temperature, 50 mL of 4N HCI was added and the solution and heated to 60°C for 30 minutes. The solvent was removed and the residue was dissolved in DCM and treated with sat'd NaHC03. The organic layer was separated and the aqueous layer was extracted with DCM. The combined organic layers were dried over Na2S04, filtered and concentrated. The residue was purified by flash chromatography (silica gel, 0- 70% EtOAc in petroleum ether) to afford the title compound (2 g, 52% yield). LCMS (ESI) m/z: 354.2(M-1 )-. [00711] Step 10: Λ/-(1 -(4-(benzyloxy)-3-(3- methoxypropoxy)benzyl)cvclobutyl)formamide
Figure imgf000151_0001
[00712] To a 0°C solution of 1 -(4-(benzyloxy)-3-(3- methoxypropoxy)benzyl)cyclobutan-1 -amine (2 g, 5.87 mmol) in anhydrous THF (20 mL) was added acetic formic anhydride (2.34 g, 26.61 mmol) in THF (5 mL) dropwise. The resulting mixture was stirred at the same temperature for 30 minutes before was partitioned between DCM and water, organic layer was separated, aqueous layer was extracted with brine, the combined organic layer was dried over Na2S04, filtered and concentrated to give the crude product which was purified by flash chromatography (silica gel, 0-40% EtOAc in petroleum ether) to afford the title compound (1 .89 g, 84% yield). LCMS (ESI) m/z: 384.3 (M + 1 )+.
[00713] Step 1 1 : 7'-(Benzyloxy)-6'-(3-methoxypropoxy)-4'H-spiro[cvclobutane-1 ,3'- isoquinolinel
Figure imgf000151_0002
[00714] To a 0°C, cooled solution of /V-(1 -(4-(benzyloxy)-3-(3- methoxypropoxy)benzyl)cyclobutyl)formamide (1 .89 g, 4.93 mmol) in acetonitrile (20 mL) was added POCI3 (831 mg, 5.42 mmol) dropwise. The resulting mixture was heated to 90°C for 2 hours. The reaction mixture was concentrated, the residue was dissolved in DCM and washed with water. The organic layer was dried over Na2S04, filtered and concentrated to give 1 .43 g of the crude title compound which was used in the next step as is . LCMS (ESI) m/z: 366.4 (M + 1)+.
[00715] Step 12: Ethyl 10l-(benzyloxy)-9l-(3-methoxypropoxy)-2'-oxo-1 l.2l.7l.1 1 bl- tetrahydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000151_0003
[00716] To a solution of crude 7'-(benzyloxy)-6'-(3-methoxypropoxy)-4'/-/- spiro[cyclobutane-1 ,3'-isoquinoline] (1 .43 g, 3.91 mmol) in ethanol (30 mL) was added ethyl 2-(ethoxymethylene)-3-oxobutanoate (2.54 g, 13.69 mmol). The resulting mixture was heated to 84°C overnight. After removal of the solvent, the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (1 .73 g, 88% yield). LCMS (ESI) m/z:: 506.3 (M + 1 )+.
[00717] Step 13: Ethyl 10'-(benzyloxy)-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000152_0001
[00718] To a stirred solution of ethyl 10'-(benzyloxy)-9'-(3-methoxypropoxy)-
2'-oxo-1 ',2',7',1 1 b'-tetrahydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (1 .73 g, 3.42 mmol) in DME (30 mL) was added p-chloranil (840 mg, 3.42 mmol). The resulting mixture was heated to 100°C overnight. After removal of the solvent, the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (1 .74 g, 100% yield). LCMS (ESI) m/z: 504.4 (M + 1)+.
[00719] Step14: Ethyl 10'-hvdroxy-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000152_0002
[00720] To a solution of ethyl 10'-(benzyloxy)-9'-(3-methoxypropoxy)-2'-oxo-
2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (1 .74 g, 3.42 mmol) in MeOH (30 mL) was added palladium (100 mg, 5% on carbon (wet with ca. 55% Water)), the resulting mixture was stirred at room temperature overnight. The Palladium was removed by filtering, the filtrate was concentrated and the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (1 .4 g, 99% yield). LCMS (ESI) m/z: 414.2 (M + 1 )+.
[00721] Step 15: Ethyl 9'-(3-methoxypropoxy)-2'-oxo-10'- (((trifluoromethyl')sulfonyl')oxy')-2',7'-dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel- 3'-carboxylate
Figure imgf000153_0001
[00722] To a stirred mixture of ethyl 10'-hydroxy-9'-(3-methoxypropoxy)-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (1 .4 g, 3.455 mmol) and Cs2C03 (2.27g, 6.91 mmol) in anhydrous DMF (20 mL),PhNTf2 (1 .48 g, 4.146 mmol) was added at 0 °C. The resulting mixture was stirred at room temperature for 1 h. Then the mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and extracted with EtOAc (50ml_x2). The combined organic solution was washed with brine, dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel, 0-10% MeOH in DCM) to afford ethyl 9'-(3-methoxypropoxy)-2'-oxo-10'- (((trifluoromethyl)sulfonyl)oxy)-2',7'- dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (886 mg, 49% yield) as a yellow solid. LCMS (ESI) m/z: 546.4 (M + 1 )+.
[00723] Step 16: Ethyl 9'-(3-methoxypropoxy)-10'-methyl-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000153_0002
[00724] To a solution of ethyl 9'-(3-methoxypropoxy)-2'-oxo-10'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate (120 mg, 0.220 mmol) in dioxane (5 ml), methylboronic acid (129.8 mg, 2.2mmol), Pd(dppf)CI2 (16.1 mg,0.022mmol), K3P04 (93.4 mg,0.440mmol), H20(1 drop) were added. The resulting mixture was stirred at 100 oC for 8 h. The mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50ml_x2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography(silica gel,0~10%MeOH in DCM) to afford ethyl 9'- (3-methoxypropoxy)-10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (60 mg, 66.3% yield) as a yellow oil. LCMS (ESI) m/z: 412.6 (M + 1 )+. [00725] Step 17: 9'-(3-Methoxypropoxy)-10'-methyl-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000154_0001
[00726] To a solution of ethyl 9'-(3-methoxypropoxy)-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate
[00727] (60 mg, 0.146 mmol) in MeOH(5 ml), NaOH (29.1 mg, 0.729 mmol) dissolved in H20(1 ml) was added. The resulting mixture was stirred at room temperature for 2 h. The reaction mixture was acidified to ph~5 with 1 N HCI. Then the mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50ml_x2). The combined organic solution was dried over anhydrous Na2S04 and concentrated. The residue was purified by reverse phase reverse phase HPLC to give (C18
column,40%~100%MeCN in H20,with 0.1 % formic acid in H20) to give 9'-(3- methoxypropoxy)-10'-methyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid (18.4 mg, 32.9% yield) as a white solid. LCMS (ESI) m/z: 384.3 (M + 1)+. Ή NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1 H), 7.86 (s, 1 H), 7.25 (s, 1 H), 7.04 (s, 1 H), 4.14 (t, J = 6.2 Hz, 2H), 3.51 (t, J = 6.1 Hz, 2H), 3.32 (s, 2H), 3.26 (s, 3H), 2.63 (dd, J = 19.8, 9.9 Hz, 2H), 2.20 (s, 3H), 2.05 - 1 .98 (m, 4H), 1 .93 - 1 .85 (m, 2H).
[00728] Example 41
[00729] 10'-Ethvnyl-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000154_0002
[00730] Step 1 : Ethyl 9'-(3-methoxypropoxyV2'-oxo-10'-((trimethylsilyl)ethvnylV2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000155_0001
[00731] A mixture of ethyl 9'-(3-methoxypropoxy)-2'-oxo-10'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate (100 mg, 0.18 mmol) in DMF (1 mL), trimethylsilylacetylene (24 mg, 0.24 mmol) and Cul (10 mg), PdCI2(PPh3)2 (10 mg) was stirred at 60°C for 6h. The resulting mixture was partitioned between EtOAc and H20. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by flash chromatography (silica gel, 0-50% DCM in MeOH) to afford the title compound (40 mg, 45 % yield). LCMS (ESI) m/z: 494.5 (M + 1 )+.
[00732] Step 2: 10'-Ethvnyl-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000155_0002
[00733] To a solution of ethyl 9'-(3-methoxypropoxy)-2'-oxo-10'-
((trimethylsilyl)ethynyl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (19 mg, 0.04 mmol) in THF (2 mL) was added LiOH (4 mg, 0.15 mmol) and H20 (0.5 ml). After stirred at room temperature for 2 h, the reaction mixture was neutralized to pH = 7. The solvents were removed and the residue was purified by reverse phase HPLC (C18, 40-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (9.3 mg, 62% yield) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1 H),8.10 (s, 1 H), 7.34 (s, 1 H), 7.21 (s, 1 H), 4.34 (s, 1 H), 4.21 -4.18 (m, 2H), 3.56-3.47 (m, 2H), 3.38 (s, 2H) 3.26 (s, 3H), 2.69-2.61 (m, 2H), 2.09-1 .95 (m, 4H), 1 .94-1 .82 (m, 2H) .LCMS (ESI) m/z: 394.2 (M + 1)+.
[00734] Example 42 [00735] 9'-(3-MethoxypropoxyV2'-oxo-2',7'-dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -
Figure imgf000156_0001
[00736] Step 1 : 4-(Benzyloxy)-3-hvdroxybenzaldehyde
Figure imgf000156_0002
[00737] To a stirred solution of 3,4-dihydroxybenzaldehyde (30 g, 217 mmol) in DMF (350 mL) was added K2C03 (45 g, 326 mmol) and benzyl bromide (37.1 g, 217 mmol). The resulting mixture was heated to 55°C overnight, cooled to room temperature, concentrated and crude 4-(benzyloxy)-3-hydroxybenzaldehyde was used in the next step as is. Ή NMR (400 MHz, CDCI3) δ 9.85 (s, 1 H), 7.46 (d, J = 1 .9 Hz, 1 H), 7.44 - 7.38 (m, 6H), 7.04 (d, J = 8.3 Hz, 1 H), 5.77 (s, 1 H), 5.21 (s, 2H).. LCMS (ESI) m/z: 229.2 (M + 1)+.
[00738] Step 2: -(Benzyloxy)-3-(3-methoxypropoxy)benzaldehyde
Figure imgf000156_0003
[00739] To crude 4-(benzyloxy)-3-hydroxybenzaldehyde from previous step was added K2C03 (45 g, 326 mmol) and 1 -bromo-3-methoxypropane (33.2 g, 217 mmol). The resulting mixture was heated to 70°C for 4 hrs, cooled to room temperature, the reaction mixture was filtered and the filtrate was partitioned between EtOAc and H2O. The organic layers were separated and the combined organic layer was washed with brine, dried over Na2S04 and concentrated. The crude product which was purified by flash chromatography (silica gel, 0-40% EtOAc in petroleum ether) to afford the title compound (25 g, 38% yield). LCMS (ESI) m/z: 299.2 (M-1 )".
[00740] Step 3: -(Benzyloxy)-3-(3-methoxypropoxy)methanol
Figure imgf000156_0004
[00741] To a 0°C cooled solution of 4-(benzyloxy)-3-(3- methoxypropoxy)benzaldehyde (25 g, 83.3 mmol) in MeOH (300 mL) was added NaBH4 (3.4 g, 89.9 mmol) in portions. After addition, the mixture was allowed to warm up to room temperature and stirred for 30 minutes. The reaction mixture was quenched with ice water and partitioned between EtOAc and sat'd NH4CI. The organic layer was separated and the aqueous layer was extracted twice with EtOAc, the combined organic phases were washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by flash chromatography (silica gel, 0-40% EtOAc in petroleum ether) to afford the title compound (25 g, 99% yield). LCMS (ESI) m/z: 301 .1 (M-1)".
[00742] Step 5: -(Benzyloxy)-4-(bromomethyl)-2-(3-methoxypropoxy)benzene
Figure imgf000157_0001
[00743] To a 0°C, solution of (4-(benzyloxy)-3-(3- methoxypropoxy)phenyl)methanol (25 g, 82.8 mmol) in chloroform (300 mL) was added TMSBr (12 mL, 91 mmol) dropwise. The resulting mixture was allowed to warm up to room temperature and stirred for 4 hrs. The reaction was quenched with ice water and partitioned between DCM and water, the organic layer was separated, aqueous layer was extracted with DCM, the combined organic layer was washed with brine, dried over Na2S04, filtered and concentrated. The residue was purified by flash chromatography (silica gel, 0-30% EtOAc in petroleum ether) to afford the title compound (9.8 g, 32% yield). 1H NMR (400 MHz, CDCb) δ 7.37 - 7.32 (m, 2H), 7.31 - 7.25 (m, 2H), 7.25 - 7.18 (m, 1 H), 6.88 (d, J = 1 .9 Hz, 1 H), 6.80 (dd, J = 8.2, 1 .9 Hz, 1 H), 6.75 (d, J = 8.2 Hz, 1 H), 5.03 (s, 2H), 4.38 (s, 2H), 4.05 (t, J = 6.3 Hz, 2H), 3.50 (t, J = 6.2 Hz, 2H), 3.26 (s, 3H), 2.01 (m, 2H).
[00744] Step 6: Ethyl 1 -(4-(benzyloxy)-3-(3-methoxypropoxy)benzyl)cvclobutane-1 - carboxylate
Figure imgf000157_0002
[00745] To a -75°C solution of ethyl cyclobutanecarboxylate (4.1 g, 32.2 mmol) in anhydrous THF (60 mL) was added LDA (20.1 mL, 2 mol/L in THF/hexane) dropwise. After stirring at the same temperature for 1 hour another solution of 1 - (benzyloxy)-4-(bromomethyl)-2-(3-methoxypropoxy)benzene (9.8 g, 26.8 mmol) in anhydrous THF (30 mL) was added dropwise. The resulting mixture was stirred at -75°C for 30 minutes then warmed up to room temperature gradually. The reaction was partitioned between EtOAc and sat'd. NH4CI, extracted with EtOAc. The organic layers were dried over Na2S04, filtered and concentrated to give the crude product which was purified by flash chromatography (silica gel, 0-30% EtOAc in petroleum ether) to afford the title compound (4.8 g, 43% yield). LCMS (ESI) m/z: 413.5 (M + 1 )+. [00746] Step 7: 1 -(4-(Benzyloxy)-3-(3-methoxypropoxy)benzvDcvclobutane-1■ carboxylic acid
Figure imgf000158_0001
[00747] To a solution of ethyl 1 -(4-(benzyloxy)-3-(3- methoxypropoxy)benzyl)cyclobutane-1 -carboxylate (4.8 g, 1 1 .6 mmol) in MeOH (60 ml_) was added a solution of NaOH (1 .86 g, 46.4 mmol). The resulting mixture was heated to 55°C overnight. The reaction was quenched with 1 N HCI to pH 6 in an ice-water bath, extracted with EtOAc. The organic layers were dried over Na2S04, filtered and concentrated and the residue was purified by flash chromatography (silica gel, 0-40% EtOAc in petroleum ether) to afford the title compound (4.4 g, 99% yield). LCMS (ESI) m/z: 385.2 (M + 1)+.
[00748] Step 8: 1 -(4-(Benzyloxy)-3-(3-methoxypropoxy)benzyl)cyclobutane-1 - carbonyl azide
Figure imgf000158_0002
[00749] At 0°C, to a solution of 1 -(4-(benzyloxy)-3-(3- methoxypropoxy)benzyl)cyclobutane-1 -carboxylic acid (4.4 g, 1 1 .4 mmol) in anhydrous THF (80 mL) was added TEA (3.16 ml_, 22.8 mmol), followed by a solution of ethyl
carbonochloridate (1 .86 g, 17.1 mmol) in anhydrous THF (10 mL) dropwise, after stirred at the same temperature for 30 minutes another solution of sodium azide (1 .1 1 g, 17.1 mmol) in water (10 mL) was added dropwise. The resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was partitioned between toluene and water. The organic layer was separated, aqueous layer was extracted with toluene, the combined organic layers were washed with brine, filtered and concentrated to 100 mL volume, which was used to the next step directly. LCMS (ESI) m/z: 432.2 (M+23)+.
[00750] Step 9: 1 -(4-(Benzyloxy)-3-(3-methoxypropoxy)benzyl)cvclobutan-1 -amine
Figure imgf000158_0003
[00751] The crude solution from previous step was heated to reflux for 1 hour. After being cooled to room temperature, 50 mL of 4N HCI was added and the solution and heated to 60°C for 30 minutes. The solvent was removed and the residue was dissolved in DCM and treated with sat'd NaHC03. The organic layer was separated and the aqueous layer was extracted with DCM. The combined organic layers were dried over Na2S04, filtered and concentrated. The residue was purified by flash chromatography (silica gel, 0- 70% EtOAc in petroleum ether) to afford the title compound (2 g, 52% yield). LCMS (ESI) m/z: 354.2 (M-1)".
[00752] Step 10: Λ/-(1 -(4-(benzyloxy)-3-(3- methoxypropoxy)benzyl)cvclobutyl)formamide
Figure imgf000159_0001
[00753] To a 0°C solution of 1 -(4-(benzyloxy)-3-(3- methoxypropoxy)benzyl)cyclobutan-1 -amine (2 g, 5.87 mmol) in anhydrous THF (20 mL) was added acetic formic anhydride (2.34 g, 26.61 mmol) in THF (5 mL) dropwise. The resulting mixture was stirred at the same temperature for 30 minutes before was partitioned between DCM and water, organic layer was separated, aqueous layer was extracted with brine, the combined organic layer was dried over Na2S04, filtered and concentrated to give the crude product which was purified by flash chromatography (silica gel, 0-40% EtOAc in petroleum ether) to afford the title compound (1 .89 g, 84% yield). LCMS (ESI) m/z: 384.3 (M + 1 )+.
[00754] Step 1 1 : 7'-(Benzyloxy)-6'-(3-methoxypropoxy)-4'H-spiro[cvclobutane-1 ,3'- isoquinolinel
Figure imgf000159_0002
[00755] To a 0°C, cooled solution of /V-(1 -(4-(benzyloxy)-3-(3- methoxypropoxy)benzyl)cyclobutyl)formamide (1 .89 g, 4.93 mmol) in acetonitrile (20 mL) was added POCI3 (831 mg, 5.42 mmol) dropwise. The resulting mixture was heated to 90°C for 2 hours. The reaction mixture was concentrated, the residue was dissolved in DCM and washed with water. The organic layer was dried over Na2S04, filtered and concentrated to give 1 .43 g of the crude title compound which was used in the next step as is. LCMS (ESI) m/z: 366.4 (M + 1)+.
[00756] Step 12: Ethyl 10l-(benzyloxy)-9l-(3-methoxypropoxy)-2'-oxo-1 l.2l.7l.1 1 bl- tetrahvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000159_0003
[00757] To a solution of crude 7'-(benzyloxy)-6'-(3-methoxypropoxy)-4'/-/- spiro[cyclobutane-1 ,3'-isoquinoline] (1 .43 g, 3.91 mmol) in ethanol (30 mL) was added ethyl 2-(ethoxymethylene)-3-oxobutanoate (2.54 g, 13.69 mmol). The resulting mixture was heated to 84°C overnight. After removal of the solvent, the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (1 .73 g, 88% yield). LCMS (ESI) m/z: 506.3 (M + 1 )+.
[00758] Step 13: Ethyl 10'-(benzyloxy)-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000160_0001
[00759] To a stirred solution of ethyl 10'-(benzyloxy)-9'-(3-methoxypropoxy)-
2'-oxo-1 ',2',7',1 1 b'-tetrahydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (1 .73 g, 3.42 mmol) in DME (30 mL) was added p-chloranil (840 mg, 3.42 mmol). The resulting mixture was heated to 100°C overnight. After removal of the solvent, the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (1 .74 g, 100% yield). LCMS (ESI) m/z: 504.4 (M + 1)+.
[00760] Step 14: Ethyl 10'-hvdroxy-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000160_0002
[00761] To a solution of ethyl 10'-(benzyloxy)-9'-(3-methoxypropoxy)-2'-oxo-
2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (1 .74 g, 3.42 mmol) in MeOH (30 mL) was added palladium (100 mg, 5% on carbon (wet with ca. 55% Water)), the resulting mixture was stirred at room temperature overnight. The Palladium was removed by filtering, the filtrate was concentrated and the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (1 .4 g, 99% yield). LCMS (ESI) m/z: 414.2 (M + 1 )+. [00762] Step 15: Ethyl 9'-(3-methoxypropoxy)-2'-oxo-10'- (((trifluoromethyl')sulfonyl')oxy')-2',7'-dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel- 3'-carboxylate
Figure imgf000161_0001
[00763] To a stirred mixture of ethyl 10'-hydroxy-9'-(3-methoxypropoxy)-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (1 .4 g, 3.455 mmol) and Cs2C03 (2.27g, 6.91 mmol) in anhydrous DMF (20 ml_), PhNTf2 (1 .48 g, 4.146 mmol) was added at 0 °C. The resulting mixture was stirred at room temperature for 1 h. Then the mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and extracted with EtOAc (50ml_x2). The combined organic solution was washed with brine, dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel , 0-10% MeOH in DCM) to afford ethyl 9'-(3-methoxypropoxy)-2'-oxo-10'- (((trifluoromethyl)sulfonyl)oxy)-2',7'- dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (886 mg, 49% yield) as a yellow solid. LCMS (ESI) m/z: 546.4 (M + 1 )+.
[00764] Step 16: Ethyl 9'-(3-methoxypropoxy)-2'-oxo-10'-vinyl-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000161_0002
[00765] To a solution of ethyl 9'-(3-methoxypropoxy)-2'-oxo-10'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate (886 mg, 1 .62 mmol) in dioxane (5 ml_) was added 4,4,5,5-tetramethyl-2- vinyl-1 ,3,2-dioxaborolane (375 mg, 2.44 mmol), KOAc (318 mg, 3.24 mmol),
Pd(dppf)CI2 DCM (130 mg, 0.16 mmol) and 1 drop of water. The resulting mixture was heated to 85°C for 2 hours. After cooling to room temperature, the solvent was removed and the residue was purified by flash chromatography (silica gel, 0-30% DCM in MeOH) to afford the title compound (630 mg, 92% yield). LCMS (ESI) m/z: 424.7 (M + 1 )+. [00766] Step 17: Ethyl 10'-formyl-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000162_0001
[00767] To a 0 °C solution of ethyl 9'-(3-methoxypropoxy)-2'-oxo-10'-vinyl-
2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (423 mg, 1 mmol) in THF (15 ml_) and water (4 ml_) was added K20s04 «2H20 (35 mg, 0.1 mmol). After stirred for 2 minutes, Nal04 (642 mg, 3 mmol) was added portions. The reaction was quenched with sat'd Na2S03 after stirred at room temperature for 2 hours. The reaction mixture was extracted twice with DCM, the combined organic layers were dried over Na2S04, and was concentrated. The crude material was purified by flash chromatography (silica gel, 0-30% MeOH in DCM) to afford the title compound (400 mg, 94% yield). LCMS (ESI) m/z: 426.5 (M + 1)+.
[00768] Step 18: 3'-(Ethoxycarbonyl)-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-10'-carboxylic acid
Figure imgf000162_0002
[00769] To a 0°C solution of ethyl 10'-formyl-9'-(3-methoxypropoxy)-2'-oxo-
2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.26 mmol) in THF (2 ml_) was added a solution of NaCI02 (128 mg, 1 .41 mmol), NaH2P04 (169 mg, 1 .41 mmol) in water (2 ml_), followed by a solution of 3-methyl-1 -butene (0.5 ml_) in tBuOH (0.5 ml_) and THF (2 ml_). After stirring at the same temperature for 15 minutes, the reaction was partitioned between ethyl acatate and H20. The organic layer was separated and the aqueous layer was extracted with EA. The combined organic layers were dried over Na2S04, filtered and concentrated to give the crude product which was purified by flash chromatography (silica gel, 0-30% MeOH in DCM) to afford the title compound (100 mg, 88% yield). LCMS (ESI) m/z: 442.5 (M + 1)+. [00770] Step 19: 9'-(3-Methoxypropoxy)-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolinel-3',10'-dicarboxylic acid
Figure imgf000163_0001
[00771] To a solution of 3'-(ethoxycarbonyl)-9'-(3-methoxypropoxy)-2'-oxo-
2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-10'-carboxylic acid (70 mg, 0.16 mmol) in MeOH (4 mL) was added NaOH(25 mg, 0.64 mmol) and H20 (1 ml). After stirring at room temperature for 30 minutes, the reaction mixture was treated with 1 N HCI to achieve pH = 7. The solvent was removed and the residue was purified by reverse phase reverse phase HPLC (Gilson, C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (33 mg, 50% yield) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 8.80 (s, 1 H), 8.14 (s, 1 H), 7.24 (s, 2H), 4.19 (t, J=6.0 Hz, 2H), 3.52 (t, J=6.1 Hz, 2H), 3.40 (s, 2H), 3.25 (s, 3H), 2.69-2.63 (m, 2H), 2.05-1 .97 (m , 4H ), 1.93-1.86 (m, 2H). LCMS (ESI) m/z: 414.3 (M + 1)+.
[00772] Example 43
[00773] 9'-(3-Methoxypropoxy)-10'-(methylcarbamoyl)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000163_0002
[00774] Step 1 : Ethyl 9'-(3-methoxypropoxy)-10'-(methylcarbamoyl)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000163_0003
[00775] To a solution of 3'-(ethoxycarbonyl)-9'-(3-methoxypropoxy)-2'-oxo-
2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-10'-carboxylic acid (150 mg, 0.34 mmol) in DCM (2 mL) was added HBTU (258 mg, 0.68 mmol), followed by methanamine (0.34 mL, 2 mol/L in THF). After stirring at room temperature for 15 minutes, the reaction was partitioned between DCM and H20. The organic layer was separated and the aqueous layer was extracted with DCM. The combined organic layers were dried over Na2S04, concentrated and the crude product was purified by flash chromatography (silica gel, 0-10% MeOH in DCM) to afford the title compound (150 mg, 97% yield). LCMS (ESI) m/z: 455.5 (M + 1)+.
[00776] Step 2: 9'-(3-Methoxypropoxy)-10'-(methylcarbamoyl)-2'-oxo-2',7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000164_0001
[00777] To a solution of ethyl 9'-(3-methoxypropoxy)-10'-(methylcarbamoyl)-
2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (150 mg, 0.33 mmol) in MeOH (10 mL) was added NaOH (53 mg, 1.32 mmol) and H20 (2 ml). After stirring at room temperature for 30 minutes, the reaction mixture was acidified with 1 N HCI to pH = 5. The solvent was removed and the residue was purified by reverse phase reverse phase HPLC (Gilson, C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (50 mg, 35% yield) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 8.81 (s, 1 H), 8.22-8.15 (m, 2H), 726 (s, 1 H), 7.20 (s, 1 H), 4.25 (t, J=6.1 Hz, 2H), 3.52 (t, J=5.9 Hz, 2H), 3.40 (s, 2H), 3.28 (s, 3H), 2.82 (d, J=4.7 Hz, 3H), 2.70-2.62 (m, 2H), 2.07-1.98 (m , 4H ), 1.93-1 .86 (m, 2H). LCMS (ESI) m/z: 427.3 (M + 1)+.
[00778] Example 44
[00779] 10'-Carbamoyl-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000164_0002
[00780] Step 1 : Ethyl 10'-carbamoyl-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000164_0003
[00781] To a solution of 3'-(ethoxycarbonyl)-9'-(3-methoxypropoxy)-2'-oxo-
2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-10'-carboxylic acid (60 mg, 0.136 mmol) in DMF (4 mL) was added NH4CI (36 mg, 0.68 mmol), EDCI(39 mg, 0.2 mmol), HOBT (27 mg, 0.2 mmol) and DIPEA (141 mg, 1 .088 mmol). The resulting mixture was stirred at room temperature overnight. After partitioning between DCM and H20, the reaction was extracted with 15% IPA in DCM, the combined organic layers were dried over Na2S04, filtered and concentrated to give the crude material which was purified by flash chromatography (silica gel, 0-40% MeOH in DCM) to afford the title compound (16 mg, 27% yield). LCMS (ESI) m/z: 441 .5 (M + 1 )+.
[00782] Step 2: 10'-Carbamoyl-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospirofcyclobuta '-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000165_0001
[00783] To a solution of ethyl 10'-carbamoyl-9'-(3-methoxypropoxy)-2'-oxo-
2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (16 mg, 0.038mmol) in THF (4 mL) was added LiOH (4 mg, 0.15 mmol) and H20 (1 ml). After stirred at room temperature for 30 minutes, the reaction mixture was acidified with 1 N HCI to pH=5. The solvent was removed and the residue was purified by reverse phase reverse phase HPLC (Gilson, C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (10 mg, 64% yield) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 8.83 - 8.79 (m, 1 H), 8.26 (s, 1 H), 7.76 (s, 1 H), 7.66 (s, 1 H), 7.27 (s, 1 H), 7.18 (s, 1 H), 4.27 (t, J = 6.1 Hz, 2H), 3.52 (t, J = 5.9 Hz, 2H), 3.41 (s, 2H), 3.26 (s, 3H), 2.67 (dd, J = 8.0, 2.4 Hz, 2H), 2.08 - 1 .99 (m, 4H), 1 .94 - 1 .87 (m, 2H). LCMS (ESI) m/z: 413.4 (M + 1 )+.
[00784] Example 45
[00785] 10'-Acetyl-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolin -3'-carboxylic acid
Figure imgf000165_0002
[00786] Step 1 : Ethyl 10'-acetyl-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000166_0001
[00787] To a solution of ethyl 9'-(3-methoxypropoxy)-2'-oxo-10'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate (300 mg, 0.55mmol) in DMF (6 mL) was added l -(vinyloxy) butane (1 10 mg, 1 .1 mmol), 1 ,3-bis(diphenylphosphino)propane (27 mg, 0.066 mmol), Pd(OAc)2 (12 mg, 0.06 mmol) and TEA (1 1 1 mg, 1 .1 mmol). The resulting mixture was heated at 90 °C overnight. After cooling to room temperature, the reaction was treated with 1 N HCI and extracted with EA. The combined organic layers were washed with brine, dried over Na2S04, filtered and concentrated. The residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM to afford the title compound. (206 mg, 85% yield). LCMS (ESI) m/z: 440.3 (M + 1)+.
[00788] Step 2: 10'-Acetyl-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000166_0002
[00789] To a solution of ethyl 10'-acetyl-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (70 mg, 0.16 mmol) in THF (7 mL) was added LiOH (15 mg, 0.6 4mmol) in water (1 mL), The resulting mixture was stirred at room temperature for 20 minutes. The reaction mixture was acidified with 1 N HCI to pH 6, extracted with 15% IPA in DCM twice, the combined organic layers were dried over Na2S04, filtered and concentrated. The residue was purified by reverse phase reverse phase HPLC (Gilson, C18, 0%~100% MeCN in water with 0.1 % formic acid) to afford the title compound. (35 mg, 54% yield) as a white solid. LCMS (ESI) m/z: 412.4 (M + 1 )+. 1H NMR (400 MHz, DMSO-d6) δ 8.81 (s, 1 H), 8.12 (s, 1 H), 7.32 (s, 1 H), 7.26 (s, 1 H), 4.26 (t, J = 6.2 Hz, 2H), 3.52 (t, J = 6.1 Hz, 2H), 3.42 (s, 2H), 3.27 (s, 3H), 2.70 - 2.63 (m, 2H), 2.58 (s, 3H), 2.08 - 1 .99 (m, 4H), 1 .94 - 1 .86 (m, 2H). [00790] Example 46
[00791] 3-lsopropoxy-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000167_0001
[00792] Step 1 : Ethyl 3-isopropoxy-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000167_0002
[00793] To a solution of ethyl 3-hydroxy-10'-methoxy-9'-(3-methoxypropoxy)-
2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (120 mg, 0.27 mmol) in 2-iodopropane (919 mg, 5.4 mmol) was added Ag20 (125 mg, 0.54 mmol). The resulting mixture was stirred at room temperature for 4 days before it was diluted with diethyl ether, then filtered off. The filtrate was concentrated and the residue was purified by flash chromatography (silica gel, 0~20% MeOH in DCM) to afford the title compound. (65 mg, 50%), LCMS (ESI) m/z: 486.4 (M + 1)+.
[00794] Step 2: 3-isopropoxy-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000167_0003
[00795] To a solution of ethyl 3-isopropoxy-10'-methoxy-9'-(3-methoxypropoxy)-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (65 mg, 0.13 mmol) in MeOH (4 mL) was added NaOH (20 mg, 0.52 mmol) in water (1 ml_). The resulting mixture was stirred at room temperature for 30 minutes, and then was quenched with 1 N HCI to pH 5. The solvent was removed and the residue was purified by reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (40 mg, 68% yield) as an off-white solid. 8.22 (U210042-047-1) 1H NMR (400 MHz, DMSO-de) δ 8.74 (s, 1 H), 7.49 (s, 1 H), 7.44 (s, 1 H), 7.01 (s, 1 H), 4.27 - 4.05 (m, 1 H), 4.1 1 (t, J = 6.4 Hz, 2H), 3.88 (s, 3H), 3.62 - 3.56 (m, 1 H), 3.48 (t, J = 6.2 Hz, 2H), 3.29-3.17 (s, 2H), 3.25 (s, 3H), 2.91 - 2.65 (m, 2H), 2.43 - 2.16 (m, 2H), 2.02 - 1 .97 (m, 2H), 1 .08 (dd, J = 6.1 , 3.5 Hz, 6H). LCMS (ESI) m/z: 458.7 (M + 1)+.
[00796] Example 47
[00797] 3.10'-Dimethoxy-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000168_0001
[00798] Step 1 : 3-Ethoxy-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000168_0002
[00799] To a solution of ethyl 3-hydroxy-10'-methoxy-9'-(3-methoxypropoxy)-
2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (1 1 1 mg, 0.25 mmol) in DMF (1 .0 mL) was added NaH (60% mixture with mineral oil, 25 mg, 0.625 mmol). The resulting mixture was stirred at rt for 30 min before Etl (24 mg, 0.3 mmol) was added. After stirred at room temperature for 3 h, the reaction mixture was diluted with MeOH/H20 (0.5 mL / 0.5 mL) and continued with stirring for 2 h. The reaction mixture was acidified with 1 N HCI, and the mixture was extracted with CH2CI2 (3x5 mL). The combined organic layers were washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase reverse phase HPLC (C18, 0-90% MeCN in H20 with 0.1 % formic acid) to afford the title compound (41 .9 mg, 37% yield over two steps) as a white powder. LCMS (ESI) m/z: 444.3 (M + 1)+. Ή NMR (400 MHz, CDCI3) δ 16.06 - 15.92 (s, 1 H), 8.92 - 8.83 (s, 1 H), 7.17 - 7.12 (s, 1 H), 7.08 - 7.02 (s, 1 H), 6.83 - 6.76 (s, 1 H), 4.28 - 3.94 (m, 1 H), 4.22 - 4.18 (m, 2H), 3.94 -3.89 (m, 3H), 3.59 (t, J = 6.0 Hz, 2H), 3.43 (q, J = 6.8 Hz, 2H), 3.37 (s, 3H), 3.27 -3.04 (s, 2H), 2.89 - 2.78 (m, 1 H), 2.60 - 2.50 (m, 1 H), 2.31 (dd, J = 14.4, 4.8 Hz, 2H), 2.15 (p, J = 6.0 Hz, 3H), 1 .27 - 1 .19 (m, 3H). [00800] Example 48
[00801] 3-(Cvclopentyloxy)-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000169_0001
[00802] Step 1 : 3-(Cvclopentyloxy)-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000169_0002
[00803] To a mixture of ethyl 3-hydroxy-10'-methoxy-9'-(3-methoxypropoxy)-
2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (1 1 1 mg, 0.25 mmol) and bromocyclopentane (1 .47 g, 9.88 mmol) in DMF (1 .0 mL) was added Ag20 (0.058 g, 0.25 mmol). The resulting mixture was stirred at 80 °C in the dark for 24 h before it was concentrated under reduced pressure. The brown yellow viscous oil residue was dissolved in MeOH/H20 (1 .0 mL/1 .0 mL), followed by addition of NaOH (20 mg, 0.5 mmol). The resulting mixture was stirred at room temperature for 4 h, and then filtered with a Biichner funnel. The collected filtrate was concentrated and acidified with 1 N HCI, and the obtained mixture was extracted with CH2CI2 (3x5 mL). The combined organic layers were washed with brine, dried over Na2S04, concentrated and the crude product was purified by reverse phase reverse phase HPLC (C18, 0-90% MeCN in H20 with 0.1 % formic acid) to afford the title compound (5.0 mg, 4% yield over two steps) as an off-white solid. LCMS (ESI) m/z: 484.3 (M + 1)+. Ή NMR (400 MHz, DMSO-d6) δ 8.75 - 8.73 (s, 1 H), 8.14 - 6.53 (s, 1 H), 7.49 (s, 1 H), 7.45 - 7.42 (s, 1 H), 7.02 (s, 1 H), 4.1 1 (t, J = 6.0 Hz, 2H), 4.06 - 3.99 (m, 1 H), 3.94 - 3.83 (m, 1 H), 3.88 (s, 3H), 3.48 (t, J = 6.0 Hz, 2H), 3.26 (s, 3H), 3.17 (s, 2H), 2.86 (dd, J = 13.2, 6.4 Hz, 1 H), 2.46 - 2.39 (m, 2H), 2.17 (dd, J = 13.6, 5.6 Hz, 1 H), 2.04 - 1 .96 (m, 2H), 1 .67 - 1 .43 (m, 8H).
[00804] Example 49 [00805] 10'-lsopropoxy-9'-(3-methoxypropoxy')-2'-oxo-2',7'-dihvdrospiro[cvclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000170_0001
[00806] Step 1 : Ethyl 10'-isopropoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000170_0002
[00807] To a solution of ethyl 10'-hydroxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (90 mg, 0.22 mmol) in DMF (3 mL) was added K2C03 (91 mg, 0.66 mmol) and 2-bromopropane (41 mg, 0.33 mmol). The resulting mixture was heated to 90°C overnight. After cooling to room temperature, the reaction mixture was partitioned between DCM and sat. NH4CI. The organic layer was separated and the aqueous layer was extracted with DCM. The combined organic layers were dried with Na2S04, concentrated, and the residue was purified by flash chromatography (silica gel, 0~20% MeOH in DCM) to afford the title compound. (45 mg, 45% yield). LCMS (ESI) m/z: 456.4 (M + 1)+.
[00808] Step 2: 10'-lsopropoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000170_0003
[00809] To a solution of ethyl 10'-isopropoxy-9'-(3-methoxypropoxy)-2'-oxo-
2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (45 mg, 0.1 mmol) in MeOH (5 mL) was added LiOH (7.2 mg, 03 mmol) in water (1 mL). After stirring at room temperature for 20 minutes, the reaction was quenched with 1 N HCI to pH 6, then was extracted with 15% IPA in DCM. The organic layer was dried over Na2S04, concentrated and the crude product which was purified by reverse phase reverse phase HPLC (C18, 30- 100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (10.6 mg, 25% yield) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1 H), 7.55 (s, 1 H), 7.35 (s, 1 H), 7.08 (s, 1 H), 4.72 - 4.67 (m, 1 H), 4.1 1 (t, J = 6.3 Hz, 2H), 3.50 (t, J = 6.2 Hz, 2H), 3.28 (s, 2H), 3.26 (s, 3H), 2.68 - 2.59 (m, 2H), 2.05 - 1 .97 (m,4H), 1 .92 - 1 .85 (m, 2H), 1 .28 (s, 3H), 1 .26 (s, 3H). LCMS (ESI) m/z: 428.3 (M + 1)+.
[00810] Example 50
[00811] 9'-lsopentyl-10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolin -3'-carboxylic acid
Figure imgf000171_0001
[00812] Step 1 : 9'-lsopentyl-10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolin -3'-carboxylic acid
Figure imgf000171_0002
[00813] To a solution of ethyl 10'-methoxy-2'-oxo-9'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate (100 mg, 0.21 mmol) in DMF (2 mL) was added 3-methyl-1 -butylboronic acid (1 19 mg, 1 .03 mmol), K2C03 (283 mg, 2.05 mmol), Pd(OAc)2 (10 mg), Ruphos (15 mg) and H20 (0.5 ml). After stirring at 100 °C overnight, the reaction mixture was cooled to room temperature The solvents were removed and the residue was purified by reverse phase HPLC (C18, 30-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (5.4 mg, 7% yield) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1 H), 7.52 (s, 1 H), 7.47 (s, 1 H), 7.23 (s, 1 H), 3.91 (s, 3H), 3.27 (s,2H), 2.66-2.58 (m,4H) , 2.03-1 .97 (m, 2H), 1 .94-1 .84 (m, 2H), 1 .59-1 .53 (m, 1 H), 1 .47-1 .41 (m, 2H), 0.92 (d, J=6.5 Hz, 6H) .LCMS (ESI) m/z: 382.3 (M + 1)+.
[00814] Example 51 [00815] 9'-nsobutylaminoV10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000172_0001
[00816] Step 1 : 9'-(lsobutylaminoV10'-methoxy-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000172_0002
[00817] To a solution of ethyl 10'-methoxy-2'-oxo-9'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate (130 mg, 0.27 mmol) in NMP (2 ml_) was added 2-methylpropan-1 -amine (226 mg, 2.7 mmol). The resulting mixture was heated to 150°C with microwave reactor for 1 hour. After cooling to room temperature, the reaction was partitioned with DCM and H20. The organic layer was separated and washed with brine, dried over Na2S04, filtered and concentrated. The residue was purified by reverse phase reverse phase HPLC (Gilson, C18, 0-100% MeCN in water with 0.1 % formic acid) to afford the title compound (20 mg, 19% yield) as a power. 1H NMR (400 MHz, DMSO-d6) δ 10.43 (t, J = 5.8 Hz, 1 H), 9.80 (s, 1 H), 8.76 (s, 1 H), 7.37 (s, 1 H), 7.01 (s, 1 H), 6.81 (s, 1 H), 3.87 (s, 3H), 3.20 - 3.13 (m, 4H), 2.58 - 2.52 (m, 2H), 2.04 - 1.98 (m,2H), 1.95 - 1.85 (m, 2H), 1.82 - 1.75 (m, 1 H), 0.91 (d, J = 6.7 Hz, 6H). LCMS (ESI) m/z: 383.2(M + 1)+.
[00818] Example 52
[00819] 9'-((Cvclopropylmethyl)amino)-10'-methoxy-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000172_0003
[00820] Step 1 : 9'-((Cvclopropylmethyl)amino)-10'-methoxy-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-al isoquinolinel-3'-carboxylic acid
Figure imgf000173_0001
[00821] To a solution of ethyl 10'-methoxy-2'-oxo-9'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate (130 mg, 0.27 mmol) in NMP (2 mL) was added cyclopropylmethanamine (219 mg, 2.7 mmol). The resulting mixture was heated to 150°C with microwave reactor for 1 hour. After cooling to room temperature, the reaction was partitioned between DCM and H20. The organic layer was separated, washed with brine, dried over Na2S04, filtered and concentrated. The residue was purified by reverse phase HPLC (Gilson, C18, 0-100% MeCN in water with 0.1 % formic acid) to afford the title compound (20 mg, 19% yield) as a power. 1H NMR (400 MHz, DMSO-d6) δ 10.38 (t, J = 5.7 Hz, 1 H), 9.83 (s, 1 H), 8.76 (s, 1 H), 7.37 (s, 1 H), 7.01 (s, 1 H), 6.81 (s, 1 H), 3.87 (s, 3H), 3.22 - 3.19 (m, 2H), 3.16 (s, 2H), 2.56 - 2.51 (m, 2H), 2.04 - 1 .98 (m, 2H), 1 .95 - 1 .85 (m, 2H), 1 .04 - 0.98 (m, 1 H), 0.48 - 0.43 (m, 2H), 0.24 - 0.20 (m, 2H). LCMS (ESI) m/z: 381 .5(M + 1)+.
[00822] Example 53
[00823] 9'-Cvano-10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 - alisoquinolinel-3'-carboxylic acid
Figure imgf000173_0002
[00824] Step 1 : 9'-Cvano-10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000173_0003
[00825] To a solution of ethyl 10'-methoxy-2'-oxo-9'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate (150 mg, 0.31 mmol) in DMF (1 mL) was added Zn(CN)2 (36 mg, 0.31 mmol) and Pd(PPh3)4 (15 mg). After stirring at 1 10 °C overnight, the reaction mixture was cooled down to room temperature. The solvents were removed and the residue was purified by reverse phase reverse phase HPLC (C18, 30-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (28 mg, 27% yield) as a white powder. 1H NMR (400 MHz, DMSO- de) δ 8.85 (s, 1 H), 7.88 (s, 1 H), 7.79 (s, 1 H), 7.74 (s, 1 H), 4.05 (s, 3H), 3.34 (s,2H), 2.70- 2.63 (m,2H) , 2.03-1.97 (m, 2H), 1.92-1.84 (m, 2H) . LCMS (ESI) m/z: 337.1 (M + 1)+.
[00826] Example 54
[00827] 10'-(Dimethylamino)-9'-(furan-3-yl)-2'-oxo-2',7'-dihvdrospiro[cyclobutane-
Figure imgf000174_0001
[00828] Step 1 : Ethyl 9'-(benzyloxy)-10'-(dimethylamino)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000174_0002
[00829] To a mixture of ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate(200 mg, 0.404 mmol), Pd2(dba)3 (20.1 mg, 0.041 mmol),BINAP (37.7 mg, 0.061 mmol), Cs2CO3 (1.06 g,3.232 mmol) in toluene (10 ml), dimethylamine hydrochloride (65.9 mg, 0.808 mmol) was added. The resulting mixture was stirred at 100 °C for 10 h. The mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50ml_x2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel,0~10% MeOH in DCM) to afford the title compound (130 mg, 70.2% yield). LCMS (ESI) m/z: 459.4 (M + 1)+.
[00830] Step 2: Ethyl 10'-(dimethylaminoV9'-hvdroxy-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000174_0003
[00831] A mixture of ethyl 9'-(benzyloxy)-10'-(dimethylamino)-2'-oxo-2',7'- dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (130 mg, 0.283 mmol) and 10% wet Pd/C (100 mg) in MeOH (10 ml_) was stirred under hydrogen atmosphere at room temperature for 1 h.The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was washed with a mixture solvent of PE/EA=1/1 (60 ml_), dried under reduced pressure to afford the title compound (89 mg, 85.4% yield). LCMS (ESI) m/z: 369.3 (M + 1)+.
[00832] Step 3: Ethyl 10'-(dimethylamino)-2'-oxo-9'-(((trifluoromethyl)sulfonyl)oxy)- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000175_0001
[00833] To a stirred mixture of ethyl 10'-(dimethylamino)-9'-hydroxy-2'-oxo-
2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (89 mg, 0.242 mmol) and Cs2C03 (159 mg, 0.484 mmol) in anhydrous DMF (10 ml_), PhNTf2 (103.6 mg, 0.290mmol) was added at 0 °C. The resulting mixture was stirred at room temperature for 1 h. The mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50ml_ x 2). The combined organic layers were washed with brine, dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel, 0-10% MeOH in DCM) to afford the title compound (80 mg, 66% yield). LCMS (ESI) m/z: 501 .3 (M + 1)+.
[00834] Step 4: Ethyl 10'-(dimethylamino)-9'-(furan-3-yl)-2'-oxo-2'.7'-dihvdrospiro [cyclobutane-1 ,6'-pyrid -alisoquinolinel-3'-carboxylate
Figure imgf000175_0002
[00835] To a solution of ethyl 10'-methoxy-2'-oxo-9'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate
[00836] (80 mg, 0.16 mmol) in toluene (5 ml), furan-3-ylboronic acid (35.8 mg, 0.32mmol), Pd(PPh3)4 (18.49 mg, 0.016 mmol), Cs2C03 (157.7 mg,0.48 mmol) and H20 (1 ml) were added. The resulting mixture was stirred at 100 °C for 8 h. The mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50 [00837] ml_x2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography(silica
gel,0~10%MeOH in DCM) to afford the title compound (43 mg, 64.2% yield). LCMS (ESI) m/z: 419.4 (M + 1)+.
[00838] Step 5: 10'-(Dimethylamino)-9'-(furan-3-yl)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000176_0001
[00839] To a solution of ethyl 10'-(dimethylamino)-9'-(furan-3-yl)-2'-oxo-2',7'- dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (43 mg, 0.103 mmol) in MeOH (5 ml), NaOH (32.9 mg, 0.822 mmol) dissolved in H20 (1 ml) was added. The resulting mixture was stirred at room temperature for 2 h. The reaction mixture was acidified to ph~5 with 1 N HCI. Then the mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50ml_x2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by reverse phase reverse phase HPLC (C18 column,30%~100%MeCN in H20, with 0.1 % formic acid in H20) to give 10'-(dimethylamino)-9'-(furan-3-yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid (18.5 mg, 46% yield) as a yellow solid. LCMS (ESI) m/z: 391 .7 (M + 1)+. 1H NMR (400 MHz, DMSO-d6) δ 8.84 (s, 1 H), 8.28 (s, 1 H), 7.78 (t, J = 1 .6 Hz, 1 H), 7.64 (s, 1 H), 7.58 (s, 1 H), 7.49 (s, 1 H), 7.04 (d, J = 1 .1 Hz, 1 H), 3.32 (s, 2H), 2.70 - 2.63 (m, 8H), 2.06 - 2.00 (m, 2H), 1 .94 - 1 .87 (m, 2H).
[00840] Example 55
[00841] 9'-(furan-3-yl)-2'-oxo-10'-((1 .1 .1 -trifluoro-/V-methylmethyl)sulfonamido)-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000176_0002
[00842] Step 1 : Ethyl 9'-(benzyloxyV 10'-(methylaminoV2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000177_0001
[00843] To a mixture of ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (200 mg, 0.404 mmol), ,Pd2(dba)3 (20.1 mg, 0.041 mmol), BINAP (37.7 mg, 0.061 mmol), Cs2CO3 (1 .06 g,3.232 mmol) in toluene (10 ml), methanamine in THF (0.4 ml, 0.808 mmol) was added. The resulting mixture was stirred at 100 °C for 10 h. The mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50ml_x2). The combined organic layer was dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel,0~10% MeOH in DCM) to afford the title compound (160 mg, 89.1 % yield). LCMS (ESI) m/z: 445.4 (M + 1)+.
[00844] Step 2: Ethyl 9'-hvdroxy-10'-(methylamino)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000177_0002
[00845] A mixture of ethyl 9'-(benzyloxy)-10'-(methylamino)-2'-oxo-2',7'- dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (160 mg, 0.360 mmol) and 10% wet Pd/C (100 mg) in MeOH (10 ml_) was stirred under hydrogen atmosphere at room temperature for 1 h.The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was washed with a mixture solvent of PE/EA=1/1 (60 ml_), dried under reduced pressure to afford the title compound (90 mg, 70.5% yield). LCMS (ESI) m/z: 355.9 (M + 1)+.
[00846] Step 3: Ethyl 2'-oxo-10'-((1 .1 .1 -trifluoro-/V-methylmethyl)sulfonamido)-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel- 3'-carboxylate
Figure imgf000177_0003
[00847] To a mixture of ethyl 9'-hydroxy-10'-(methylamino)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (90 mg, 0.254 mmol) and Cs2C03 (333.8 mg, 1.16 mmol) in anhydrous DMF (10 ml_), PhNTf2 (200.4 mg, 0.610 mmol) was added at 0 °C. The resulting mixture was stirred at room temperature for 1 h. The mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50ml_x2). The combined organic layers were washed with brine, dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel, 0-10% MeOH in DCM) to afford the title compound (130 mg, 82.7% yield). LCMS (ESI) m/z: 619.2 (M + 1)+.
[00848] Step 4: Ethyl 9'-(furan-3-yl)-2l-oxo-10'-((1 .1 .1 -trifluoro-N- methylmethyl)sulfonamido)-2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'- carboxylate
Figure imgf000178_0001
[00849] To a solution of ethyl 2'-oxo-10'-((1 ,1 ,1 -trifluoro-/V- methylmethyl)sulfonamido)-9'-(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (130 mg, 0.210 mmol) in toluene (10 ml), furan-3-ylboronic acid (47 mg, 0.420 mmol), Pd(PPh3)4 (24.3 mg, 0.021 mmol), Cs2C03 (413.9 mg,1.26 mmol), H20 (1 ml) were added. The resulting mixture was stirred at 100 °C for 8 h.The mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50ml_x2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel, 0-10% MeOH in DCM) to afford the title compound (76 mg, 67.4% yield). LCMS (ESI) m/z: 537.8 (M + 1)+.
[00850] Step 5: 9'-(Furan-3-yl)-2l-oxo-10'-((1.1.1-trifluoro-N- methylmethyl)sulfonamido)-2',7'-dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'- carboxylic acid
Figure imgf000178_0002
[00851] To a solution of ethyl 9'-(furan-3-yl)-2'-oxo-10'-((1 ,1 ,1-trifluoro-N- methylmethyl)sulfonamido)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'- carboxylate (20 mg, 0.037 mmol) in MeOH (5 ml), NaOH (11 .9 mg, 0.298 mmol) dissolved in H20 (1 ml) was added. The resulting mixture was stirred at room temperature for 2 h. The reaction mixture was acidified to pH = ~5 with 1 N HCI. Then the mixture was diluted with EtOAc (20ml_) and H20 (30 mL), and then extracted with EtOAc (50ml_x2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by reverse phase reverse phase HPLC (C18 column,40%~100%MeCN in H20, with 0.1 % formic acid in H20) to give 9'-(furan-3-yl)-2'-oxo-10'-((1 ,1 ,1 -trifluoro-N- methylmethyl)sulfonamido)-2',7'-dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'- carboxylic acid (5 mg, 26.6 % yield) as a white solid. LCMS (ESI) m/z: 509.3 (M + 1)+. 1H NMR (400 MHz, DMSO-d6) δ 8.86 (s, 1 H), 8.32 (s, 1 H), 8.12 (s, 1 H) 7.88 (t, J = 1 .7 Hz, 1 H), 7.80 (s, 1 H), 7.58 (s, 1 H), 6.91 (d, J = 1.OHz, 1 H), 3.57 (s, 3H), 3.46 (s, 2H) 2.69 - 2.57 (m, 2H), 2.03 - 1.90 (m, 4H).
[00852] Example 56
[00853] 10'-(Azetidin-1-yl)-9'-(furan-3-yl)-2'-oxo-2'.7'-dihvdrospiro[cvclobutane-1.6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000179_0001
[00854] Step 1 : Ethyl 10l-(azetidin-1-yl)-9'-(benzyloxy)-2l-oxo-2l.7l- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000179_0002
[00855] To a mixture of ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate(200 mg, 0.404 mmol),Pd2(dba)3 (20.1 mg, 0.041 mmol), BINAP (37.7 mg,0.061 mmol), Cs2C03 (1.06 g, 3.232 mmol) in toluene (10 ml), azetidine (46.1 mg, 0.808 mmol) was added. The resulting mixture was stirred at 100 °C for 10 h. The mixture was diluted with EtOAc (20ml_) and H20 (30 mL), and then extracted with EtOAc (50mLx2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by column
chromatography (silica gel, 0~10% MeOH in DCM) to afford the title compound (185 mg, 97.3% yield). LCMS (ESI) m/z: 471.4 (M + 1)+.
[00856] Step 2: Ethyl 1 Q'-(azetidin-1 -vD-g'-hvdroxy^'-oxo^'J1- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000180_0001
[00857] A mixture of ethyl 10'-(azetidin-1 -yl)-9'-(benzyloxy)-2'-oxo-2',7'- dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (185 mg, 0.393 mmol) and 10% wet Pt/C (60 mg) in MeOH (10 mL) was stirred under hydrogen atmosphere at room temperature for 1 h.The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was washed with PE/EA=1/1 (60 mL), dried under reduced pressure to afford 135 mg crude ethyl 10'-(azetidin-1 -yl)-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate which was used as is in the next step. LCMS (ESI) m/z: 380.2. (M + 1)+.
[00858] Step 3: Ethyl 1 Q'-(azetidin-1 -yl)-2'-oxo-9'-(((trifluoromethyl)sulfonyl)oxy)-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000180_0002
[00859] To a mixture of ethyl 10'-(azetidin-1 -yl)-9'-hydroxy-2'-oxo-2',7'- dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (135 mg, 0.369 mmol) and TEA (74.5 mg, 0.738 mmol), DMAP (5 mg, 0.037 mmol) in anhydrous DCM (10 mL), PhNTf2 (158.3 mg, 0.443 mmol) was added at 0 °C. The resulting mixture was stirred at room temperature for 1 h. The mixture was diluted with H20 (30 mL), and then extracted with DCM (50 mL x 2). The combined organic layers were washed with brine, dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel, 0-10% MeOH in DCM) to afford the title compound (150 mg, 79.3% yield). LCMS (ESI) m/z: 513.2 (M + 1)+.
[00860] Step 4: Ethyl l O'-fezetidin-l-viyg'-ffuran-S-vn^'-oxo^'J1- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000181_0001
[00861] To a solution of ethyl 10'-(azetidin-1 -yl)-2'-oxo-9'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate(150 mg, 0.293 mmol) in toluene (10 ml), furan-3-ylboronic acid (65.5 mg, 0.585mmol), Pd(PPh3)4 (33.5 mg, 0.029 mmol), Cs2C03 (288.7 mg,0.88 mmol), H20 (1 ml) were added. The resulting mixture was stirred at 100 °C for 8 h. The mixture was diluted with EtOAc (20ml_) and H20 (30 mL), and extracted with EtOAc (50 mL x 2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel,0~10% MeOH in DCM) to afford the title compound (68 mg, 53.9% yield). LCMS (ESI) m/z: 431.4 (M + 1)+.
[00862] Step 5: l O'^Azetidin-l-viyg'-ffuran-S-yn^'-oxo^'J'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000181_0002
[00863] To a solution of ethyl 10'-(azetidin-1-yl)-9'-(furan-3-yl)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (68 mg, 0.158 mmol) in MeOH (5 ml), NaOH (50.5 mg, 1.264 mmol) dissolved in H20 (1 ml) was added. The resulting mixture was stirred at room temperature for 2 h. The reaction mixture was acidified to pH = ~5 with 1 N HCI. The mixture was diluted with EtOAc (20mL) and H20 (30 mL), and then extracted with EtOAc (50 mL x 2). The combined organic solution was dried over anhydrous Na2S04 and concentrated. The residue was purified by reverse phase reverse phase HPLC Gilson (C18 column,30%~100%MeCN in H20,with 0.1 % formic acid in H20) to give 10'-(azetidin-1-yl)-9'-(furan-3-yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid (15 mg, 23.6% yield) as a yellow solid. LCMS (ESI) m/z: 403.8 (M + 1)+. 1H NMR (400 MHz, DMSO-d6) δ 8.84 (s, 1 H), 7.89 (dd, J = 1.4, 0.8 Hz, 1 H), 7.77 (t, J = 1.7 Hz, 1 H), 7.47 (s, 1 H), 7.24 (s, 1 H), 7.08 (s, 1 H), 6.77 (dd, J = 1.8, 0.8 Hz, 1 H), 3.68 (t, J = 7.2 Hz, 4H), 3.26 (s, 2H), 2.69 - 2.61 (m, 2H), 2.17 - 2.10 (m, 2H), 2.06 - 2.00 (m, 2H), 1 .94 - 1 .86 (m, 2H).
[00864] Example 57
[00865] 9'-(Furan-3-ylV10'-(methylamino)-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000182_0001
[00866] Step 1 : Ethyl 9'-(benzyloxy)-10'-(methylamino)-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000182_0002
[00867] To a mixture of ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (200 mg, 0.404 mmol), ,Pd2(dba)3 (20.1 mg, 0.041 mmol), BINAP (37.7 mg,0.061 mmol), Cs2CO3 (1 .06 g,3.232mmol) in toluene (10 ml), methanamine in THF (0.4ml, 0.808 mmol) was added. The resulting mixture was stirred at 100 °C for 10 h. The mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50ml_x2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel, 0-10% MeOH in DCM) to afford the title compound (160 mg, 89.1 % yield). LCMS (ESI) m/z: 445.4 (M + 1)+.
[00868] Step 2: Ethyl 9'-(benzyloxy)-2'-oxo-10'-(2.2.2-trifluoro-/V-methylacetamido)- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000182_0003
[00869] At a 0 °C solution of ethyl 9'-(benzyloxy)-10'-(methylamino)-2'-oxo-
2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (70 mg, 0.157mmol) and TEA (31 .3 mg, 0.31 mmol) in DCM (5 ml), 2,2,2-trifluoroacetic anhydride (39.6 mg, 0.188 mmol) was added. The resulting mixture was stirred at 0 °C for 2 h. The mixture was diluted with H20 (30 mL), and then extracted with DCM (20 mL x 2). The combined organic layers were washed with brine, dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel, 0-5 % MeOH in DCM) to afford the title compound (80 mg, 94.3% yield). LCMS (ESI) m/z: 541 .2 (M + 1)+.
[00870] Step 3: Ethyl 9'-hvdroxy-2'-oxo-10'-(2.2.2-trifluoro-/V-methylacetamidoV2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000183_0001
[00871] A solution of ethyl 9'-(benzyloxy)-2'-oxo-10'-(2,2,2-trifluoro-/V- methylacetamido)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (80 mg, 0.148 mmol) in TFA (10 mL) was stirred at 90 °C for 4 h. The reaction mixture was cooled to room temperature, the solvents were removed under vacuum to give 60mg crude title compound. LCMS (ESI) m/z calcd for C22H2iF3N2O5:450.14. Found: 451 .3 (M + 1)+.
[00872] Step 4: Ethyl 2'-oxo-10'-(2.2.2-trifluoro-/V-methylacetamido)-9'- (((trifluoromethyl) sulfonyl)oxy)-2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel- 3'-carboxylate
Figure imgf000183_0002
[00873] To a solution of ethyl 9'-hydroxy-2'-oxo-10'-(2,2,2-trifluoro-/V- methylacetamido)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (60 mg, 0.133 mmol) in DCM (10 mL) was added N,N-dimethylpyridin-4-amine (1 .6 mg, 0.013 mmol), TEA (26.9 mg, 0.266 mmol) and 1 ,1 ,1 -trifluoro-N-phenyl-N- ((trifluoromethyl)sulfonyl)methanesulfonamide (71 .4 mg, 0.2mmol) under N2 atmosphere. The reaction mixture was stirred at room temperature for 4 h. The solvents were concentrated. The residue was purified by column chromatography using ethyl acatate to afford the title compound (75 mg, 96.8% yield). LCMS (ESI) m/z: 583.2 (M + 1)+.
[00874] Step 5: Ethyl 9'-(furan-3-yl)-2'-oxo-10'-(2.2.2-trifluoro-/V-methylacetamido)- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000184_0001
[00875] To a solution of ethyl 2'-oxo-10'-(2,2,2-trifluoro-/V-methylacetamido)-
9'-(((trifluoromethyl) sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (75 mg, 0.129 mmol) in toluene (5 ml), furan-3-ylboronic acid (28.9 mg, 0.258mmol), Pd(PPh3)4 (15 mg,0.013mmol), Cs2C03 (127mg, 0.387 mmol) and H20 (1 ml) were added. The resulting mixture was stirred at 100 °C for 8 h. Then the mixture was diluted with EtOAc (20ml_) and H20 (30 mL), and then extracted with EtOAc (50ml_x2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel, 0-20 % MeOH in DCM) to afford the title compound (20 mg, 31 % yield). LCMS (ESI) m/z: 501 .3 (M + 1)+.
[00876] Step 6: 9l-(Furan-3-yl)-10l-(methylamino)-2'-oxo-2l.7l- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000184_0002
[00877] To a solution of ethyl 9'-(furan-3-yl)-2'-oxo-10'-(2,2,2-trifluoro-/V- methylacetamido)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (20 mg, 0.04 mmol) in MeOH(5 ml), NaOH (6.4 mg, 0.16 mmol) dissolved in H20 (1 ml) was added. The resulting mixture was stirred at room temperature for 2 h. The reaction mixture was acidified to pH = ~5 with 1 N HCI. Then the mixture was diluted with EtOAc (20ml_) and H20 (30 mL), and then extracted with EtOAc (50mLx2). The combined organic layers were dried over anhydrous Na2S04 and concentrated. The residue was purified by reverse phase reverse phase HPLC Gilson (C18 column, 30%~100%MeCN in H20, with 0.1 % formic acid in H20) to give 9'-(furan-3-yl)-10'-(methylamino)-2'-oxo-2',7'- dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (3.2 mg, 21 .3% yield) as a yellow solid. LCMS (ESI) m/z: 377.4 (M + 1)+. 1H NMR (400 MHz, CDCb) δ 16.06 (s, 1 H), 8.95 (s, 1 H), 7.70 (s, 1 H), 7.58 (t, J = 1 .6 Hz, 1 H), 7.22 (s, 1 H), 7.12 (s, 1 H), 6.91 (s, 1 H), 6.63 (dd, J = 1 .7, 0.7 Hz, 1 H), 3.18 (s, 2H), 2.90 (s, 3H), 2.66 - 2.59 (m, 2H), 2.22 - 2.17 (m, 2H), 2.06 - 1 .95 (m, 2H).
[00878] Example 58 [00879] 9'-Hexyl-10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-
Figure imgf000185_0001
[00881] To a solution of ethyl 10'-methoxy-2'-oxo-9'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate (150 mg, 0.31 mmol) in toluene (2 ml_) was added hexylboronic acid (120 mg, 0.92 mmol) and Pd(PPh3)4 (15 mg), Cs2C03 (802 mg, 2.46 mmol) in H20 (0.5ml_). After stirred at 100 °C overnight, the reaction mixture was cooled to room temperature. The solvents were removed and the residue was purified by reverse phase reverse phase HPLC (C18, 30-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (24 mg, 20% yield) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1 H), 7.52 (s, 1 H), 7.47 (s, 1 H), 7.23 (s, 1 H), 3.91 (s, 3H), 3.26 (s,2H), 2.68-2.57 (m,4H) , 2.03-1.97 (m, 2H), 1.94-1 .84 (m, 2H), 1.58-1 .50 (m, 2H), 1.33-1.24 (m, 6H), 0.86 (t, J=6.8 Hz, 3H) . LCMS (ESI) m/z: 396.3 (M + 1)+.
[00882] Example 59
[00883] 10'-Methoxy-2'-oxo-9'-propyl-2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1- alisoquinolinel-3'-carboxylic acid
Figure imgf000185_0002
[00884] Step 1 : Ethyl 9'-allyl-10'-methoxy-2'-oxo-2'.7'-dihvdrospiro[cvclobutane-1.6'- pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000186_0001
[00885] To a stirred solution of ethyl 10'-methoxy-2'-oxo-9'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate (100 mg, 0.205 mmol) in THF (8 ml) were added CsF (93.45 mg, 0.614 mmol), Pd(Ph3)4 (47.4 mg, 0.04 mmol), and stirred for 30 min at r.t under argon. Allyl boronic acid pinacol ester(103.5 mg, 0.615 mmol) was then added and the reaction mixture was stirred for 12 hr at reflux. The reaction was quenched with deionized H20 and the mixture was extracted with EtOAc. The organic layer was dried over MgS04, concentrated and was purified via silica gel chromatography affording the title compound (48 mg, 62% yield) as a white solid. LCMS (ESI) m/z: 380.4 (M + 1)+.
[00886] Step 2: Ethyl 10'-methoxy-2'-oxo-9'-propyl-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000186_0002
[00887] A solution of ethyl 9'-allyl-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (55.8 mg, 0.147 mmol) and Pd/C (3 mg) in EtOAc was stirred at r.t overnight under H2. The reaction mixture was filtered, and the filtrate was concentrated. The residue was purified to provide ethyl 10'- methoxy-2'-oxo-9'-propyl-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (53.8 mg, 96% yield) as a white solid. LCMS (ESI) m/z: 382.3 (M + 1)+.
[00888] Step 3: 10'-Methoxy-2'-oxo-9'-propyl-2',7'-dihvdrospiro[cvclobutane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000186_0003
[00889] To a solution of ethyl 10'-methoxy-2'-oxo-9'-propyl-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (50 mg, 0.13 mmol) in THF (5 mL) and H20(1 mL) were added LiOH (12.4 mg, 0.52 mmol). After stirring at r.t for 2 hr. the reaction mixture was acidified to pH=5~6 and extracted with EtOAc. The organic layer was dried over Na2S04. The solvent was removed under vacuum and the residue was purified by reverse phase reverse phase HPLC (C18, 0-40% acetonitrile in H20 with 0.1 % formic acid) to provide 10'-methoxy-2'-oxo-9'-propyl-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1-a]isoquinoline]-3'-carboxylic acid (5.04 mg, 1 1.0% yield) as a white solid. LCMS (ESI) m/z: 354.2 (M + 1)+. Ή NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1 H), 7.53 (s, 1 H), 7.48 (s, 1 H) 7.22 (s, 1 H), 3.91 (s, 3H), 3.26 (s, 2H), 2.70 - 2.52 (m, 4H), 2.07 - 1 .96 (m, 2H), 1.94 - 1 .83 (m, 2H), 1.58 (m, 2H), 0.92 (t, J = 7.3 Hz, 3H).
[00890] Example 60
[00891] 9'-(Cvclopropylmethoxy)-2'-oxo-10'-propyl-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000187_0001
[00892] Step 1 : Ethyl 10'-allyl-9'-(cvclopropylmethoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000187_0002
[00893] To a stirred solution of ethyl 10'-bromo-9'-(cyclopropylmethoxy)-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (100 mg, 0.22 mmol) in THF (8 ml) were added CsF (100 mg, 0.66 mmol), Pd(Ph3)4 (46.5 mg, 0.04 mmol), and stirred for 30 min at r.t under argon. Allyl boronic acid pinacol ester (103.5 mg, 0.615 mmol) was then added and the reaction mixture was stirred for 12 hr at reflux. The reaction was quenched with deionized H20 and the reaction mixture was extracted with EtOAc. The organic layer was dried over MgS04, concentrated and the crude material was purified via flash chromatography to afford the title compound (60 mg, 65% yield) as a white solid. LCMS (ESI) m/z calcd for C26H29NO4: 419.21. Found: 420.37 (M + 1)+.
[00894] Step 2: Ethyl 9'-(cvclopropylmethoxy)-2'-oxo-10'-propyl-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000188_0001
[00895] A solution of ethyl 10'-allyl-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (60 mg,0.14 mmol) and Pd/C (3 mg) in EtOAc was stirred at r.t overnight under H2 atmosphere. The reaction mixture was filtered, concentrated and the crude material was purified to provide ethyl 9'- (cyclopropylmethoxy)-2'-oxo-10'-propyl-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (45 mg, 76% yield) as a white solid. LCMS (ESI) m/z: 422.5 (M + 1 )+.
[00896] Step 3: 9'-(Cvclopropylmethoxy)-2'-oxo-10'-propyl-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000188_0002
[00897] To a solution of ethyl 9'-(cyclopropylmethoxy)-2'-oxo-10'-propyl-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (45 mg, 0.107 mmol) in THF (5 mL) and H20 (1 mL) were added LiOH (10.22 mg, 0.427 mmol). After stirred at r.t for 2hr. the reaction mixture was acidified to pH=5~6 and extracted with EtOAc. The organic layer was dried over Na2S04. The solvents were removed under vacuum and the residue was purified by reverse phase reverse phase HPLC (C18, 0-40% acetonitrile in H20 with 0.1 % formic acid) to provide 9'-(cyclopropylmethoxy)-2'-oxo-10'-propyl-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (27 mg, 64.4% yield) as a white solid. LCMS (ESI) m/z: 394.9 (M + 1)+. 1H NMR (400 MHz, DMSO-d6) δ 8.79(s, 1 H), 7.80 (s 1 H), 7.27 (s, 1 H), 7.02 (s, 1 H), 3.96 (d, J=6.8Hz, 2H), 3.31 (s, 2H), 2.65 (s, 1 H), 2.62-2.58 (m, 2H), 2.01 (dd, J = 9.7,5.2 Hz, 2H), 1 .94-1 .85 (m, 2H), 1 .67-1 .57(m, 2H), 1 .35-1 .24 (m, 2H), 0.91 (t, J = 7.4 Hz, 3H), 0.62-0.57 (m, 2H), 0.39-0.34 (m, 2H).
[00898] Example 61
[00899] 9'-Ethyl-10'-methoxy-2'-oxo-2'.7'.1 1 '.1 1 a'-tetrahvdrospiro[cvclobutane-1 .6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000189_0001
[00900] Step 1 : Ethyl 9'-ethyl-10'-methoxy-2'-oxo-2'.7'-dihvdrospiro[cvclobutane-1 .6'- pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000189_0002
[00901] To a solution of ethyl 10'-methoxy-2'-oxo-9'-vinyl-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (80 mg, 0.22 mmol) in MeOH (3 ml_) was added palladium (10 mg, 10% on carbon, ca. 50% water). The resulting mixture was stirred at room temperature under H2 atmosphere (~1 atm) overnight. After removal of palladium by filtration, the filtrate was concentrated and the residue was purified by flash chromatography (silica gel, 0~20% MeOH in DCM) to afford the title compound. (45 mg, 56 % yield) as a light yellow solid. LCMS (ESI) m/z: 368.6 (M + 1)+.
[00902] Step 2: 9'-Ethyl-10'-methoxy-2'-oxo-2'.7'-dihvdrospiro[cvclobutane-1 .6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000189_0003
[00903] To a solution of ethyl 9'-ethyl-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (45 mg, 0.1 1 mmol) in MeOH (3 ml_) was added a solution of NaOH (10 mg, 0.24 mmol) in water (1 ml_). After stirring at room temperature for 20 minutes, the reaction was quenched with 1 N HCI to pH 7, extracted with 15% IPA in DCM. The organic layers were dried over Na2S04, filtered and concentrated. The residue was purified by reverse phase HPLC (Gilson, C18, 30%~100% MeCN in water with 0.1 % formic acid) to afford the title compound (8 mg, 20%) as a white solid. Ή NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1 H), 7.53 (s, 1 H), 7.47 (s, 1 H), 7.25 (s, 1 H), 3.92 (s, 3H), 3.27 (s, 2H), 2.66 - 2.59 (m, 4H), 2.03 - 1 .98 (m, 2H), 1 .94 - 1 .85 (m, 2H), 1 .17 (t, J = 7.5 Hz, 3H). LCMS (ESI) m/z: 340.2 (M + 1)+.
[00904] Example 62 [00905] 9'-(isopropoxymethylV10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000190_0001
[00906] Step 1 : Ethyl 9'-(hvdroxymethyl)-10'-methoxy-2'-oxo-2'.7'- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
[00907] To a solution of ethyl 9'-formyl-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (400 mg, 1 .09 mmol) in THF (10 mL) at 0 °C was added NaBH4 (20 mg, 0.54 mmol) portion-wise. After stirring at 0 °C for 15 minutes, the reaction was quenched with sat'd. NH4CI and extracted with 15% IPA in DCM. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated The residue was used directly in the next step. (200 mg, 50%). LCMS (ESI) m/z: 370.4 (M + 1)+
[00908] Step 2: 9'-(lsopropoxymethyl)-10'-methoxy-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000190_0003
[00909] To a solution of ethyl 9'-(hydroxymethyl)-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate(100 mg, 0.27 mmol) in anhydrous THF (2 mL) at 0 °C was added NaH (32 mg, 0.81 mmol) portion-wise.2- iodopropane (0.91 mL, 0.54 mmol) was added dropwise and the resulting mixture was stirred at room temperature overnight. The reaction mixture was poured into water. LC-MS indicated ester was hydrolyzed, so the above mixture was acidified with 1 N HCI to pH 6, and extracted with 15% IPA in DCM. The organic layer was dried over Na2S04, filtered and concentrated. The residue was purified by reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (2 mg, 1 .9% yield) as a white powder.1H NMR (400 MHz, DMSO-d6) δ 8.83 (s, 1 H), 7.56 (s, 1 H), 7.52 (s, 1 H), 7.41 (s, 1 H), 4.50 (s, 2H), 3.92 (s, 3H), 3.74 - 3.69 (m, 1 H), 3.31 (s, 2H), 2.68 - 2.61 (m, 2H), 2.03 - 1.86 (m, 4H), 1.19 (d, J = 6.1 Hz, 6H).LCMS (ESI) m/z: 384.40 (M + 1)+.
[00910] Example 63
[00911] 9'-(Cvclopropoxymethyl)-10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000191_0001
[00912] Step 1 : Ethyl 10'-methoxy-2'-oxo-9'-((vinyloxy)methyl)-2'.7'- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000191_0002
[00913] The mixture of ethyl 9'-(hydroxymethyl)-10'-methoxy-2'-oxo-2',7'- dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (90 mg, 0.243 mmol), Pd(OAc)2 (5.5 mg, 0.0243 mmol), 1 ,10-phenanthroline(4.4 mg, 0.0243 mmol) and Et3N(24.5 mg, 0.243 mmol) in ethoxyethene (3 mL) and DCM (3 mL) was stirred at 55°C under N2 overnight. The reaction mixture was concentrated and the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to give the title compound (36 mg, 37.5 % yield) as a yellow solid. LCMS (ESI) m/z: 396.2 (M + 1)+.
[00914] Step 2: Ethyl 9'-(cvclopropoxymethyl)-10'-methoxy-2'-oxo-2',7'- dihydrospiro [cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000191_0003
[00915] To a stirred solution of Et2Zn (1 M, 0.36 mL, 0.36 mmol) in DCM(1 mL) at 0°C under N2, was added CH2I2 (96 mg, 0.36 mmol) in DCM (1 mL) dropwise. The mixture was stirred at 0°C for 30 min, then a °C solution of ethyl 10'-methoxy-2'-oxo-9'- ((vinyloxy)methyl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'- carboxylate (36 mg, 0.09 mmol) in DCM (1 mL) was added dropwise and the mixture was stirred at r.t for 2 h. Then the mixture was cooled to 0°C, quenched with H20, and extracted with DCM. The organic layer was washed with brine, dried over Na2S04, concentrated and the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to give the title compound (20 mg, 54% yield) as a yellow solid. LCMS (ESI) m/z: 410.2 (M + 1)+.
[00916] Step 3: 9'-(Cvclopropoxymethvn-10'-methoxy-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000192_0001
[00917] To a solution of ethyl 9'-(cyclopropoxymethyl)-10'-methoxy-2'-oxo-
2',7'-dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate(30 mg, 0.073 mmol) in THF/EtOH/H20(2 mL /1 mL /0.5 mL) was added LiOH (18.5 mg, 0.44 mmol). After stirring at r.t for 1 hr, the resulting mixture was neutralized with 1 N HCI, and extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0- 100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (12.5 mg, 44.8% yield ) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 12.20 (br, 1 H),8.83 (s, 1 H), 7.57 (s, 1 H), 7.52 (s, 1 H),7.37 (s, 1 H), 4.56 (s, 2H),3.92 (s, 3H), 3.43 (t d, J=5.9,3.1 Hz, 1 H), 3.28 (s, 2H) 2.68-2.58(m, 2H), 2.03-1.83(m, 4H), 0.58-0.47 (m, 4H). LCMS (ESI) m/z: 382.3 (M +
[00918] Example 64
[00919] 9'-(Butoxymethyl)-10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolin -3'-carboxylic acid
Figure imgf000192_0002
[00920] Step 1 : Ethyl 9'-(butoxymethyl)-10'-methoxy-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000193_0001
[00921] To a suspension of NaH (72 mg, 1 .78 mmol) in THF (1 mL) at 0°C,was added ethyl 9'-(hydroxymethyl)-10'-methoxy-2'-oxo-2',7'- dihydrospiro [cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylate (220 mg, 0.596 mmol) in THF (2 mL) slowly. The mixture was stirred at r.t for 30 min, then 1 -iodobutane (327 mg, 1 .78 mmol) was added and the mixture was stirred at 60°C for 18 h. The reaction mixture was poured into ice, and extracted with DCM. The organic layer was washed with brine, dried over Na2S04 , concentrated and the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to give the title compound (100 mg, 39.5 % yield) as a yellow solid. LCMS (ESI) m/z: 426.3 (M + 1)+.
[00922] Step 2: 9'-(Butoxymethyl)-10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cvclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000193_0002
[00923] To a solution of ethyl 9'-(butoxymethyl)-10'-methoxy-2'-oxo-2',7'- dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.235 mmol) in THF/EtOH/H20 (2 mL /1 mL /0.5 mL) was added LiOH (59 mg, 1 .41 mmol). After stirring at r.t for 1 hr, the resulting mixture was neutralized with 1 N HCI, and extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0- 100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (22.7 mg, 24.3% yield ) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 12.21 (br, 1 H), 8.83 (s, 1 H), 7.56 (s, 1 H), 7.52 (s, 1 H),7.40 (s, 1 H), 4.50 (s, 2H), 3.92 (s, 3H), 3.52 (t, J=6.3 Hz, 2H), 3.33 (s, 2H) 2.67-2.63 (m, 2H), 2.05-1 .80 (m, 4H), 1 .61 -1 .54 (m, 2H), 1 .43-1 .35(m, 2H), 0.91 (t, J = 7.3 Hz, 3H). LCMS (ESI) m/z: 398.3 (M + 1)+.
[00924] Example 65 [00925] 9'-Butyl-10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -
Figure imgf000194_0001
[00927] To a solution of ethyl 10'-methoxy-2'-oxo-9'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate (150 mg, 0.31 mmol) in DMF (1 mL) was added (n-Bu)4Sn (214 mg, 0.62 mmol),Pd(dppf)CI2 (15 mg), and LiCI (78.3 mg,1 .85 mmol). After stirring at 1 10 °C overnight. The mixture was partitioned between water, ethyl acetate (10 mL) and KF (200 mg) was added to quenched (n-Bu)4Sn. The layers were separated, the aqueous layer was extracted with ethyl acatate (1 OmL x 2), and the combined organic layers were dried over anhydrous sodium sulfate. Solvents were removed and the residue was purified by reverse phase HPLC (C18, 30-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (80 mg, 65 % yield) as a white powder. LCMS (ESI) m/z: 396.3 (M + 1)+.
[00928] Step 2: 9'-Butyl-10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolin -3'-carboxylic acid
Figure imgf000194_0002
[00929] To a solution of ethyl 9'-butyl-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (80 mg, 0.2 mmol) in MeOH (2 mL) was added NaOH (32.4 mg, 0.81 mmol) in H20 (0.5mL). After stirred at room temperature for 2h, the reaction mixture was acidified to pH = 7. Solvents were removed and the residue was purified by reverse phase HPLC (C18, 30-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (19.6 mg, 27 % yield) as a white powder. 1H
NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1 H), 7.52 (s, 1 H), 7.47 (s, 1 H), 7.23 (s, 1 H), 3.91 (s, 3H), 3.27 (s, 2H), 2.67 - 2.57 (m, 4H) , 2.03 - 1.97 (m, 2H), 1.94 - 1.84 (m, 2H), 1.58 - 1.51 (m, 2H), 1.36 - 1.29 (m, 2H), 0.91 (t, J = 7.3 Hz, 3H) . LCMS (ESI) m/z: 368.3 (M + 1)+.
[00930] Example 66
[00931] 9'-(3-EthoxypropylV10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000195_0001
[00932] Step 1 : Ethyl 9'-(3-((tert-butyldimethylsilyl)oxy)prop-1 -vn-1 -ylV10'-methoxy- -2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000195_0002
[00933] To a solution of ethyl 10'-methoxy-2'-oxo-9'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate (250 mg, 0.51 mmol) in DMF (2 mL) was added 3-(tert-butyldimethylsilyloxy)- 1-propyne (437 mg, 2.6 mmol), Pd(dppf)CI2 (15 mg), Cul (91.7 mg, 0.51 mmol), and K2CO3 (496 mg, 3.6 mmol). After stirring at 60 °C for 4h, the reaction mixture was cooled down to room temperature, filtered, and the solvents were removed under vacuum. The residue was purified by column chromatography on silica gel (eluting with DCM:MeOH=15:1) to afford the title compound (255 mg, 98 % yield) as a white powder. LCMS (ESI) m/z: 508.5 (M +
[00934] Step 2: Ethyl 9'-(3-mert-butyldimethylsilvnoxy)propyn-10'-methoxy-2'-oxo- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000195_0003
[00935] To a solution of ethyl 9'-(3-((tert-butyldimethylsilyl)oxy)prop-1 -yn-1- yl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'- carboxylate (255 mg, 0.5 mmol) in MeOH (50 mL) was added Pd/C (26 mg). After stirring at room temperature under a H2 atmosphere (~15 psi) for 48 h, Pd/C was filtered, the solvents were removed under vacuum, to afford the title compound (200 mg, 78 % yield) as a white powder. LCMS (ESI) m/z: 512.4 (M + 1 )+.
[00936] Step 3: Ethyl 9'-(3-hvdroxypropyl)-10'-methoxy-2'-oxo-2'.7'- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000196_0001
[00937] To a solution of ethyl 9'-(3-((tert-butyldimethylsilyl)oxy)propyl)-10'- methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (200 mg, 0.39 mmol) in THF(2 mL) was added TBAF (1 .6 mL, 1 M in THF, 1 .6 mmol). After stirring at room temperature for 2 h, the mixture was partitioned between water and DCM (10 mL). The layers were separated and the aqueous layer was extracted with DCM (10mLx2 ). The combined organic layers were dried over anhydrous sodium sulfate.
Solvents were removed and the residue was purified by reverse phase HPLC (C18, 30- 100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (100 mg, 65 % yield) as a white powder. LCMS (ESI) m/z: 398.3 (M + 1)+.
[00938] Step 4: 9'-(3-Ethoxypropyl)-10'-methoxy-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000196_0002
[00939] To a solution of ethyl 9'-(3-hydroxypropyl)-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.25 mmol) in THF (2 mL) was added NaH (50.3 mg, 1 .3 mmol) and Etl (1 18 mg, 0.76 mmol). After stirring at 0 °C for 4 h, the reaction was quenched with water (2 mL) at 0°C, LC-MS indicated ester was hydrolyzed, then solvents were removed and the residue was purified by reverse phase HPLC (C18, 30-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (6.2 mg, 0.06 % yield) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1 H), 7.53 (s, 1 H), 7.48 (s, 1 H), 7.23 (s, 1 H), 3.91 (s, 3H), 3.42-3.35 (m, 4H), 3.27 (s,2H), 2.67 - 2.60 (m,4H) , 2.02 - 1 .97 (m, 2H), 1 .94 - 1 .87 (m, 2H), 1 .80 - 1 .75 (m, 2H), 1 .1 1 (t, J=7.0 Hz, 3H) . LCMS (ESI) m/z: 398.3 (M + 1)+.
[00940] Example 67 [00941] 10'-Methoxy-2'-oxo-9'-phenethyl-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolin -3'-carboxylic acid
Figure imgf000197_0001
[00942] Step 1 : Ethyl 10'-methoxy-2'-oxo-9'-(phenylethvnyl)-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000197_0002
[00943] To a solution of ethyl 10'-methoxy-2'-oxo-9'-
(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate (120 mg, 0.25 mmol) in DMF (1 ml_) was added phenylacetylene (148 mg, 1 .23 mmol), Pd(dppf)CI2 (60 mg), Cul (47 mg, 0.25 mmol), and K2C03 (238 mg, 1 .7 mmol). After stirring at 60 °C overnight, the reaction mixture was cooled down to room
temperature, filtered, and the solvents were removed under vacuum. The residue was purified by column chromatography on silica gel (eluting with DCM:MeOH=15:1) to afford the title compound (79 mg, 72 % yield) as a white powder. LCMS (ESI) m/z: 440.2 (M + 1)+.
[00944] Step 2: Ethyl 10'-methoxy-2'-oxo-9'-phenethyl-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000197_0003
[00945] To a solution of ethyl 10'-methoxy-2'-oxo-9'-(phenylethynyl)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (79 mg, 0.18 mmol) in MeOH (20 ml_) was added Pd/C (8 mg), and the reaction mixture was stirred under a H2 atmosphere (~15 PSI) at room temperature overnight. The reaction mixture was filtered and the solvents were removed under reduced pressure, to afford the title compound (68 mg, 85 % yield) as a white powder. LCMS (ESI) m/z: 444.4 (M + 1)+.
[00946] Step 3: 10'-Methoxy-2'-oxo-9'-phenethyl-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolin -3'-carboxylic acid
Figure imgf000198_0001
[00947] To a solution of ethyl 10'-methoxy-2'-oxo-9'-phenethyl-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (68 mg, 0.15 mmol) in MeOH (2 mL) was added NaOH (25 mg, 0.61 mmol) in H20 (0.5 mL). After stirring at room temperature for 2 h, the reaction mixture was acidified to pH = 7. Solvents were removed and the residue was purified by reverse phase HPLC (C18, 30-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (28 mg, 45 % yield) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1 H), 7.54 (s, 1 H), 7.50 (s, 1 H), 7.29 - 7.25 (m, 2H), 7.22 - 7.18 (m, 4H), 3.93 (s, 3H), 3.23 (s, 2H), 2.90 - 2.85 (m,4H) , 2.67 - 2.62 (m, 2H), 2.01 - 1 .96 (m, 2H), 1.91 - 1.85 (m, 2H) . LCMS (ESI) m/z: 416.3 (M + 1)+.
[00948] Example 68
[00949] 10'-Bromo-9'-(cvclopropylmethoxy)-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000198_0002
[00950] Step 1 : Ethyl 10'-bromo-9'-(cvclopropylmethoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000198_0003
[00951] To a solution of ethyl 10'-bromo-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (3.0 g, 7.4 mmol) in DMF (50 mL) were added K2C03 (2.08 g, 15.0 mmol) and (bromomethyl)cyclopropane (1 .68 g, 12.5 mmol). After stirring at 80 °C for 24 h, the reaction mixture was cooled to room temperature and quenched with H20 (50 mL), extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine and dried over Na2S04. Solvents were removed under reduced pressure and the residue was purified by silica gel chromatography using DCM/MeOH as an eluent to afford the title compound (2.2 g, 65% yield). LCMS (ESI) m/z: 458.1 / 460.1 (M/M+2)+.
[00952] Step 2: 10'-Bromo-9'-(cvclopropylmethoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000199_0001
[00953] To a solution of ethyl 10'-bromo-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.22 mmol) in EtOH (5 mL) and H20 (1 mL) was added LiOH (15.8 mg, 0.66 mmol) at room temperature The reaction mixture was heated at 65 °C for 30 min. The reaction mixture was acidified to pH ~1 and purified by reverse phase HPLC (C18, 0-30% Acetonitrile in the H20 with 0.1 % formic acid) to afford the title compound (21 .3 mg, yield: 22.6 %) as a white solid. LCMS (ESI) m/z: 430.4/432.4 (M/M+1 )+. 1H NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1 H), 8.29 (s, 1 H), 7.37 (s, 1 H), 7.21 (s, 1 H), 4.04 (d, J = 6.9 Hz, 2H), 3.33 (s, 2H), 2.70 - 2.63 (m, 2H),2.04 - 1 .98 (m, 2H), 1 .91 - 1 .84 (m, 2H), 1 .33 - 1 .28 (m, 1 H), 0.64 - 0.59 (m, 2H), 0.41 - 0.37 (m, 2H).
[00954] Example 69
[00955] 9'-(Cvclopropylmethoxy)-10'-(furan-3-yl)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000199_0002
[00956] Stepl : Ethyl 9'-(cvclopropylmethoxy)-10'-(furan-3-yl)-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000200_0001
[00957] To a solution of ethyl 10'-bromo-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate
[00958] (130 mg, 0.284 mmol) in toluene (5 ml), were added furan-3-ylboronic acid (63.5 mg, 0.568 mmol), Pd(PPh3)4 (32.3 mg, 0.028 mmol), Cs2C03 (279.9 mg,0. 852 mmol), and H20(1drop). The resulting mixture was stirred at 100 °C for 3 h, diluted with EtOAc (20 ml_) and H20 (30 ml_), and then extracted with EtOAc (50 ml_ x 2). The combined organic layer was dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel,0~10% MeOH in DCM) to afford the title compound (80 mg, 63.2% yield) as a yellow solid. LCMS (ESI) m/z: 446.3 (M + 1)+.
[00959] Step 2: 9'-(CvclopropylmethoxyV10'-(furan-3-vn-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000200_0002
[00960] Sodium hydroxide (28.7 mg, 0.718mmol) dissolved in H20 (1 ml) was added to a solution of ethyl 9'-(cyclopropylmethoxy)-10'-(furan-3-yl)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (80 mg, 0.179 mmol) in MeOH (5 ml).. The resulting mixture was stirred at room temperature for 2 h and then acidified to ph~5 with 1 N HCI. The mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and the aqueous layer was extracted with EtOAc (50ml_x2). The combined organic layer was dried over anhydrous Na2S04 and concentrated. The residue was purified by Gilson (C18 column,50-100%MeCN in H20, with 0.1 % formic acid in H20) to give 9'- (cyclopropylmethoxy)-10'-(furan-3-yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1- a]isoquinoline]-3'-carboxylic acid
[00961] (26.9 mg, 36% yield) as a white solid. LCMS (ESI) m/z: 418.3 (M + 1)+. Ή NMR (400 MHz, DMSO-d6) δ 8.81 (s, 1 H), 8.29 (d, J = 0.7 Hz, 1 H), 8.20 (s, 1 H), 7.76 (t, J = 1 .7 Hz, 1 H), 7.58 (s, 1 H), 7.42 (d, J = 1 .3 Hz, 1 H), 7.18 (s, 1 H), 4.07 (d, J = 7.2 Hz, 2H), 3.36 (s, 2H), 2.69 - 2.62 (m, 2H), 2.06 - 2.00 (m, 2H), 1 .93 - 1 .86 (m, 2H), 1 .44 - 1 .38 (m, 1 H), 0.69 - 0.64 (m, 2H), 0.44 - 0.39 (m, 2H).
[00962] Example 70
[00963] 10'-Cvclopropyl-9'-(cvclopropylmethoxy)-2'-oxo-2',7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000201_0001
[00964] Step 1 : Ethyl 10'-cvclopropyl-9'-(cvclopropylmethoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000201_0002
[00965] To a solution of ethyl 10'-bromo-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (200 mg, 0.44 mmol), were added cyclopropylboronic acid (75.7 mg, 0.88 mmol), Pd(PPh3)4 (50 mg, 0.044 mmol), Cs2C03 (429 mg, 0.33 mmol), toluene (3 ml_) and water (1 ml_), and the reaction mixture was stirred overnight at 85 °C. The mixture was cooled to r.t and extracted with EtOAc. The combined organic layer was washed with brine, and concentrated to afford the title compound (184 mg, 100% yield). LCMS (ESI) m/z: 420.1/422.1 (M/M+2)+.
[00966] Step 2: 10'-Cvclopropyl-9'-(cvclopropylmethoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000201_0003
[00967] Lithium hydroxide (32 mg, 1 .32 mmol) was added to a solution of ethyl 10'-cyclopropyl-9'-(cyclopropylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylate (184 mg, 0.44 mmol) in EtOH (5 ml_) and H20 (1 ml_). The reaction mixture was heated to 65 °C for 2 h and then acidified to pH ~1 . Water
(40 ml_) was added and the mixture was extracted with EtOAc (2x). The combined organic layer was washed with brine, and then concentrated. The residue was purified by preparative TLC, and further purified by reverse phase HPLC (C18, 0-90% Acetonitrile in H20 with 0.1 % formic acid) to afford the title compound (7.8 mg, 4.5% yield). LCMS (ESI) m/z: 392.3 (M + 1)+. Ή NMR (400 MHz, DMSO-d6) δ 8.77 (s, 1 H), 7.36 (s, 2H), 7.01 (s, 1 H), 3.98 (d, J = 6.8 Hz, 2H), 3.29 (s, 2H), 2.65 - 2.59 (m, 2H), 2.18 - 2.14 (m, 1 H),2.02 - 1 .97 (m, 2H), 1 .92 - 1 .85 (m, 2H), 1 .33 - 1 .29 (m, 1 H), 0.93 - 0.86 (m, 4H), 0.63 - 0.58 (m, 2H), 0.40 - 0.37 (m, 2H).
[00968] Example 71
[00969] 1 ',8'-Dibromo-10'-cvclopropyl-9'-(cvclopropylmethoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000202_0001
[00970] Step 1 : 1 ',8'-Dibromo-10'-cvclopropyl-9'-(cvclopropylmethoxy)-2'-oxo-2',7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000202_0002
[00971] To a solution of 10'-cyclopropyl-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (10 mg,
0.026mmol) in AcOH (2 mL) was added NBS (9.2 mg, 0.052 mmol). The reaction mixture was stirred at 65 °C for 3 h. After TLC showed little title compound in the reaction mixture, AcOH was concentrated and then dissolved in EtOH, The solution was purified by reverse phase HPLC (C18, 0-30% MeCN in H20 with 0.1 % formic acid) to afford the title compound (3.9 mg,36% yield) as a white solid. LCMS (ESI) m/z: 548.1/550.1/552.1 (M/M+2/M+4)+ . 1H NMR (400 MHz, CDCI3) δ 15.21 (s, 1 H), 8.94 (s, 1 H), 7.78 (s, 1 H), 3.97 (d, J = 7.2 Hz, 2H), 3.35 (s, 2H), 2.56 (s, 2H), 2.36 - 2.31 (m, 1 H), 2.05 - 1 .96 (m, 4H), 1 .44 - 1 .39 (m, 1 H), 1 .12 - 1 .08 (m, 2H), 0.79 - 0.76 (m, 2H), 0.70 - 0.66 (m, 2H), 0.42 - 0.38 (m, 2H).
[00972] Example 72
[00973] 9'-(Cvclopropylmethoxy)-10'-isopropyl-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000203_0001
[00974] Step 1 : Ethyl 9'-(cvclopropylmethoxyV2'-oxo-10'-fprop-1 -Θη-2-νΠ-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000203_0002
[00975] To a solution of ethyl 10'-bromo-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (200 mg, 0.44 mmol) and 4,4,5,5-tetramethyl-2-(prop-1 -en-2-yl)-1 ,3,2-dioxaborolane (148 mg, 0.88 mmol) in dioxane (3 mL) and water (2 mL) were added Pd(PPh3)4 (50 mg, 0.044 mmol) and Cs2C03 (429 mg, 1 .32 mmol). The reaction mixture was stirred at 85 °C under N2 for 2 h. The reaction mixture was allowed to reach room temperature, water (1 mL) was added, and the solution was extracted twice with EtOAc. The combined EtOAc layers were washed with brine and concentrated to afford the crude title compound (180 mg, crude) which was used in the next step without future purification. LCMS (ESI) m/z: 420.3 (M + 1)+.
[00976] Step 2: Ethyl 9'-(cvclopropylmethoxy)-10'-isopropyl-2'-oxo-2',7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000203_0003
[00977] To a solution of ethyl 9'-(cyclopropylmethoxy)-2'-oxo-10'-(prop-1 -en-
2-yl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (180 mg, 0.43 mmol) in EtOH (50 mL) was added Pd-C (36 mg, 20% w/w). The mixture was stirred at room temperature under a H2 atmosphere (15 psi) for 48 h. The reaction mixture was filtered and the filtrate was concentrated to afford the crude title compound (60 mg, crude). LCMS (ESI) m/z: 422.5 (M + 1)+.
[00978] Step 3: 9'-(Cvclopropylmethoxy)-10'-isopropyl-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000204_0001
[00979] To a solution of ethyl 9'-(cyclopropylmethoxy)-10'-isopropyl-2'-oxo-
2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (60 mg, 0.14 mmol) in EtOH (5 ml_) and H20 (1 mL) was added LiOH (13.4 mg, 0.56 mmol) at room temperature The reaction mixture was heated to 65 °C for 30 min. The reaction mixture was acidified to pH = ~4 with 1 N HCI and was purified by reverse phase HPLC (C18, 0-30% Acetonitrile in H20 with 0.1 % formic acid) to afford the title compound (18 mg, 33% yield) as a white solid. LCMS (ESI) m/z: 394.2 (M + 1)+. 1H NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1 H), 7.76 (s, 1 H), 7.34 (s, 1 H), 7.02 (s, 1 H), 3.97 (d, J = 6.8 Hz, 2H), 3.30 (s, 2H), 3.28 - 3.26 (m, 1 H), 2.67 - 2.61 (m, 2H), 2.04 - 1.99 (m, 2H), 1.93 - 1.86 (m, 2H), 1 .32 - 1.29 (m, 1 H), 1.25 (d, J = 7.0 Hz, 6H), 0.62 - 0.58 (m, 2H), 0.39 - 0.35 (m, 2H).
[00980] Example 73
[00981] 9'-(Cvclopropylmethoxy)-1 Q'-(1 -methyl-1 H-pyrazol-3-yl)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000204_0002
[00982] Step 1 : Ethyl 9'-(cvclopropylmethoxy)-10'-(1 -methyl-1 H-pyrazol-3-yl)-2'-oxo- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000204_0003
[00983] To a stirred solution of ethyl 10'-bromo-9'-(cyclopropylmethoxy)-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (100 mg, 0.21 mmol) in DME (2 ml) and water (0.5 ml) were added Na2C03 (44 mg, 0.42 mmol) and Pd(dppf)CI2 (8 mg, 0.01 1 mmol), and the reaction mixture was stirred for 30 min at r.t under Ar. (1-methyl-1 H-pyrazol-3-yl)boronic acid (52 mg, 0.42 mmol) was then added and the reaction mixture was stirred for 12 hr at reflux. The reaction was quenched at room temperature by addition of deionized H20. The solution was extracted with EtOAc and the combined organic layer was dried over MgS04, filtered, and concentrated in vacuo. Silica gel chromatography afforded the title compound (50 mg, 49% yield) as a white solid. LCMS (ESI) m/z: 460.4 (M + 1)+.
[00984] Step 2: 9'-(CvclopropylmethoxyV 10'-d -methyl-1 H-pyrazol-3-yr)-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000205_0001
[00985] Lithium hydroxide (12 mg, 0.50 mmol) was added to a solution of ethyl 9'-(cyclopropylmethoxy)-10'-(1 -methyl-1 H-pyrazol-3-yl)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (50 mg, 0.10 mmol) in THF (2 mL) and H20 (1 mL). After stirring at r.t for 1 h, the reaction mixture was acidified to pH=4~5 and extracted with EtOAc. The organic layer was dried over Na2S04. Solvents were removed under vacuum and the residue was purified by reverse phase HPLC (C18, 0- 40% acetonitrile in H20 with 0.1 % formic acid) to provide 9'-(cyclopropylmethoxy)-10'-(1 - methyl-1 H-pyrazol-3-yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylic acid (28 mg, 59% yield) as a white solid. LCMS (ESI) m/z: 432.3 (M + 1)+. 1H NMR (400 MHz, DMSO-d6) 68.82 (s, 1 H), 8.34 (s, 1 H), 7.76 (d, J = 2.1 Hz, 1 H), 7.19 (s, 1 H), 7.08 (s, 1 H), 6.86 (d, J = 2.1 Hz, 1 H), 4.05 (d, J = 7.0 Hz, 2H),3.93 (s, 3H), 3.35 (s, 2H), 2.70 - 2.03 (m, 2H), 2.07-2.63 (m, 2H), 1 .94 - 1 .87 (m, 2H), 1 .38 - 1 .34 (m, 1 H), 0.65- 0.62 (m, 2H), 0.41 - 0.39 (m, 2H).
[00986] Example 74
[00987] 9'-(Cvclopropylmethoxy)-10'-(3-hvdroxypropyl)-2'-oxo-2',7'- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000205_0002
[00988] Step 1 : Ethyl 10'-(3-((tert-butyldimethylsilyl)oxy)prop-1 -yn-1 -yl)-9'- (cyclopropylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate
Figure imgf000206_0001
[00989] To a solution of ethyl 10'-bromo-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (250 mg, 0.54 mmol) in DMF (1 ml_) was added 3-(tert-butyldimethylsilyloxy)-1 -propyne (463 mg, 2.7 mmol), Pd(pph3)CI 2 (125 mg), K2CO3 (525 mg, 3.8 mmol), and Cul (103.4 mg, 0.54 mmol). After stirring at 60 °C for 4 h, the reaction mixture was cooled down to room temperature and the solvents were removed under reduced pressure. The residue was purified by column chromatography on silica gel (eluting with DCM:MeOH=10:1 ) to afford the title compound (287 mg, 97 % yield) as a white powder. LCMS (ESI) m/z: 548.3 (M + 1)+.
[00990] Step 2: Ethyl 10'-(3-((tert-butyldimethylsilyl)oxy)propyl)-9'- (cvclopropylmethoxy)-2'-oxo-2',7'-dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'- carboxylate
Figure imgf000206_0002
[00991] To a solution of ethyl 10'-(3-((tert-butyldimethylsilyl)oxy)prop-1 -yn-1 - yl)-9'-(cyclopropylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (287 mg, 0.52 mmol) in MeOH (50 mL) was added Pd/C (290 mg), and the reaction mixture was purged with H2(3x). After stirring at room temperature under a H2 atmosphere (15 psi) for 48 h, the reaction mixture was filtered, and the solvents were removed under reduced pressure, to afford the title compound (220 mg, 77 % yield) as a white powder. LCMS (ESI) m/z: 552.5 (M + 1 )+.
[00992] Step 3: Ethyl 9'-(cvclopropylmethoxy)-10'-(3-hvdroxypropyl)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000207_0001
[00993] To a solution of ethyl 10'-(3-((tert-butyldimethylsilyl)oxy)propyl)-9'-
(cyclopropylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (220 mg, 0.40 mmol) in THF (4 mL) was added TBAF (1.6 mL, 1 M in THF, 1 .6 mmol). After stirring at room temperature for 4 h, the solvents were removed under reduced pressure, and the residue was purified by prep-TLC to afford the title compound (120 mg, 69 % yield) as a white powder. LCMS (ESI) m/z: 438.4 (M + 1)+.
[00994] Step 4: 9'-(Cvclopropylmethoxy)-10'-(3-hvdroxypropyl)-2'-oxo-2',7'- dihvdrospiro[cvclobutan -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000207_0002
[00995] To a solution of ethyl 9'-(cyclopropylmethoxy)-10'-(3-hydroxypropyl)-
2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (120 mg, 0.27 mmol) in MeOH (2 mL) was added NaOH (26 mg, 1.1 mmol) in H20 (0.5 mL). After stirring at room temperature for 2 h, the reaction mixture was acidified to pH = 7. Solvents were removed and the residue was purified by reverse phase HPLC (C18, 30-100% MeCN in H2O with 0.1 % formic acid) to afford the title compound (73.8 mg, 67 % yield) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1 H), 7.79 (s, 1 H), 7.27 (s, 1 H), 7.02 (s, 1 H), 4.47 (t, J = 5.2 Hz, 1 H), 3.96 (d, J = 6.8 Hz, 2H), 3.44-3.40 (m, 2H), 3.30 (s, 2H), 2.68 - 2.62 (m, 4H), 2.04 - 1.99 (m, 2H), 1.92 - 1.86 (m,2H) , 1.78 - 1.72 (m, 2H), 1 .30 - 1 .26 (m, 1 H), 0.62 - 0.57 (m, 2H), 0.39 - 0.35 (m, 2H) . LCMS (ESI) m/z: 410.3(M + 1)+.
[00996] Example 75
[00997] 10'-(cyclopent-1 -en-1 -yl)-9'-(cvclopropylmethoxy)-2'-oxo-2',7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000207_0003
[00998] Stepl : Ethyl 1 Q'-(cvclopent-1 -en-1 -yl)-9'-(cvclopropylmethoxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000208_0001
[00999] To a solution of ethyl 10'-bromo-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (280 mg, 0.610 mmol) in toluene (10 ml), were added 2-(cyclopent-1 -en-1 -yl)-4,4,5,5-tetramethyl-1 ,3,2- dioxaborolane (237.6 mg, 1 .22 mmol), Pd(PPh3)4 (34.7 mg, 0.06mmol), Cs2CO3 (601.2 mg,1.83 mmol), and H20 (1 drop). The resulting mixture was stirred at 100 °C for 3 h. The mixture was diluted with EtOAc (20ml_) and H2O (30 ml_), and then extracted with EtOAc (50 mL x 2). The combined organic solution was dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel, 0-10% MeOH in DCM) to afford ethyl 10'-(cyclopent-1 -en-1 -yl)-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (200 mg, 73.6% yield). LCMS (ESI) m/z: 446.3 (M + 1)+.
[001000] Step 2: 1 Q'-(Cvclopent-1 -en-1 -yl)-9'-(cvclopropylmethoxy)-2'-oxo- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000208_0002
[001001] To a solution of ethyl 10'-(cyclopent-1 -en-1 -yl)-9'- (cyclopropylmethoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (200 mg, 0.449 mmol) in MeOH(10 ml), was added NaOH (71 .8 mg,
1.795mmol) dissolved in H20(1 ml). The resulting mixture was stirred at room temperature for 2 h. The reaction mixture was acidified to ph~5 with 1 N HCI. Then the mixture was diluted with EtOAc (20ml_) and H20 (30 mL), and extracted with EtOAc (50ml_x2). The combined organic solution was dried over anhydrous Na2S04 and concentrated. The residue was purified by Gilson (C18 column, 50%-100% MeCN in H20, with 0.1 % formic acid in H20) to give 10'-(cyclopent-1-en-1-yl)-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylic acid [001002] (118.7 mg, 63.3% yield) as a white solid. LCMS (ESI) m/z: 418.3 (M
+ 1)+. 1H NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1 H), 7.75 (s, 1 H), 7.33 (s, 1 H), 7.10 (s, 1 H), 6.63 (s, 1 H), 4.00 (d, J = 7.1 Hz, 2H), 3.33 (s, 2H), 2.83 (t, J = 6.5 Hz, 2H), 2.68 - 2.62 (m, 2H), 2.56 - 2.52 (m, 2H), 2.03 - 1 .98 (m, 2H), 1 .93 - 1.86 (m, 4H), 1.34 - 1 .30 (m, 1 H), 0.65 - 0.60 (m, 2H), 0.39 - 0.36 (m, 2H).
[001003] Example 76
[001004] 10'-Cvclopentyl-9'-(cvclopropylmethoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000209_0001
[001005] Step 1 : 10'-Cvclopentyl-9'-(cvclopropylmethoxyV2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000209_0002
[001006] A mixture of 10'-(cyclopent-1 -en-1 -yl)-9'-(cyclopropylmethoxy)-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylic acid (100 mg, 0.238 mmol) and10% wet Pd/C (100 mg) in MeOH (10 mL) was stirred under a hydrogen atmosphere (1 atm) at r. t. for 1 h.The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by Gilson (C18 column, 50%-100%MeCN in H20, with 0.1 % formic acid in H20) to give 10'-cyclopentyl-9'-(cyclopropylmethoxy)-2'-oxo- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (29.6 mg, 29.7% yield) as a white solid. LCMS (ESI) m/z: 420.3 (M + 1)+. Ή NMR (400 MHz, DMSO- d6) δ 8.79 (s, 1 H), 7.77 (s, 1 H), 7.32 (s, 1 H), 7.02 (s, 1 H), 3.96 (d, J = 6.9 Hz, 2H), 3.30 (s, 2H), 3.28 - 3.22 (m, 1 H), 2.68 - 2.60 (m, 2H), 2.03 - 1.94 (m, 4H), 1.92 - 1 .86 (m, 2H), 1.83 - 1.77 (m, 2H), 1.73 - 1.62 (m, 4H), 1.30 - 1.27 (m, 1 H), 0.62 - 0.58 (m, 2H), 0.38 - 0.34 (m, 2H).
[001007] Example 77
[001008] 10'-Methyl-9'-(1 -methyl-1 H-pyrazol^-vD^'-oxo^'J'- dihydrospirofcyclohexane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000210_0001
[001009] Step 1 : Ethyl 9'-(benzyloxy)-10'-methyl^'-oxo^'J1- dihvdrospiro[cvclohexane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000210_0002
[001010] To a solution of ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (500 mg, 0.96 mmol) in toluene (4 mL) was added methyboronic acid (172 mg, 0.2.87 mmol), Pd(PPh3)4 (50 mg), Cs2C03 (1.6 g, 4.8 mmol) and H20 (1 mL). After stirring at 85 °C for 6 h, the reaction mixture was cooled down to room temperature, and the solvents were removed under vacuum to give the crude product which was purified by flash chromatography (silica gel, 0-50% DCM in MeOH) to afford the title compound (440 mg, 100 % yield). LCMS (ESI) m/z: 458.4 (M + 1)·
[001011] Step 2: Ethyl 9'-hvdroxy-10'-methyl^'-oxo^'J1- dihydrospirofcyclohexane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000210_0003
[001012] A solution of ethyl 9'-(benzyloxy)-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (440 mg, 0.96 mmol) in TFA (4 mL) was stirred at 60 °C for 3 h. The reaction mixture was cooled to room temperature and the solvents were removed under reduced pressure to give the desired product (320 mg, 91 % yield). LCMS (ESI) m/z: 368.3 (M + 1).
[001013] Step 3: Ethyl 10'-methyl-2'-oxo-9'-(((trifluoromethyl)sulfonyl)oxy)- 2',7'-dihvdrospiro[cyclohexane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000211_0001
[001014] To a solution of ethyl 9'-hydroxy-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (160 mg, 0.435 mmol) in DCM (10 mL) was added N,N-dimethylpyridin-4-amine (5 mg, 0.044 mmol), TEA (133 mg, 1.32 mmol) and 1 ,1 ,1-trifluoro-N-phenyl-N-
((trifluoromethyl)sulfonyl)methanesulfonamide (233 mg, 0.65 mmol) under N2 atmosphere. The reaction mixture was stirred at room temperature for 4 h. The solvents were concentrated. The residue was purified by column chromatography using EtOAc to afford the title compound (200 mg, 92% yield). LCMS (ESI) m/z: 500.3 (M + 1)+.
[001015] Step 4: Ethyl 10'-methyl-9'-(1 -methyl-1 H-pyrazol^-vD^'-oxo^'J1- dihydrospirofcyclohexa -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000211_0002
[001016] To a solution of ethyl 10'-methyl-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate (90 mg, 0.180 mmol) in toluene (5 ml), 1-methyl-4-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)-1 H-pyrazole (74.9 mg, 0.360mmol), Pd(PPh3)4 (20.8 mg,0.018mmol), Cs2C03 (177.4 mg,0.540mmol), and H20 (1drop) were added. The resulting mixture was stirred at 100 °C for 8 h. Then the mixture was diluted with EtOAc (20ml_) and H20 (30 mL), and then extracted with EtOAc (50 mLx2). The combined organic solution was dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel, 0-10% MeOH in DCM) to afford the title product (41 mg, 52.8% yield) as a white solid. LCMS (ESI) m/z: 432.3 (M + 1)+.
[001017] Step 5: 10'-Methyl-9'-(1 -methyl-1 H-pyrazol^-vD^'-oxo^'J'- dihydrospirofcyclohexane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000212_0001
[001018] To a solution of ethyl 10'-methyl-9'-(1 -methyl- 1 H-pyrazol-4-yl)-2'-oxo- 2',7'-dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (41 mg, 0.095 mmol) in MeOH (5 ml) was added NaOH (30.4 mg, 0.761 mmol) dissolved in H20( 1 ml). The resulting mixture was stirred at room temperature for 2 h. The reaction mixture was acidified to ph~5 with 1 N HCI. Then the mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50ml_x2). The combined organic solution was dried over anhydrous Na2S04 and concentrated. The residue was purified by Gilson (C18
column,30%-100% MeCN in H20, with 0.1 % formic acid in H20) to give 10'-methyl-9'-(1- methyl-1 /-/-pyrazol-4-yl)-2'-oxo-2',7'-dihydrospiro [cyclohexane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid (13 mg, 33.9% yield) as a light yellow solid. LCMS (ESI) m/z: 404.9 (M + 1)+. Ή NMR (400 MHz, DMSO-d6) δ 8.85 (s, 1 H), 8.10 (s, 1 H), 7.99 (s, 1 H), 7.82 (s, 1 H), 7.56 (s, 1 H), 7.44 (s, 1 H), 3.91 (s, 3H), 3.29 (s, 2H), 2.47 (s, 3H), 2.01 - 1 .93 (m, 3H), 1.82 - 1 .77 (m, 2H), 1.69 - 1 .63 (m, 3H), 1.60 - 1.52 (m, 2H).
[001019] Example 78
[001020] 9'-lsobutoxy-10'-methyl-2'-oxo-2',7'-dihvdrospiro[cyclohexane-1 ,6'- pyrido[2,1 -alisoquinolin -3'-carboxylic acid
Figure imgf000212_0002
[001021] Step 1 : Ethyl 9'-isobutoxy-10'-methyl^'-oxo^'J1- dihydrospirofcyclohexa -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000212_0003
[001022] To a solution of ethyl 9'-hydroxy-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (110 mg, 0.3 mmol) in MeCN (1 ml_) was added 1-bromo-2-methyl propane (61.5 mg, 0.45 mmol) and K2C03 (83 mg, 0.6 mmol) . After stirring at 65 °C for 5 h, the reaction mixture was cooled down to room temperature, the solvents were removed under vacuum, and the residue was purified by prep-TLC to provide the desired product (83 mg, 65 % yield). LCMS (ESI) m/z: 424.4 (M + 1)+.
[001023] Step 2: g'-lsobutoxy-I O'-methyl^'-oxo^'J1- dihydrospirofcyclohexa -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000213_0001
[001024] To a solution of ethyl 9'-isobutoxy-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclohexane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (83 mg, 0.2 mmol) in THF (2 mL) was added LiOH (19 mg, 0.8 mmol) and H20 (0.5 ml). After stirring at room temperature for 2 h, the reaction mixture was acidified to pH = 7. Solvents were removed and the residue was purified by reverse phase HPLC (C18, 30-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (29.2 mg, 37 % yield) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 8.81 (s, 1 H), 7.88 (s, 1 H), 7.29 (s, 1 H), 7.08 (s, 1 H), 3.85 (d, J = 5.7 Hz, 2H), 3.28 (s, 2H), 2.21 (s, 3H), 2.10-2.06 (m, 1 H), 1 .98 - 1 .89 (m, 2H), 1.81 - 1.74 (m, 2H), 1.70 - 1.54 (m, 5H), 1.33 - 1.28 (m, 1 H), 1.02(d, J = 6.3 Hz, 6H). LCMS (ESI) m/z: 396.3 (M + 1)+.
[001025] Example 79
[001026] 9'-(Cvclobutylmethoxy)-10'-methyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000213_0002
[001028] Under a stream of nitrogen, ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (5.0 g, 10 mmol), methylboronic acid (2.4 g, 40 mmol), Pd(Ph3)4 (5.8 g, 5 mmol), Cs2C03 (13 g, 40 mmol), toluene (40 mL) and water (2 mL) were placed into a three-necked flask of 200 mL. The reaction mixture was stirred overnight at 100 °C and then added to water (100 mL), and extracted with ethyl acetate. The combined organic layer was dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to provide an oily liquid. This residue was purified by silica gel chromatography (hexane/ethyl acetate as eluent) to afford the title compound (3.4 g, 79% yield), LCMS (ESI) m/z: 430.2 (M + 1)+.
[001029] Step 2: Ethyl 9'-hvdroxy-10'-methyl^'-oxo^'J1- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000214_0001
[001030] A solution of ethyl 9'-(benzyloxy)-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (3.4 g, 8 mmol) in TFA (40 mL) was stirred at 65 °C. The mixture was concentrated and the residue was dissolved in DCM and washed with brine, dried over MgSC and concentrated. The residue was purified by silica gel chromatography to afford the title compound (2.15 g, 75% yield). LCMS (ESI) m/z: 340.1 (M + 1)+.
[001031] Step 3: Ethyl 9'-(cvclobutylmethoxy)-10'-methyl-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000214_0002
[001032] To a solution of ethyl 9'-hydroxy-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (90 mg, 0.26 mmol) in DMF (2 mL) was added K2C03 (73.1 mg, 0.53 mmol) and (bromomethyl)cyclobutane (79.0 mg, 0.53mmol). The reaction mixture was stirred at 90°C for 2 h. Then the reaction was extracted with DCM twice and the combined organic phase was washed with brine, dried over Na2S04, filtered and concentrated. The residue was purified by flash chromatography (silica gel, 0-40% CH3OH in DCM) to afford the title compound (60 mg,
55.6% yield). LCMS (ESI) m/z: 408.4 (M + 1)+. [001033] Step 4: 9'-(Cvclobutylmethoxy)-10'-methyl-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000215_0001
[001034] To a solution of ethyl 9'-(cyclobutylmethoxy)-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (60.5 mg, 0.15 mmol) in MeOH (3ml_) was added NaOH (17.8 mg, 0.45 mmol) in water (1 mL). The resulting mixture was stirred at room temperature for 30 mins and then acidified with 1 N HCI to pH = 6. The reaction mixture was extracted with 15% IPA in DCM twice and the combined organic layer was dried over Na2S04, filtered and concentrated. The residue was purified by reverse phase HPLC (Gilson, Ci8, 30%-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound as a white solid (7 mg, 12% yield). LCMS (ESI) m/z: 380.8 (M + 1)+. 1H NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1 H), 7.85 (s, 1 H), 7.25 (s, 1 H), 7.04 (s, 1 H), 4.07 (d, J = 6.4 Hz, 2H), 3.33 (s, 2H), 2.80 - 2.75 (m, 1 H), 2.67 - 2.60(m, 2H), 2.20 (s, 3H), 2.13 - 2.08 (m, 2H), 2.03 - 1.99 (m, 2H), 1.94 - 1.87 (m, 6H).
[001035] Example 80
[001036] 9'-(2-Cvclopropylethoxy)-10'-methyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000215_0002
[001037] Step 1 :Ethyl 9'-(2-cvclopropylethoxyV10'-methyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000215_0003
[001038] A Schlenk tube was charged with ethyl 9'-hydroxy-10'-methyl-2'-oxo- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.29 mmol), cyclopropylmethyl 4-methylbenzenesulfonate (84 mg, 0.35 mmol) and CH3CN (2 mL). K2C03 (48 mg, 0.35 mmol) was added and the reaction mixture was stirred for 6 h at 85°C. After cooling to room temperature, the inorganic salt was filtered off and ethyl acetate was added. The organic layer and washed with brine. After drying with anhydrous magnesium sulfate, the solvent was evaporated, and the residue was purified by column chromatography to afford the title compound (65 mg, 55% yield). LCMS (ESI) m/z: 408.4 (M + 1 )+.
[001039] Step 2: 9'-(2-CvclopropylethoxyV10'-methyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000216_0001
[001040] To a solution of ethyl 9'-(2-cyclopropylethoxy)-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (50 mg, 0.12 mmol) in MeOH (2 ml_) and water (0.5 ml_) was added LiOH (1 .2 mg, 0.05 mmol) at room temperature The reaction mixture was stirred at room temperature for 1 hand acidified to pH~4 by addition of 1 N HCI. The reaction mixture was extracted with 15% IPA in DCM twice and the combined organic layers were dried over Na2S04, filtered and concentrated. The residue was purified by reverse phase HPLC (C18, 0 - 40%, MeCN in H20 with 0.1 % formic acid) to afford the title compound (22 mg, 48% yield) as a white solid. LCMS (ESI) m/z: 380.8 (M + 1)+.1H NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1 H), 7.86 (s, 1 H), 7.25 (s, 1 H), 7.05 (s, 1 H), 4.14 (t, J = 6.4 Hz, 2H), 3.32 (s, 2H), 2.67 - 2.59 (m, 2H), 2.20 (s, 3H), 2.06 - 1 .99 (m, 2H), 1 .94 - 1 .85 (m, 2H), 1 .72 - 1 .66 (m, 2H), 0.90 - 0.84 (m, 1 H), 0.49 - 0.43 (m, 2H), 0.19 - 0.13 (m, 2H).
[001041] Example 81
[001042] 10'-Methyl-2'-oxo-9'-(pyrrolidin-1 -yl)-2',7'-dihvdrospiro[cvclobutane-
Figure imgf000216_0002
[001043] Step 1 :Ethyl 10'-methyl-2'-oxo-9'-(((trifluoromethvnsulfonyl)oxyV2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000217_0001
[001044] To a solution of ethyl 9'-hydroxy-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (2 g, 5.9 mmol) in CH3CN was added PhN(Tf)2 (2.5 g, 7.1 mmol), DIPEA (2.3 g, 17.7 mmol), and DMAP (36 mg, 0.295 mmol). The reaction mixture was stirred at room temperature for 2 h. Then the mixture was filtered, and the solid was washed with EtOAc. The filtrate was evaporated and the residue was purified by chromatography (silica gel, Ethyl acetate/iso-hexane, 1 : 19) to afford the title compound (1 .3 g, 47% yield). LCMS (ESI) m/z: 472.2 (M + 1)+.
[001045] Step 2: Ethyl 9'-(((cvclopropylmethyl)(methyl)amino-10'-methyl-2'-
Figure imgf000217_0002
[001046] A Schlenk tube was charged with Pd(dba)2 (120.75 mg, 0.2 mmol), BINAP (130.76 mg, 0.21 mmol), Cs2C03 (137 mg, 0.42 mmol) and toluene (2 ml_). To this mixture was added a solution of ethyl 10'-methyl-2'-oxo-9'-(((trifluoromethyl)sulfonyl)oxy)- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.21 mmol) and 1 -cyclopropyl-N-methylmethanamine44 mg, 0.63 mmol) in toluene (2 ml_) via syringe. The mixture was heated to 100°C with stirring until the title compound had been consumed as judged by LCMS analysis (2 - 8 h). The mixture was cooled to room temperature, taken up in ethyl acetate, filtered, and concentrated. The crude material was purified by silica gel chromatography to afford the title compound (59 mg, 72% yield). LCMS (ESI) m/z: 407.4 (M + 1)+.
[001047] Step 3: 9'-((Cvclopropylmethyl)(methyl)amino)-10'-methyl-2'-oxo- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000217_0003
[001048] To a solution of ethyl 9'-((cyclopropylmethyl)(methyl)amino)-10'- methyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate(20 mg, 0.05 mmol) in MeOH (2 mL) and water (0.5 mL) was added LiOH (1 .2 mg, 0.05 mmol) at room temperature The reaction mixture was stirred at room temperature for 1 h, and detected with LCMS. After the completion of the reaction, the mixture was purified by reverse phase HPLC (C18 0-30% MeCN in H20 with 0.1 % formic acid) to afford the title compound (9 mg, 50% yield) as a yellow solid. LCMS (ESI) m/z: 379.4 (M + 1 )+. 1H NMR (400 MHz, DMSO-d6) δ 8.75(s, 1 H), 7.68(s, 1 H), 7.15(s, 1 H), 6.70 (s, 1 H), 3.41 - 3.38 (m, 4H), 3.24(s, 2H), 2.64 - 2.57 (m, 2H), 2.38 (s, 3H), 2.04 - 2.00 (m, 2H), 1 .93 - 1 .88 (m, 6H).
[001049] Example 82
[001050] 9'-lsobutoxy-10'-methyl-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'- pyrido[2,1 -alisoquinolin -3'-carboxylic acid
Figure imgf000218_0001
[001051] Step 1 : Ethyl 9'-isobutoxy-10'-methyl^'-oxo^'J1- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000218_0002
[001052] To the solution of ethyl 9'-hydroxy-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (90 mg, 0.265 mmol) in DMF (5 mL) was added K2C03 (73.2 mg, 0.530 mmol) and 1 -bromo-2-methylpropane (43.6mg, 0.318 mmol), and the resulting mixture was heated to 90°C for 3 hours. After cooling to room temperature, the reaction mixture was partitioned between EtOAc and H20, and the organic layer was separated. The aqueous layer was extracted with EtOAc twice. The combined organic layers were washed with brine, dried over Na2S04, filtered and concentrated. The residue was purified by chromatography (silica gel, 0 - 20% MeOH in DCM) to afford the title compound. (60 mg, 57.2% yield). LCMS (ESI) m/z: 396.3 (M + 1 )+.
[001053] Step 2: g'-lsobutoxy-I O'-methyl^'-oxo^'J1- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000219_0001
[001054] Sodium hydroxide (48.5 mg, 1 .214mmol) dissolved in H20 (1 ml) was added to a solution of ethyl 9'-isobutoxy-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (60 mg, 0.151 mmol) in MeOH (5 ml). The resulting mixture was stirred at room temperature for 2 h and then acidified to ph~5 with 1 N HCI. The mixture was diluted with EtOAc (20 mL) and H20 (30 ml_), and the aqueous layer was extracted with EtOAc (50 ml_x2). The combined organic phase was dried over anhydrous Na2S04, filtered, and concentrated. The residue was purified by Gilson (C18 column, 30%-100%MeCN in H20,with 0.1 % formic acid in H20) to provide the title compound (10 mg, 17.9% yield) as a white solid. LCMS (ESI) m/z: 368.3 (M + 1 )+. Ή NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1 H), 7.86 (s, 1 H), 7.25 (s, 1 H), 7.03 (s, 1 H), 3.87 (d, J = 6.4 Hz, 2H), 3.34 (s, 2H), 2.68 - 2.60 (m, 2H), 2.21 (s, 3H), 2.14 - 2.08 (m, 1 H), 2.04 - 1 .99 (m, 2H), 1 .93 - 1 .86 (m, 2H), 1 .02 (d, J = 6.7 Hz, 6H).
[001055] Example 83
[001056] 9'-(2-Hvdroxy-2-methylpropoxy)-10'-methyl-2'-oxo-2'.7'- dihvdrospiro[cvclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000219_0002
[001057] Step 1 : Ethyl 9'-(2-hvdroxy-2-methylpropoxy)-10'-methyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000219_0003
[001058] To a solution of ethyl 9'-hydroxy-10'-methyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (200 mg, 0.59 mmol) in DMF (3 mL) was added K2C03 (407.1 mg, 2.95 mmol), Nal (353.7 mg, 2.36 mmol), and 1 -chloro-2-methylpropan-2-ol (256.2 mg, 2.36 mmol). The reaction mixture was stirred at 90 °C for 2 h. Then the reaction was extracted with DCM twice, the combined organic phases were washed with brine, dried over Na2S04, filtered and concentrated. The residue was purified by chromatography (silica gel, 0-40% CH3OH in DCM) to afford the title compound (106 mg, 43.7% yield). LCMS (ESI) m/z: 412.4 (M + 1)+.
[001059] Step 2: 9'-(2-Hvdroxy-2-methylpropoxyV10'-methyl-2'-oxo-2'.7'- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000220_0001
[001060] To a solution of ethyl 9'-(2-hydroxy-2-methylpropoxy)-10'-methyl-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (46.6 mg, 0.1 1 mmol) in MeOH (3ml_) was added NaOH (13.6mg, 0.34 mmol) in water (1 mL). The resulting mixture was stirred at room temperature for 30 minutes. After acidified to pH 6 by addition of 1 N HCI, the reaction mixture was extracted with 15% IPA in DCM twice, the combined organic phases were dried over Na2S04, filtered, concentrated, and the residue was purified by reverse phase HPLC (Gilson, Ci8 30%-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound as a white solid (17 mg, 39% yield). LCMS (ESI) m/z: 384.8 (M + 1)+. Ή NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1 H), 7.86 (s, 1 H), 7.25 (s, 1 H), 7.02 (s, 1 H), 4.70 (s, 1 H), 3.82 (s, 2H), 3.32 (s, 2H), 2.68 - 2.60 (m, 2H), 2.23 (s, 3H), 2.05 - 1 .99 (m, 2H), 1 .93 - 1 .85 (m, 2H), 1 .25 (s, 6H).
[001061] Example 84
[001062] 9'-(2-methoxy-2-methylpropoxy)-10'-methyl-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000220_0002
[001063] Sodium hydride (35.04 mg, 60% in mineral oil, 1 .46 mmol) was added portion-wise to a solution of ethyl 9'-(2-hydroxy-2-methylpropoxy)-10'-methyl-2'-oxo- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate(60 mg, 0.15 mmol) in anhydrous THF (5 mL) at 0 °C. After 15 minutes, CH3I (41 .44 mg, 0.30 mmol) was added and the resulting mixture was warmed to room temperature and stirred for 1 h. The reaction mixture was poured into ice water. LC-MS indicated the ester was hydrolyzed, then the above mixture was acidified with 1 N HCI to PH 6, extracted with 15% IPA in DCM twice. The combined organic phases were dried over Na2S04, filtered, concentrated, and the residue was purified by reverse phase HPLC (Gilson, C18, 30%-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound as a white solid (13 mg, 22% yield). LCMS (ESI) m/z: 398.3 (M + 1)M H NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1 H), 7.87 (s, 1 H), 7.26 (s, 1 H), 7.06 (s, 1 H), 3.96 (s, 2H), 3.32 (s, 2H), 3.20 (s, 3H),2.68 - 2.59 (m, 2H), 2.22 (s, 3H), 2.06 - 1.98 (m, 2H), 1.94 - 1.85 (m, 2H), 1.26 (s, 6H).
[001064] Example 85
[001065] g'^Furan-S-vD^'-oxo-I O'^pyrrolidin-l-vD^'J1- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000221_0001
dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000221_0002
[001067] A mixture of ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (275mg, 0.56 mmol), Cs2C03 (1.1 g, 3.36 mmol), BINAP ( 52.3mg, 0.084mmol), Pd2(dba)3 ( 51 .28 mg, 0.056 mmol), and pyrrolidine (insert text) in toluene (5 mL) was stirred at 100°C for 1.5 hours. The solvent was removed by evaporation and the residue was purified by chromatography (silica gel, 0-70% EtOAc in petroleum ether) to afford the title compound (200 mg, 74.3% yield), LCMS (ESI) m/z: 485.4 (M + 1)+.
[001068] Step 2:
Figure imgf000221_0003
dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000221_0004
[001069] To a solution of ethyl 9'-(benzyloxy)-2'-oxo-10'-(pyrrolidin-1 -yl)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (200 mg, 0.413 mmol) in CH3OH (5 ml_) was added Pd/C (100 mg). The reaction mixture was stirred at room temperature under H2 atmosphere (~1 atm) overnight, filtered and concentrated. The residue was used directly in the next step, (assumed quantitative). LCMS (ESI) m/z: 395.4 (M + 1)+.
[001070] Step 3: Ethyl 2'-oxo-10'-f pyrrolidin-1 -vD-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel- 3'-carboxylate
Figure imgf000222_0001
[001071] To a solution of ethyl 9'-hydroxy-2'-oxo-10'-(pyrrolidin-1 -yl)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (200 mg, 0.51 mmol) in DCM (5 mL) was added PhNTf2 (271 .7 mg, 0.76 mmol), DMAP (6.2 mg, 0.05 mmol) and TEA (102.6 mg, 1.01 mmol). The resulting mixture was stirred at room temperature for 2 hours. The solvent was removed by evaporation and the residue was purified by chromatography (silica gel, 0-30% MeOH in DCM) to afford the title compound (148 mg, 57.8% yield), LCMS (ESI) m/z: 527.2 (M + 1)+.
[001072] Step 4: Ethyl g'^furan-S-vD^'-oxo-I O'^pyrrolidin-l-vD^'J'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000222_0002
[001073] To a solution of ethyl 2'-oxo-10'-(pyrrolidin-1 -yl)-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate (148 mg, 0.28 mmol) in toluene (5 mL) and water (1 drop) was added Pd(PPh3)4 (34.5 mg, 0.028 mmol), Cs2C03 (461 .54 mg, 1.41 mmol) and 3-furanboronic acid (62.88 mg, 0.56 mmol). The resulting mixture was stirred at 100°C for 2 hours. The solvent was removed by evaporation and the residue was purified by chromatography (silica gel, 0- 30% MeOH in DCM) to afford the title compound (90 mg, 72% yield). LCMS (ESI) m/z: 445.4 (M + 1)+. [001074] Step 5: 9'-(Furan-3-yl)-2'-oxo-10'-(pyrrolidin-1 -νΠ-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000223_0001
[001075] To a solution of ethyl 9'-(furan-3-yl)-2'-oxo-10'-(pyrrolidin-1 -yl)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (90 mg, 0.20 mmol) in THF (4ml_) was added LiOH (19.4 mg, 0.81 mmol) in H20 (1 mL). The resulting mixture was stirred at room temperature for 1 h. After acidified by addition of 1 N HCI to pH 6, the reaction mixture was extracted with 15% IPA in DCM twice. The combined organic phases were dried over Na2S04, filtered, concentrated, and the residue was purified by reverse phase HPLC (Gilson, Ci8, 30%-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound as a yellow solid (24 mg, 28.5% yield). LCMS (ESI) m/z: 417.8 (M + 1)+. Ή NMR (400 MHz, DMSO-de) δ 8.83 (s, 1 H), 8.00 (s, 1 H), 7.76 (t, J = 1.6 Hz, 1 H), 7.45 (s, 1 H), 7.43 (s, 1 H), 7.38 (s, 1 H), 6.85 (d, J = 1.0 Hz, 1 H) , 3.27 (s, 2H), 3.08 - 3.02 (m, 4H), 2.68 - 2.61 (m, 2H), 2.06 - 2.00 (m, 2H), 1.94 - 1 .86 (m, 2H), 1.85 - 1.79 (m, 4H).
[001076] Example 86
[001077] 3-(benzyloxy)-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000223_0002
[001078] Step 1 : 3-(Benzyloxy)-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000223_0003
[001079] To a solution of ethyl 3-(benzyloxy)-10'-methoxy-9'-(3- methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'- carboxylate (100 mg, 0.187 mmol) in THF/EtOH/H20 (2 mL /1 mL /0.5 mL) was added LiOH (31.5 mg, 0.75 mmol). The reaction mixture was stirred at r.t for 1 hr and then neutralized with 1 N HCI and extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (68 mg, 71.8% yield ) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 10.15 (br, 1 H), 8.75 (s, 1 H), 7.49 (s, 1 H), 7.45 (s, 1 H), 7.38 - 7.27(m, 5H),7.01 (s, 1 H),4.42 (s,2 H), 4.31 - 4.25 (m, 1 H), 4.11 (t, J = 6.4 Hz, 2H), 3.88 (s, 3H), 3.48 (t, J = 6.2 Hz, 2H), 3.33 (s, 2H), 3.25 (s, 3H),2.88 (dd, J = 13.7, 7.3 Hz, 2H), 2.22 (dd, J = 13.9,4.6 Hz, 2H), 2.03-1.96 (m, 2H). LCMS (ESI) m/z: 506.3 (M + 1)+.
[001080] Example 87
[001081] 10'-methoxy-2'-oxo-9'-pentyl-2',7'-dihvdrospiro[cvclobutane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000224_0001
[001083] 1-Pentyne (47 mg, 0.41 mmol) was added to a mixture of ethyl 10'- methoxy-2'-oxo-9'-(((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylate (150 mg, 0.317 mmol) , Pd(PPh3)2Cl2 (44.5 mg,0.063mmol), K2C03 (87.5 mg,0.63mmol) , Cul (2.4 mg, 0.013mmol) in DMF (5 ml),. The resulting mixture was stirred at 60 °C for 3 h, diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50ml_x2). The combined organic phase was washed with brine, dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography(silica gel,0-10%MeOH in DCM) to afford ethyl 10'-methoxy-2'-oxo-9'-(pent- 1-yn-1-yl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (120 mg, 93.3% yield) as a yellow solid. LCMS (ESI) m/z: 406.3 (M + 1)+.
[001084] Step 2: Ethyl 10'-methoxy-2'-oxo-9'-pentyl-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000225_0001
[001085] A mixture of 10'-methoxy-2'-oxo-9'-(pent-1-yn-1 -yl)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (120 mg, 0.296 mmol) and 10% wet Pd/C (100 mg) in MeOH (10 ml_) was stirred under hydrogen atmosphere (15 psi) at r. t. for 1 h.The mixture was filtered and concentrated under reduced pressure to afford ethyl 10'-methoxy-2'-oxo-9'-pentyl-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 82.5% yield) as a yellow solid. LCMS (ESI) m/z: 410.3 (M + 1)+.
[001086] Step 3: 10'-Methoxy-2'-oxo-9'-pentyl-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000225_0002
[001087] To a solution of ethyl 10'-methoxy-2'-oxo-9'-pentyl-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.244 mmol) in MeOH (10 ml), was added a solution of NaOH (78.1 mg, 1.952mmol) in H20 (1 ml). The resulting mixture was stirred at room temperature for 2 h and then acidified to ph~5 with 1 N HCI. The mixture was diluted with EtOAc (20 mL) and H20 (30 ml_), and then extracted with EtOAc (50 ml_x2). The combined organic phase was dried over anhydrous Na2S04 and concentrated. The residue was purified by Gilson (C18 column, 50%-100%MeCN in H20, with 0.1 % formic acid in H20) to give 10'-methoxy-2'-oxo-9'-pentyl-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylic acid (29.3 mg, 31 .5% yield) as a white solid. LCMS (ESI) m/z: 382.5 (M + 1)+. Ή NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1 H), 7.52 (s, 1 H), 7.47 (s, 1 H), 7.23 (s, 1 H), 3.91 (s, 3H), 3.27 (s, 2H), 2.67 - 2.57 (m, 4H), 2.02 - 1 .97 (m, 2H), 1.95 - 1.85 (m, 2H), 1.60 - 1.53 (m, 2H), 1.34 - 1 .28 (m, 4H), 0.87 (t, J = 7.0 Hz, 3H).
[001088] Example 88
[001089] 10'-Methoxy-9'-(4-methoxyphenvn-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000226_0001
[001090] Step 1 : Ethyl 10'-methoxy-9'-(4-methoxyphenyl)-2'-oxo-2'.7'- dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000226_0002
[001091] A mixture of ethyl 10'-methoxy-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate (80 mg,0.164mmol), (4-methoxyphenyl)boronic acid (37.4 mg, 0.246 mmol), Pd(dppf)CI2 (13.4 mg, 0.0164 mmol), CH3COOK (48 mg, 0.492 mmol) in 1 ,4-dioxane (1 mL) and H2O (1 drop) was stirred at 80°C under N2 atmosphere for 3 h. The mixture was concentrated and the residue was purified by chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (40 mg, 54.8% yield) as a yellow solid. LCMS (ESI) m/z: 446.4(M + 1)+.
[001092] Step 2: 10'-Methoxy-9'-(4-methoxyphenyl)-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000226_0003
[001093] To a solution of ethyl 10'-methoxy-9'-(4-methoxyphenyl)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (40 mg, 0.09 mmol) in THF/EtOH/H20 (2 mL /1 mL /0.5 mL) was added LiOH (23 mg, 0.54 mmol). After stirring at r.t for 1 hr, the resulting mixture was neutralized with 1 N HCI and extracted with DCM/i- PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0- 100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (26.5 mg, 70.7% yield ) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 10.15 (br, 1 H), 8.85 (s, 1 H), 7.61 (d, J = 3.6 Hz, 2H), 7.52 (d, J = 8.6 Hz, 2H), 7.40 (s, 1 H), 7.01 (d, J = 8.6 Hz, 2H), 3.90 (s, 3H), 3.81 (s, 3H), 3.34 (s,2H), 2.70-2.63 (m,2H),2.09-2.02 (m, 2H), 1.91 -1.85 (m, 2H). LCMS (ESI) m/z: 418.32(M + 1)+.
[001094] Example 89
[001095] 9l-(4-Cvanophenyl)-10l-methoxy-2'-oxo-2l.7l- dihvdrospiro[cvclobutan -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000227_0001
[001096] Step 1 : Ethyl 9'-(4-cvanophenylV10'-methoxy-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000227_0002
[001097] A mixture of ethyl 10'-methoxy-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate (80mg,0.164mmol), (4-cyanophenyl)boronic acid (36 mg, 0.246 mmol), Pd(dppf)CI2 (13.4 mg, 0.0164mmol), CH3COOK (48 mg, 0.492 mmol) in 1 ,4-dioxane (1 mL) and H20 (1 drop) was stirred at 80°C under N2 atmosphere for 3 h. The mixture was concentrated and the residue was purified by chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (50 mg, 69.4% yield) as a yellow solid. LCMS (ESI) m/z: 441 .3(M + 1)+.
[001098] Step 2: 9'-(4-Cvanophenyl)-10'-methoxy-2'-oxo-2'.7'- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000227_0003
[001099] To a solution of ethyl 10'-methoxy-9'-(4- cyanophenyl)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate(50 mg, 0.113 mmol) in THF/EtOH/H20(2 mL /1 mL /0.5 mL) was added LiOH (28.6 mg, 0.68 mmol). After stirring at r.t for 1 hr, the resulting mixture was neutralized with 1 N HCI and extracted with DCM/i- PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0- 100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (16 mg, 34.2% yield ) as a white powder. 1H NMR (400 MHz, DMSO-de) δ 10.15 (br, 1 H), 8.86 (s, 1 H), 7.93 (d, J = 7.9 Hz, 2H), 7.78 (d, J = 7.9 Hz, 2H),7.68 (d, J = 11 .5 Hz, 2H), 7.49 (s, 1 H), 3.93 (s, 3H),3.36 (s, 2H),2.71 - 2.64 (m, 2H), 2.08 - 2.02 (m, 2H),1 .94 - 1 .87 (m, 2H). LCMS (ESI) m/z: 413.3 (M + 1)+.
[001100] Example 90
[001101] 10l-Methoxy-9l-(2-methoxyphenvn-2'-oxo-2l.7l- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000228_0001
[001102] Stepl : Ethyl 10'-methoxy-9'-(2-methoxyphenyr)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000228_0002
[001103] A mixture of ethyl 10'-methoxy-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate (80mg, 0.164mmol), (2-methoxyphenyl)boronic acid (37.4 mg, 0.246 mmol), Pd(dppf)CI2 (13.4 mg, 0.0164mmol), CH3COOK (48 mg, 0.492 mmol) in 1 ,4-dioxane (1 mL) and H20 (1 drop) was stirred at 80°C under N2 atmosphere overnight. The mixture was concentrated and the residue was purified by chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (40 mg, 54.8% yield) as a yellow solid. LCMS (ESI) m/z: 446.4 (M + 1)+.
[001104] Step 2: 10'-Methoxy-9'-(2-methoxyphenyl)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000229_0001
[001105] To a solution of ethyl 10'-methoxy-9'-(2-methoxyphenyl)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate(40 mg, 0.09 mmol) in THF/EtOH/H20 (2 mL /1 mL /0.5 mL) was added LiOH (23 mg, 0.54 mmol). After stirring at r.t for 1 hr, the resulting mixture was acidified with 1 N HCI and extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (26.5 mg, 70.7% yield ) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 15.94 (br, 1 H), 8.96 (s, 1 H), 7.39 (dd, J =11.3, 4.4 Hz, 1 H), 7.26 - 7.20 (m, 4H), 7.03 (dd, J =16.1 , 8.0 Hz, 2H), 3.85 (s, 3H), 3.80(s, 3H), 3.24(s, 2H), 2.69 - 2.61 (m, 2H), 2.25 - 2.18 (m, 2H), 2.07 - 1.97 (m, 2H). LCMS (ESI) m/z: 418.3 (M + 1)+.
[001106] Example 91
[001107] g'-O-CvanophenvD-I O'-methoxy^'-oxo^'J1- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000229_0002
[001109] A mixture of ethyl 10'-methoxy-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate (80mg,0.164mmol), (3-cyanophenyl)boronic acid (36 mg, 0.246 mmol), Pd(dppf)CI2 (13.4 mg, 0.0164mmol), CH3COOK (48 mg, 0.492 mmol) in 1 ,4-dioxane (1 mL) and H20 (1 drop) was stirred at 80°C under N2 atmosphere for 3 h. The mixture was concentrated and the residue was purified by chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (38 mg, 52.8 % yield) as a yellow solid. LCMS (ESI) m/z: 441 .3 (M + 1)+.
[001110] Step 2: 9'-(3-Cvanophenyl)-10'-methoxy-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000230_0001
[001111] To a solution of ethyl 10'-methoxy-9'-(3- cyanophenyl)-2'-oxo-2',7'- dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (38 mg, 0.086 mmol) in THF/EtOH/H20 (2 mL /1 mL /0.5 mL) was added LiOH (22 mg, 0.518 mmol). After stirring at r.t for 1 hr, the resulting mixture was acidified with 1 N HCI and extracted with DCM/i- PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0- 100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (20 mg, 56.2% yield ) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 10.25 (br, 1 H), 8.86 (s, 1 H), 8.04-8.02 (m,1 H),7.93 (d, J = 8.1 Hz, 1 H), 7.86 (d, J = 7.8 Hz, 1 H), 7.69 (s, 1 H), 7.68-7.64 (m, 2H), 7.51 (s, 1 H), 3.94 (s, 3H), 3.35 (s, 2H), 2.71 -2.65 (m, 2H), 2.08-2.02 (m, 2H), 1.93-1.87 (m, 2H). LCMS (ESI) m/z: 413.3 (M + 1)+.
[001112] Example 92
[001113] I Q'-Methoxy-g'-O-methoxyphenvD^'-oxo^'J'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000230_0002
[001114] Stepl : Ethyl 10'-methoxy-9'-(3-methoxyphenyl)-2'-oxo-2'.7' dihvdro spiro [cyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000231_0001
[001115] A mixture of ethyl 10'-methoxy-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate (90mg, 0.185mmol), (3-methoxyphenyl)boronic acid (42 mg, 0.276 mmol), Pd(dppf)CI2 (15 mg, 0.0185mmol), CH3COOK (55 mg, 0.55 mmol) in 1 ,4-dioxane (1 mL) and H2O (1 drop) was stirred at 80°C under N2 atmosphere for 6 h. The mixture was concentrated and the residue was purified by chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (44 mg, 53.6% yield) as a yellow solid. LCMS (ESI) m/z: 446.4 (M + 1)+.
[001116] Step 2: 10'-Methoxy-9'-(3-methoxyphenyl)-2'-oxo-2',7'-dihydrospiro [cvclobutane-1 ,6'-pyrid -alisoquinolinel-3'-carboxylic acid
Figure imgf000231_0002
[001117] To a solution of ethyl 10'-methoxy-9'-(3-methoxyphenyl)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (44 mg, 0.1 mmol) in THF/EtOH/H20 (2 mL /1 mL /0.5 mL) was added LiOH (25 mg, 0.6 mmol). After stirring at r.t for 1 hr, the resulting mixture was acidified with 1 N HCI and extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (28.5 mg, 69.2% yield ) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 11.94 (br, 1 H), 8.85 (s, 1 H), 7.63 (d, J =5.4 Hz, 2H), 7.43 (s, 1 H), 7.36 (t, J =7.9 Hz, 1 H), 7.15-7.08 (m, 2H), 6.96 (dd, J = 8.1 ,1.9 Hz, 1 H), 3.91 (s, 3H), 3.80 (s, 3H), 3.35 (s, 2H), 2.71 -2.63 (m, 2H), 2.09 - 2.01 (m, 2H),1 .96-1.86 (m, 2H). LCMS (ESI) m/z: 418.3 (M + 1)+.
[001118] Example 93 [001119] 9l-(2-CvanophenylV10l-methoxy-2'-oxo-2l.7l- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000232_0001
[001120] Step 1 : Ethyl9'-(2-cvanophenyl)-10'-methoxy-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000232_0002
[001121] A mixture of ethyh 0'-methoxy-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate(90mg,0.185mmol), (2-cyanophenyl)boronic acid (41 mg, 0.276 mmol), Pd(dppf)CI2 (15 mg, 0.0185mmol), CH3COOK (55 mg, 0.55 mmol) in 1 ,4-dioxane(1 mL) and H20(1 drop) was stirred at 80°C under N2 atmosphere for 6 h. The mixture was concentrated, and the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (40 mg, 49.4% yield) as a yellow solid. LCMS (ESI) m/z: 441 .3 (M + 1)+.
[001122] Step 2: 9'-(2-Cvanophenyl)-10'-methoxy-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000232_0003
[001123] To a solution of ethyH 0'-methoxy-9'-(2- cyanophenyl)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 , 6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate(40 mg, 0.09 mmol) in THF/EtOH/H20 (2 mL /1 mL /0.5 mL) was added LiOH (23 mg, 0.54 mmol). After stirred at r.t for 1 hr, the resulting mixture was neutralized with 1 N HCI and extracted with DCM/i- PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0- 100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (13.2 mg, 35.2% yield ) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 10.25 (br, 1 H), 8.87 (s, 1 H), 7.94 (d, J = 7.1 Hz, 1 H), 7.82 - 7.78 (m, 1 H), 7.73 (s, 1 H), 7.69 (s, 1 H), 7.63 - 7.57 (m, 2H), 7.42 (s, 1 H), 3.91 (s, 3H), 3.36 (s, 2H), 2.72 - 2.65 (m, 2H), 2.08 - 2.01 (m, 2H), 1.95 - 1 .87 (m, 2H). LCMS (ESI) m/z: 413.3 (M + 1)+.
[001124] Example 94
[001125] 10'-Cethoxy-2'-oxo-9'-(pyridin-4-yl)-2',7'-dihvdrospiro[cyclobutane-
Figure imgf000233_0001
[001126] Step 1 : Ethyl I Q'-methoxy^'-oxo-g'-foyridin^-vn^'J'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000233_0002
[001127] A mixture of ethyll 0'-methoxy-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate(70 mg,0.14 mmol), pyridin-4-ylboronic acid (32 mg, 0.26 mmol), Pd(dppf)CI2 (11.4 mg, 0.014mmol), CH3COOK (42 mg, 0.42 mmol) in 1 ,4-dioxane(1 mL) and H20(1 drop) was stirred at 80°C under N2 atmosphere overnight. The mixture was concentrated, the residue was purified by flash chromatography (silica gel, 0 - 20% MeOH in DCM) to afford the title compound (60 mg, 100% yield) as a yellow solid. LCMS (ESI) m/z: 417.3 (M + 1)+.
[001128] Step 2: I Q'-Methoxy^'-oxo-g'-foyridin^-vn^'J'- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000233_0003
[001129] To a solution of ethyl 10'-methoxy-2'-oxo-9'-(pyridin-4-yl)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (60 mg, 0.144 mmol) in THF/EtOH/H20(2 mL /1 mL /0.5 mL) was added LiOH (36 mg, 0.86 mmol). After stirring at r.t for 1 hr, the resulting mixture was neutralized with 1 N HCI, then basified with NH3/H20 , extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (35.5 mg, 63.4% yield ) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 11.94 (br, 1 H), 8.86 (s, 1 H), 8.65 (d, J =5.8 Hz, 2H), 7.70 (s, 1 H), 7.67(s, 1 H), 7.59 (d, J =5.8 Hz, 2 H), 7.53 (s, 1 H), 3.95 (s, 3H), 3.36 (s, 2H), 2.71 - 2.65 (m, 2H), 2.07 - 2.02 (m, 2H), 1 .94 - 1 .87 (m, 2H). LCMS (ESI) m/z: 389.3 (M + 1)+.
[001130] Example 95
[001131] 10l-Methoxy-9l-(2-methoxypyridin-3-vn-2'-oxo-2l.7l- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000234_0001
[001132] Step 1 : Ethyl10'-methoxy-9'-(2-methoxypyridin-3-yl)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000234_0002
[001133] A mixture of ethyll 0'-methoxy-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate(70 mg,0.14 mmol), pyridin-4-ylboronic acid (40 mg, 0.26 mmol), Pd(dppf)CI2 (11.4 mg, 0.014mmol), CH3COOK (42 mg, 0.42 mmol) in 1 ,4-dioxane(1 mL) and H20(1 drop) was stirred at 80°C under N2 atmosphere overnight. The mixture was concentrated, the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (57 mg, 89% yield) as a yellow solid. LCMS (ESI) m/z: 447.3 (M + 1)+.
[001134] Step 2: 10'-Methoxy-9'-(2-methoxypyridin-3-vn-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000235_0001
[001135] To a solution of ethyl10'-methoxy-9'-(2-methoxypyridin-3-yl)-2'-oxo- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (57 mg, 0.128 mmol) in THF/EtOH/H20 (2 mL /1 mL /0.5 mL) was added LiOH (32 mg, 0.767 mmol). After stirred at r.t for 1 hr, the resulting mixture was neutralized with 1 N HCI, then basified with NH3/H2O, extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (30 mg, 56.2% yield ) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 11 .94 (br, 1 H), 8.86 (s, 1 H), 8.22 - 8.19 (m, 1 H), 7.67 - 7.61 (m, 3H),7.31 (s, 1 H), 7.08 (dd, J =7.0, 5.1 Hz,1 H), 3.86 (s, 3H), 3.82 (s, 3H), 3.32 (s, 2H), 2.72 - 2.64 (m, 2H), 2.08 - 2.01 (m, 2H), 1.94 - 1.86 (m, 2H). LCMS (ESI) m/z: 419.3 (M + 1)+.
[001136] Example 96
[001137] 9l-(2-Cvclopropylethyl)-10l-methoxy-2'-oxo-2l.7l- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000235_0002
[001138] Step 1 : Ethyl9'-(cvclopropylethvnyl)-10'-methoxy-2'-oxo- 2',7'dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000235_0003
[001139] A mixture of ethyll 0'-methoxy-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate(85mg,0.174 mmol), ethynylcyclopropane (23 mg, 0.35 mmol), Pd(PPh3)2CI2 (24 mg, 0.035 mmol), Cul(6.6 mL, 0.035 mmol) and TEA (0.1 mL, 0.696 mmol) in DMF(2 mL) was stirred at 90°C under N2 atmosphere overnight. The mixture was concentrated, the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (30 mg, 43% yield) as a yellow solid. LCMS (ESI) m/z: 404.3 (M + 1)+.
[001140] Step 2: Ethyl9'-(2-cvclopropylethyl)-10'-methoxy-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000236_0001
[001141] To a solution of ethyl 9'-(cyclopropylethynyl)-10'-methoxy-2'-oxo- 2',7'-dihydrospiro [cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate(30 mg, 0.074 mmol) in EtOH (5 mL) was added palladium (10 mg, 5% on carbon (wetted with ca. 55% Water)), the resulting mixture was stirred at r.t under H2 overnight. Palladium was removed by filtering , the filtrate was concentrated to afford the title compound (28 mg, 92% yield) as a yellow solid. LCMS (ESI) m/z: 408.3 (M + 1)+.
[001142] Step 3: 9'-(2-Cvclopropylethyl)-10'-methoxy-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000236_0002
[001143] To a solution of ethyl 9'-(2-cyclopropylethyl)-10'-methoxy-2'-oxo-2',7'- dihydro spiro [cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (28 mg, 0.069 mmol) in THF/EtOH/H20(2 mL /1 mL /0.5 mL) was added LiOH (17 mg, 0.412 mmol). After stirred at r.t for 1 hr, the resulting mixture was neutralized with 1 N HCI, extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0- 100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (9 mg, 34.6% yield ) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 1 1.94 (br, 1 H), 8.82 (s, 1 H), 7.52 (s, 1 H), 7.47 (s, 1 H), 7.24 (s, 1 H), 3.91 (s, 3H), 3.26 (s, 2H), 2.72 - 2.61 (m, 4H), 2.03 - 1.97 (m, 2H), 1.94 - 1.85 (m, 2H),1 .49 - 1.43 (m, 2H) 0.74 - 0.69 (m, 1 H) 0.42 - 0.37 (m, 2H) 0.04 - 0.01 (m, 2H). LCMS (ESI) m/z: 380.2 (M + 1)+.
[001144] Example 97 [001145] 10'-Methoxy-9'-(6-methoxypyridin-3-yl)-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000237_0001
[001146] Step 1 : Ethyl10'-methoxy-9'-(6-methoxypyridin-3-ylV2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000237_0002
[001147] A mixture of ethyll 0'-methoxy-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate(80mg, 0.164 mmol), (6-methoxypyridin-3-yl)boronic acid (45 mg, 0.295 mmol), Pd(dppf)CI2 (13mg, 0.0164mmol), CH3COOK (48 mg, 0.492 mmol) in 1 ,4-dioxane(1 mL) and H20 (1 drop) was stirred at 80°C under N2 atmosphere overnight. The mixture was concentrated, the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (61 mg, 83.5% yield) as a yellow solid. LCMS (ESI) m/z: 447.3 (M + 1)+.
[001148] Step 2: 10'-Methoxy-9'-(6-methoxypyridin-3-vn-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000237_0003
[001149] To a solution of ethyll 0'-methoxy-9'-(6-methoxypyridin-3-yl)-2'-oxo- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (61 mg, 0.137 mmol) in THF/EtOH/H20(2 mL /1 mL /0.5 mL) was added LiOH (34 mg, 0.82 mmol). After stirred at r.t for 1 hr, the resulting mixture was neutralized with 1 N HCI, then basified with NH3/H20, extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (24 mg, 42.1 % yield ) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 11 .94 (br, 1 H), 8.86 (s, 1 H), 8.37 (d, J = 2.1 Hz ,1 H), 7.92 (dd, J = 8.6, 2.3 Hz,1 H),7.65 (s, 1 H), 7.63 (s, 1 H), 7.46 (s, 1 H), 6.91 (d, J =8.6 Hz,1 H), 3.93 (s, 3H), 3.90 (s, 3H), 3.34 (s, 2H), 2.71 - 2.64 (m, 2H), 2.09 - 2.03 (m, 2H), 1.94 - 1.85 (m, 2H). LCMS (ESI) m/z: 419.2 (M + D+.
[001150] Example 98
[001151] 10'-Methoxy-9'-(5-methoxypyridin-3-vn-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000238_0001
[001152] Step 1 : Ethyl10'-methoxy-9'-(5-methoxypyridin-3-yl)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000238_0002
[001153] A mixture of ethyll 0'-methoxy-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate(100mg,0.205mmol), (5-methoxypyridin-3-yl)boronic acid (56 mg, 0.369 mmol), Pd(dppf)CI2 (17 mg, 0.021 mmol), CH3COOK (60 mg, 0.615 mmol) in 1 ,4-dioxane(1 mL) and H20(1 drop) was stirred at 80°C under N2 atmosphere overnight. The mixture was concentrated, the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (70 mg, 76.4% yield) as a yellow solid. LCMS (ESI) m/z: 447.3 (M + 1)+.
[001154] Step 2:10'-Methoxy-9'-(5-methoxypyridin-3-yl)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000239_0001
[001155] To a solution of ethyl10'-methoxy-9'-(5-methoxypyridin-3-yl)-2'-oxo- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (70 mg, 0.157 mmol) in THF/EtOH/H20 (2 mL /1 mL /0.5 mL) was added LiOH (39.5 mg, 0.94 mmol). After stirred at r.t for 1 hr, the resulting mixture was neutralized with 1 N HCI, then basified with NH3/H2O, extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (23 mg, 35% yield ) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 11 .94 (br, 1 H), 8.86 (s, 1 H), 8.36 (s, 1 H), 8.30 (d, J =2.5 Hz,1 H), 7.69 (s, 1 H),7.66 (s, 1 H), 7.55 (s, 1 H), 7.52 (s, 1 H), 3.94 (s, 3H), 3.89 (s, 3H), 3.36 (s, 2H), 2.72 - 2.62 (m, 2H), 2.10 - 2.02 (m, 2H), 1.95 - 1 .86 (m, 2H). LCMS (ESI) m/z: 419.3 (M + 1)+.
[001156] Example 99
[001157] 10'-Methoxy-2'-oxo-9'-(pyridin-3-yl)-2',7'-dihvdrospiro[cyclobutane-
Figure imgf000239_0002
[001158] Step 1 : Ethyl 10l-methoxy-2'-oxo-9l-(pyridin-3-yl)-2l.7l- dihvdrospir[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000239_0003
[001159] A mixture of ethyll 0'-methoxy-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate(100mg,0.205mmol), pyridin-3-ylboronic acid (45.5 mg, 0.369 mmol), Pd(dppf)CI2 (17 mg, 0.021 mmol), CH3COOK (60 mg, 0.62 mmol) in 1 ,4-dioxane(1 mL) and H20(1 drop) was stirred at 80°C under N2 atmosphere overnight. The mixture was concentrated, the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to afford the title compound (40 mg, 47% yield) as a yellow solid. LCMS (ESI) m/z: 417.3 (M + 1)+.
[001160] Step 2: l O'-Methoxy^'-oxo-g'-foyridin-S-vn^'J1- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000240_0001
[001161] To a solution of ethyl 10'-methoxy-2'-oxo-9'-(pyridin-3-yl)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (40 mg, 0.096 mmol) in THF/EtOH/H20(2 mL /1 mL /0.5 mL) was added LiOH (24.2 mg, 0.576 mmol). After stirred at r.t for 1 hr, the resulting mixture was neutralized with 1 N HCI, then basified with NH3/H2O , extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (12 mg, 32.2% yield ) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 1 1 .94 (br, 1 H), 8.86 (s, 1 H), 8.76 (s, 1 H), 8.58 (s, 1 H), 7.99 (d, J = 6.3 Hz,1 H), 7.68 (d, J =13.0 Hz, 2 H), 7.51 (s, 2H), 3.94 (s, 3H), 3.36 (s, 2H), 2.72 - 2.65 (m, 2H), 2.1 1 - 2.01 (m, 2H), 1 .96 - 1 .86(m, 2H). LCMS (ESI) m/z: 389.3 (M + 1)+.
[001162] Example 100
[001163] 10l-Methoxy-2'-oxo-9l-(1 Η-ρνΓ3ζοΙ-4-νΙ)-2'.7'- dihvdrospiro[cvclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000240_0002
[001164]
pyrazole-1 -carboxylate
Figure imgf000240_0003
[001165] To a mixture of 4-pyrazoleboronic acid pinacol ester (0.485 g, 2.5 mmol) and DMAP (0.153 g, 1.25 mmol) in MeCN (12.5 mL) was added di-te/ -butyl dicarbonate (0.709 g, 3.25 mmol). The resulting mixture was stirred at rt for 18 h before it was concentrated under reduced pressure. The obtained residue was purified by flash column chromatography (silica gel, 0-10% EtOAc in petroleum ether) to afford the title compound (0.473 g, 64% yield) as a white solid. Ή NMR (400 MHz, CDCI3) δ 8.38 (s, 1 H), 7.92 (s, 1 H), 1 .64 (s, 9H), 1.33 (s, 12H).
[001166] Step 2: Ethyl 10'-methoxy-2'-oxo-9'-(((trifluoromethyl')sulfonyl')oxyV 2',7'-dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylate
Figure imgf000241_0001
[001167] To a solution of ethyl 10'-methoxy-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate (122 mg, 0.25 mmol), fe/ -butyl 4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)-1 H-pyrazole-1 -carboxylate (147 mg, 0.5 mmol), Pd(OAc)2 (5.6 mg, 0.025 mmol), PPh3 (26.2 mg, 0.1 mmol) and KF (43.6 mg, 0.75 mmol) was added 1 ,4-dioxane/H20 (1.25 mL/0.31 mL). The resulting mixture was stirred at 100 °C for 32 h, cooled down to rt and concentrated under reduced pressure. The obtained residue was purified by preparative thin layer chromatography (silica gel, 0-6% MeOH in CH2CI2) to afford ethyl 9'-(1-(tert- butoxycarbonyl)-1 H-pyrazol-4-yl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylate (38 mg, 30% yield) as brown viscous oil. LCMS (ESI) m/z: 406.32 (M-Boc)+.
Figure imgf000241_0002
dihydrospirofcyclobutan -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000241_0003
[001169] To a solution of ethyl 9'-(1-(tert-butoxycarbonyl)-1 H-pyrazol-4-yl)-10'- methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (38 mg, 0.075 mmol) in CH2CI2 (0.75 mL) was added TFA (34 mg, 0.3 mmol). The resulting mixture was stirred at rt for 1 h before it was concentrated under reduced pressure. The obtained residue was dissolved in MeOH/H20 (0.5 mL/0.5 mL), followed by addition of LiOH (7.2 mg, 0.3 mmol). The resulting mixture was stirred at rt for 2 h, acidified with 1 N HCI to pH around 6 and extracted with CH2CI2 (3x2 mL). The combined organic layers were washed with brine, dried over Na2S04 and concentrated to give the crude product which was purified with reverse phase HPLC (C18, 0-90% MeCN in H2O with 0.1 % formic acid) to afford the title compound (7.4 mg, 26% yield) as a yellow powder. 1H NMR (400 MHz, DMSO-de) δ 13.08 (s, 1 H), 8.84 (s, 1 H), 8.42 (s, 1 H), 8.17 (br s, 2H), 7.76 (s, 1 H), 7.58 (s, 1 H), 7.56 (s, 1 H), 4.01 (s, 3H), 3.31 (s, 2H), 2.71 - 2.60 (m, 2H), 2.10 - 2.01 (m, 2H), 1 .96 - 1 .85 (m, 2H). LCMS (ESI) m/z: 378.3 (M + 1 )+.
[001170] Example 101
[001171] 9'-(Cvanomethyl)-10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000242_0001
[001172] Step 1 : 9'-(Cvanomethyl)-10'-methoxy-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000242_0002
[001173] To a mixture of ethyl 10'-methoxy-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate (146 mg, 0.3 mmol), B-4-isoxazolylboronic acid (50.8 mg, 0.45 mmol), Pd(OAc)2 (6.7 mg, 0.03 mmol), PPh3 (31 .5 mg, 0.12 mmol) and KF (52.3 mg, 0.9 mmol) was added 1 ,4-dioxane/H20 (1 .5 mL/0.38 mL). The resulting mixture was stirred at 100 °C for 22 h, cooled down to rt and diluted with H20 (1 .0 mL). The obtained mixture was filtered. To the collected filtrate was added LiOH (21 .6 mg, 0.9 mmol), and the resulting mixture was stirred at rt for 2 h before it was acidified with 1 N HCI to pH around 2. The obtained mixture was extracted with CH2CI2 (2x3 mL), and the combined organic layers were washed with brine, dried over Na2S04, filtered and concentrated under reduced pressure. The obtained residue was purified by reverse phase HPLC (C18, 0-90% MeCN in H20 with 0.1 % formic acid) to afford the title compound (7.5 mg, 7% yield over two steps) as a white powder. 1H NMR (400 MHz, DMSO-de) δ 8.84 (s, 1 H), 7.61 (s, 2H), 7.44 (s, 1 H), 3.98 (s, 3H), 3.96 (s, 2H), 3.33 (s, 2H), 2.70 - 2.60 (m, 2H), 2.04 - 1 .98 (m, 2H), 1 .97 - 1 .83 (m, 2H). LCMS (ESI) m/z: 351 .2 (M + 1)+.
[001174] Example 102
[001175] 9l-(4-Carboxybutyl)-10l-methoxy-2'-oxo-2l.7l-
Figure imgf000243_0001
[001177] To a suspension of (2-carboxyethyl)triphenylphosphonium bromide (292 mg, 0.681 mmol) in THF (5 mL), was added NaHMDS (2M, 0.85 mL,1 .7mmol) dropwise at 0°C. The mixture was stirred at 0°C for 45 min, ethyl 9'-formyl-10'-methoxy -2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (250 mg, 0.681 mmol) in THF (5 mL) was added dropwise at 0°C, and the mixture was stirred at room temperature overnight. The resulting mixture was acidified with 1 N HCI at 0°C, ethyl acatate and water were added, the organic layer was separated, dried over Na2S04, concentrated and the residue was purified by flash chromatography (silica gel, 0~20% MeOH in DCM) to give the title compound (170 mg, 47% yield over two steps) as a yellow solid. LCMS (ESI) m/z: 438.2 (M + 1)+ .
[001178] Ste p 2 : 5-(3'-(Ethoxycarbonyl)-10'-methoxy-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinl-9'-yl)pentanoic acid
Figure imgf000243_0002
[001179] To a solution of (E)-5-(3'-(ethoxycarbonyl)-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinolin]-9'-yl)pent-4-enoic acid(120 mg, 0.27 mmol) in EtOH (8 ml_) was added palladium ( 50 mg, 5% on carbon (wetted with ca. 55% Water)), the resulting mixture was stirred at r.t under H2 overnight. Palladium was removed by filtering, the filtrate was concentrated to afford the title compound (120 mg, 99.6% yield). LCMS (ESI) m/z: 440.3 (M + 1)+.
[001180] Step 3: 9'-(4-Carboxybutyl)-10'-methoxy-2'-oxo-2'.7'- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000244_0001
[001181] To a solution of 5-(3'-(ethoxycarbonyl)-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinolin]-9'-yl)pentanoic acid (120 mg, 0.273 mmol) in THF/EtOH/H20(2 mL /1 mL /0.5 mL) was added LiOH (68.8 mg, 1 .638 mmol). After stirred at r.t for 1 hr, the resulting mixture was neutralized with 1 N HCI, extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0- 100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (27 mg, 24.1 % yield ) as a white powder. Ή NMR (400 MHz, DMSO-d6) 512.00 (br, 1 H),1 1 .88 (br, 1 H), 8.82 (s, 1 H), 7.53 (s, 1 H), 7.48 (s, 1 H), 7.23 (s,1 H), 3.91 (s, 3H), 3.27 (s, 2H), 2.67 - 2.59 (m, 4H), 2.24 (t, J = 6.9 Hz, 2H), 2.03 - 1 .97 (m, 2H), 1 .93 - 1 .85 (m, 2H), 1 .58 - 1 .51 (m, 4H). LCMS (ESI) m/z: 412.3 (M + 1)+.
[001182] Example 103
[001183] 9'-(5-Amino-5-oxopentyl)-10'-methoxy-2'-oxo-2'.7'- dihvdrospiro[cvclobutane- '-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000244_0002
[001184] Step 1 : Ethyl 9'-(5-amino-5-oxopentviyi 0'-methoxy-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000245_0001
[001185] A mixture of 5-(3'-(ethoxycarbonyl)-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinolin]-9'-yl)pentanoic acid(45 mg, 0.1 mmol), NH4CI (27.4 mg, 0.5 mmol), HBTU (57 mg, 0.15 mmol) and DIPEA (124 uL, 0.7 mmol) in DMF (2 mL) was stirred at r.t overnight. The mixture was concentrated and the residue was purified by flash chromatography (silica gel, 0~20% MeOH in DCM) to give the title compound (45 mg, 100% yield) as a yellow solid. LCMS (ESI) m/z: 439.4 (M + 1 )+.
[001186] Step 2: 9'-(5-Amino-5-oxopentyl)-10'-methoxy-2'-oxo-2'.7'- dihydrospirofcyclobutan '-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000245_0002
[001187] To a solution of ethyl 9'-(5-amino-5-oxopentyl)-10'-methoxy-2'-oxo- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate(45 mg, 0.102 mmol) in THF/EtOH/H20(2 mL /1 mL /0.5 mL) was added LiOH (26 mg, 0.616 mmol). After stirred at r.t for 1 hr, the resulting mixture was neutralized with 1 N HCI, extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0- 100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (13.2 mg, 31 .4% yield ) as a white powder. Ή NMR (400 MHz, DMSO-d6) 512.00 (br, 1 H), 8.82 (s, 1 H), 7.53 (s, 1 H), 7.47 (s, 1 H), 7.22 (s, 2H), 6.68 (s, 1 H), 3.91 (s, 3H), 3.27 (s, 2H), 2.67 - 2.56 (m, 4H), 2.08 - 1 .89 (m, 6H), 1 .58 - 1 .49 (m, 4H). LCMS (ESI) m/z: 41 1 .3 (M + 1)+.
[001188] Example 104
[001189] 9'-(4-Cvanobutyl)-10'-methoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane-
Figure imgf000245_0003
[001190] Step 1 : Ethyl 9'-(4-cvanobutyl)-10'-methoxy-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000246_0001
[001191] To a solution of ethyl 9'-(5-amino-5-oxopentyl)-10'-methoxy-2'-oxo- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (1 10 mg, 0.25 mmol),and pyridine (40 mg, 0.5 mmol) in dioxane (5 ml_) and DCM (5 ml_) at 0°C, was added TFAA(59 mg, 0.28 mmol) dropwise, the mixture was stirred at r.t for 2 h. Then the mixture was poured into H20, extracted with DCM, the organic layer was separated, dried over Na2S04, concentrated and the residue was purified by flash chromatography (silica gel, 0-20% MeOH in DCM) to give the title compound (100 mg, 95% yield) as a yellow solid. LCMS (ESI) m/z: 421 .4 (M + 1)+.
[001192] Step 2: 9l-(4-Cvanobutyl)-10l-methoxy-2'-oxo-2l.7l- dihvdrospiro[cvclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000246_0002
[001193] To a solution of ethyl 9'-(4-cyanobutyl)-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.238 mmol) in THF/EtOH/H20(2 mL /1 mL /0.5 mL) was added LiOH (60 mg, 1 .43 mmol). After stirred at r.t for 1 hr, the resulting mixture was neutralized with 1 N HCI, extracted with DCM/i-PrOH. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by reverse phase HPLC (C18, 0- 100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (34.5 mg, 37% yield ) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 12.20 (br, 1 H), 8.82 (s, 1 H), 7.54 (s, 1 H), 7.49 (s, 1 H), 7.25 (s, 1 H), 3.92 (s, 3H), 3.27 (s, 2H), 2.69 - 2.59 (m, 4H), 2.53 (t, J= 6.9 Hz, 2H), 2.05 - 1 .84 (m, 4H), 1 .70 - 1 .56 (m, 4H). LCMS (ESI) m/z: 393.1 (M + 1 )+.
[001194] Example 105
[001195] 2'-Oxo-2'.7'.10'.1 1 '-tetrahvdrospiro[cvclobutane-1 .6'-[1 .4ldioxino[2.3- qlpyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000247_0001
[001196] Step 1 : Ethyl 2'-oxo-2'.7'.10'.1 1 '-tetrahvdrospiro[cvclobutane-1 .6'- [1 ,4ldioxino[2,3-qlpyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000247_0002
[001197] To a 50 ml. round flask, ethyl 9',10'-dihydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.27mmol), 3 equivalents of K2C03 (1 15 mg, 0.51 mmol) were dissolved in 10 mL acetonitrile at room temperature overnight. When the reaction was finished, the mixture was washed with water and extracted by EtOAc (10 mL * 3). The organic layers were combined and washed with brine, dried over anhydrous Na2S04, and concentrated by vacuum. The residue was purified by TLC plate and gave the title compound ethyl 2'-oxo-2',7',10',1 1 '- tetrahydrospiro[cyclobutane-1 ,6'-[1 ,4]dioxino[2,3-g]pyrido[2,1 -a]isoquinoline]-3'-carboxylate 30 mg (0.081 mmol , 30 % yield) as a white solid and used directly in the next step. LCMS (ESI) m/z: 368.3 (M + 1)+.
[001198] Step 2: 2'-Oxo-2'.7'.10'.1 1 '-tetrahvdrospiro[cvclobutane-1 .6'- [1 ,4ldioxino[2,3-glpyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000247_0003
[001199] To a 50 mL flask equipped with a stirring bar, 30 mg (0.081 mml) of ethyl 2'-oxo-2',7',10',1 1 '-tetrahydrospiro[cyclobutane-1 ,6'-[1 ,4]dioxino[2,3-g]pyrido[2,1 - a]isoquinoline]-3'-carboxylate and 3 equivalents of lithium hydroxide (6 mg, 0.24 mmol) were dissolved in THF/H20 3:1 (4 mL) at room temperature. When the reaction was finished, the resulting mixture was acidified by HCI and stirred for 30 minutes. After that, the mixture was added water and extracted with EtOAc (10 mL*3). The organic layers were combined and washed with brine, dried over anhydrous Na2S04, and concentrated by vacuum. The residue was purified by TLC plate and 5 mg (0.015 mmol, 18 % yield) 2'-oxo- 2',7',10',1 1 '-tetrahydrospiro[cyclobutane-1 ,6'-[1 ,4]dioxino[2,3-g]pyrido[2,1 -a]isoquinoline]-3'- carboxylic acid was obtained as a white solid. LCMS (ESI) m/z: 340.2 (M + 1)+. 1H NMR (400 MHz, DMSO-de) δ 8.78 (s, 1 H), 7.58 (s, 1 H), 7.26 (s, 1 H), 6.97 (s, 1 H), 4.44 - 4.31 (m, 2H), 4.31 - 4.21 (m, 2H), 3.24 (s, 2H), 2.62 (dd, J = 20.7, 9.7 Hz, 2H), 2.06 - 1 .95 (m, 2H), 1 .93 - 1 .80 (m, 2H).
[001200] Example 106
[001201] 3,3-Difluoro-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000248_0001
[001203] n-Butyllithium (2.5 M in hexanes) (3.65 mL, 9.12 mmol) was added dropwise to a -78 °C solution of diisopropylamine (1 .300 mL, 9.12 mmol) in tetrahydrofuran (THF) (20 mL). The mixture was allowed to warm to room temperature and stirred for 15 minutes. The mixture was cooled to -78 °C before methyl 3,3-difluorocyclobutane-1 - carboxylate (1325 mg, 8.83 mmol) was added dropwise. The mixture was warmed to 0 °C and stirred for 15 minutes before a solution of 4-(bromomethyl)-1 -methoxy-2-(3- methoxypropoxy)benzene (851 mg, 2.94 mmol) in tetrahydrofuran (THF) (7 mL) was added dropwise. The mixture was stirred an additional 15 minutes at 0 °C and then allowed to warm to room temperature and stirred for 15 minutes. The mixture was quenched with saturated ammonium chloride and extracted 2 times with ethyl acetate. The combined organic layers were washed with brine, and concentrated to give crude methyl 3,3-difluoro- 1 -(4-methoxy-3-(3-methoxypropoxy)benzyl)cyclobutanecarboxylate. LCMS (ESI) m/z: 359.3 (M+1)+.
Figure imgf000248_0002
[001205] A solution of lithium hydroxide monohydrate (1235 mg, 29.4 mmol) in water (5 mL) was added to a solution of methyl 3,3-difluoro-1 -(4-methoxy-3-(3- methoxypropoxy)benzyl)cyclobutane-1 -carboxylate (1055 mg, 2.94 mmol) in methanol (5 ml_) and tetrahydrofuran (THF) (5 ml_). The mixture was stirred at room temperature overnight and then concentrated to ~10 ml_. The mixture was acidified with 1 M hydrochloric acid and extracted 3 times with dichloromethane. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated. The residue was purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient). Fractions were concentrated and the residue lyophilized (acetonitrile / water) to give 3,3-difluoro-1 -(4-methoxy-3-(3- methoxypropoxy)benzyl)cyclobutane-1 -carboxylic acid (761 mg, 2.03 mmol, 69 % yield) as a light tan solid. LCMS (ESI) m/z: 345.3 (M+1)+.
Figure imgf000249_0001
[001207] Diphenyl phosphorazidate (0.572 ml_, 2.65 mmol) was added dropwise to a 0 °C stirring mixture of 3,3-difluoro-1 -(4-methoxy-3-(3- methoxypropoxy)benzyl)cyclobutane-1 -carboxylic acid (761 mg, 2.210 mmol) and triethylamine (0.370 ml_, 2.65 mmol) in toluene (12 ml_). The mixture was allowed to warm to room temperature and stirred for 15 minutes before being heated at 80 °C for 1 hour. The mixture was allowed to cool to room temperature before a mixture 5N hydrogen chloride (15 ml_, 75 mmol) and 1 ,4-dioxane (30 ml_) was added. The mixture was heated at 80 °C with vigorous stirring for 2.5 hours. The mixture was allowed to cool to room temperature and was extracted with ethyl acetate. The ethyl acetate layer was back- extracted 1 time with 1 N hydrochloric acid. The combined acidic aqueous layers were basified with 1 M sodium hydroxide and extracted 3 times with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give 3,3-difluoro-1 -(4-methoxy-3-(3-methoxypropoxy)benzyl)cyclobutan-1 -amine (510 mg, 1 .617 mmol, 73.2 % yield) as an oil. LCMS (ESI) m/z: 316.3 (M+1)+.
[001208] Step 4: N-(3.3-Difluoro-1 -(4-methoxy-3-(3- methoxypropoxy)benzvhcvclobutyl)formamide
Figure imgf000249_0002
[001209] Acetic formic anhydride (6.79M in formic acid) (1 .19 ml_, 8.09 mmol) was added dropwise to a 0 °C solution of 3,3-difluoro-1 -(4-methoxy-3-(3- methoxypropoxy)benzyl)cyclobutan-1 -amine (510 mg, 1 .62 mmol) and triethylamine (0.451 mL, 3.23 mmol) in dichloromethane (DCM) (15 mL). The mixture was stirred at 0 °C for 2 hours. The mixture was diluted with dichloromethane, washed with saturated sodium bicarbonate, washed with brine, dried over sodium sulfate, and concentrated to give crude
N-(3,3-difluoro-1 -(4-methoxy-3-(3-methoxypropoxy)benzyl)cyclobutyl)formamide (555 mg).
LCMS (ESI) m/z: 344.3 (M+1)+.
[001210]
spiro[cvclobutane-
Figure imgf000250_0001
[001211] N-(3,3-Difluoro-1 -(4-methoxy-3-(3- methoxypropoxy)benzyl)cyclobutyl)formamide (555 mg, 1 .62 mmol) in acetonitrile (15 mL) was cooled in an ice bath before phosphorus oxychloride (0.181 mL, 1 .94 mmol) was added dropwise with stirring. The mixture was heated at 60 °C for 4 hours and concentrated. The residue was dissolved in ethyl acetate and cooled in an ice bath before water and 30%
NH4OH were added to bring the pH =~1 1 . The organic layer was collected and the aqueous layer extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give 3,3-difluoro-7'-methoxy-6'-(3- methoxypropoxy)-4'H-spiro[cyclobutane-1 ,3'-isoquinoline] (503 mg, 1 .55 mmol, 96 % yield).
LCMS (ESI) m/z: 326.2 (M+1)+.
[001212] Step 6: Ethyl 3,3-difluoro-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo- 2',7'-dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000250_0002
[001213] 3,3-Difluoro-7'-methoxy-6'-(3-methoxypropoxy)-4'H- spiro[cyclobutane-1 ,3'-isoquinoline] (503 mg, 1 .546 mmol) and ethyl 2-(ethoxymethylene)-3- oxobutanoate (1 .345 mL, 7.73 mmol) in ethanol (6 mL) were heated at reflux overnight. Additional ethyl 2-(ethoxymethylene)-3-oxobutanoate (1 .35 mL, 7.73 mmol) was added and the mixture continued to heat at 80 °C for 4 hours. The mixture was concentrated and the residue dissolved in 1 ,2-eimethoxyethane (DME) (6 mL). p-Chloranil (380 mg, 1 .55 mmol) was added and the mixture heated at 85 C for 2 hours. The mixture was cooled in an ice bath, concentrated, and the residue purified by silica chromatography eluting with a gradient of 0% to 100% ethyl acetate in dichloromethane. Fractions were concentrated to give ethyl 3,3-difluoro-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylate (280 mg, 0.574 mmol, 37 % yield) as a tan solid.
LCMS (ESI) m/z: 464.3 (M+1)+.
[001214] Step 7: 3,3-Difluoro-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000251_0001
[001215] A solution of lithium hydroxide monohydrate (254 mg, 6.04 mmol) in water (1 .67 mL) was added to a solution of ethyl 3,3-difluoro-10'-methoxy-9'-(3- methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (280 mg, 0.604 mmol) in methanol (5 mL) and the mixture heated at 60 °C for 2 hours. The mixture was allowed to cool to room temperature, diluted with water, acidified with 1 M hydrochloric acid, and extracted 3 times with dichloromethane. The combined organic layers were washed with brine and concentrated. The residue was purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient). Fractions were concentrated. The residue was lyophililzed (acetonitrile / water) to give 3,3-difluoro-10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (53 mg, 0.1 14 mmol, 18.9 % yield) as a tan solid. Ή NMR (400 MHz, DMSO-c/6) δ ppm 1 .91 - 2.02 (m, 2 H) 2.77 - 2.94 (m, 2 H) 3.29 (s, 5 H) 3.36 - 3.51 (m, 4 H) 3.85 (s, 3 H) 4.08 (t, J=6.44 Hz, 2 H) 7.06 (s, 1 H) 7.44 (s, 1 H) 7.47 (s, 1 H) 8.73 (s, 1 H). 22 of 23 protons found. LCMS (ESI) m/z: 436.3 (M+1)+.
[001216] Example 107
[001217] 3-(6-(Diisobutylamino)-5-((3-(trifluoromethyl)-1 .2.4-thiadiazol-5-yl) amino)pyridin-3-yl)butanoic acid
Figure imgf000251_0002
[001218] Step 1 : 2.2-Difluoropropane-1 ,3-diol
HC DH
F F
[001219] To a solution of diethyl difluoropropanedioate (2.0 g, 10mmol) in THF (100 mL) at 0 °C is added L1AIH4 (4 equiv., 1 .52 g). The reaction mixture is warmed to room temperature and stirred for 16 h. Water (100 mL) is added carefully and the pH is adjusted to 3 by adding 1 M HCI solution. The water is removed in vacuo and the residue is extracted with EtOAc (3x100 mL). The organic layers are combined, dried (Na2S04) and concentrated to afford the title compound (1 .7 g, crude).
[001220] Step 2: 2,2-Difluoro-3-hvdroxypropyl 4-methylbenzenesulfonate
HO^x^OTs
F F
[001221] To a solution of 2,2-difluoropropane-1 ,3-diol (1 .22 g, 1 1 mmol) in DCM was added triethylamine(3.3 g, 33mmol ) and 4-toluene sulfonyl chloride (1 .82 g, 8mmol). The resulting mixture was stirred for 12 h at room temperature. To the mixture was added water and then the organic layer was separated, washed with brine, dried over anhydrous Na2S04 and concentrated to give 235 mg crude of 2,2-difluoro-3-hydroxypropyl 4-methylbenzenesulfonate as a yellow oil. LCMS (ESI) m/z: 267.2 (M + 1 )+.
[001222] Step 3: Ethyl 9'-(2,2-difluoro-3-hvdroxypropoxy)-10'-methoxy-2'-oxo- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000252_0001
[001223] To a solution of ethyl 9'-hydroxy-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (123 mg, 0.35 mmol) in DMF(2 mL) was added potassium carbonate (98 mg, 0.7 mmol) and 2, 2-difluoro-3- hydroxypropyl 4-methylbenzenesulfonate (92 mg, 0.35 mmol). Then the resulting mixture was heated at 90 °C for 3 h. After then, cooled to room temperature, the dark-brown mixture was poured into water (10 mL) and extracted with DCM (5 mL*3). The organic layers were combined and washed with brine, dried over anhydrous Na2S04, and concentrated by vacuum to give 20 mg ethyl 9'-(2,2-difluoro-3-hydroxypropoxy)-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate, which was used directly for the next step. LCMS (ESI) m/z: 450.3 (M + 1 )+.
[001224] Step 4: 9'-(2,2-Difluoro-3-hvdroxypropoxy)-10'-methoxy-2'-oxo-2',7'- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000252_0002
[001225] To a solution of ethyl 9'-(2,2-difluoro-3-hydroxypropoxy)-10'- methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (20 mg, 0.06 mmol) in THF(4 mL) and ethanol (1 mL) was added 2.0 M LiOH (1 mL) aqueous solution at room temperature. The resulting mixture was stirred for 1 h, and then mixture was extracted with DCM (10 mLx 3). The combined organic layers were washed with water and brine, dried over anhydrous sodium sulfate, and concentrated by vacuum to give crude product 9'-(2,2-difluoro-3-hydroxypropoxy)-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid, which was purified by reverse phase HPLC to give the final compound 19 mg. LCMS (ESI) m/z: 422.2 (M + 1)+. 1 H NMR (400 MHz, DMSO-d6) δ 8.80 (s, 1 H), 7.55 (s, 1 H), 7.46 (s, 1 H), 7.18 (s, 1 H), 5.68 (t, J = 6.0 Hz, 1 H), 4.41 (t, J = 12.7 Hz, 2H), 3.90 (s, 3H), 3.78 (td, J = 13.7, 5.5 Hz, 2H), 3.28 (s, 2H), 2.64 (dd, J = 19.8, 7.9 Hz, 2H), 2.06 - 1.98 (m, 2H), 1.89 (dd, J = 12.4, 7.7 Hz, 2H).
[001226] Example 108
[001227] 10l-Methoxy-9l-(4-methoxybutyl)-2'-oxo-2l.7l- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000253_0001
[001228] Stepl : Ethyl 9'-(4-hvdroxybut-1 -vn-1 -yl)-10'-methoxy-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000253_0002
[001229] To a mixture of ethyl 10'-methoxy-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]- 3'-carboxylate (200 mg, 0.410 mmol) , Pd(PPh3)2CI2 (143.8 mg,0.205mmol),K2CO3 (113.2 mg,0.82mmol) , Cul (7.8 mg, 0.041 mmol) in DMF (10 ml), but-3-yn-1-ol (172.4 mg, 2.46 mmol) was added. The resulting mixture was stirred at 60 °C for 3 h. Then the mixture was diluted with EtOAc (20mL) and H20 (30 mL), and then extracted with EtOAc (50mLx2). The combined organic solution was washed with brine, dried over anhydrous Na2S04 and concentrated. The residue was purified by column chromatography (silica gel,0~10%MeOH in DCM) to afford ethyl 9'-(4-hydroxybut-1 -yn-1-yl)-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (150 mg, 89.8% yield) as a yellow solid. LCMS (ESI) m/z: 408.3 (M + 1)+.
[001230] Step 2: Ethyl 9'-(4-hvdroxybutyl)-10'-methoxy^'-oxo^'J1- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000254_0001
[001231] A mixture of ethyl 9'-(4-hydroxybut-1 -yn-1-yl)-10'-methoxy-2'-oxo- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (150 mg, 0.368 mmol) and10% wet Pd/C (100 mg) in MeOH (10 ml_) was stirred under hydrogen atmosphere at r. t. for 1 h.The mixture was filtered and the filtrate was concentrated under reduced pressure to afford ethyl 9'-(4-hydroxybutyl)-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (120 mg, 79.2% yield) as a yellow oil. LCMS (ESI) m/z: 412.3 (M + 1)+.
[001232] Step 3: 10'-Methoxy-9'-(4-methoxybutyl)-2'-oxo-2'.7'- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000254_0002
[001233] To a mixture of NaH (21 mg, 0.875mmol) in THF(5 ml), ethyl 9'-(4- hydroxybutyl)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1- a]isoquinoline]-3'-carboxylate (120 mg, 0.292 mmol) was added. After stirring for 5 minutes, Mel (248.4 mg, 1 .75mmol) was added. The resulting mixture was stirred at 0 °C for 2 h. The reaction mixture was quenched by adding cold NhUCI solution and acidified to ph~5 with 1 N HCI. Then the resulting mixture was diluted with EtOAc (20ml_) and H20 (30 ml_), and then extracted with EtOAc (50ml_x2). The combined organic solution was dried over anhydrous Na2S04 and concentrated. The residue was purified by Gilson (C18
column,20%~100%MeCN in H20, with 0.1 % formic acid in H20) to give 10'-methoxy-9'-(4- methoxybutyl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'- carboxylic acid (27.5 mg, 20.2% yield) as a light pink solid. LCMS (ESI) m/z: 398.3 (M + 1)+. Ή NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1 H), 7.53 (s, 1 H), 7.48 (s, 1 H), 7.23 (s, 1 H), 3.91 (s, 3H), 3.35 - 3.32 (m, 2H), 3.27 (s, 2H), 3.21 (s, 3H), 2.69 - 2.58 (m, 4H), 2.02 - 1.96 (m, 2H), 1.93 - 1.82 (m, 2H), 1.62 - 1 .49 (m, 4H). [001234] Example 109
[001235] 10-(DifluoromethylV9-(furan-3-ylV6-isopropyl-6-methyl-2-oxo-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000255_0001
[001236] Step 1 : Ethyl 9-(benzyloxy)-6-isopropyl-6-methyl-2-oxo-10-vinyl-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000255_0002
[001237] To a solution of ethyl 9-(benzyloxy)-10-bromo-6-isopropyl-6-methyl- 2-0X0-6, 7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (1 .0 g, 1 .96 mmol) in EtOH (15 mL) was added Potassium Vinyltrifluoroborate (525 mg, 3.92 mmol), Pd(Ph3)4 (226 mg, 0.2 mmol) and Na2C03 (623 mg, 5.88 mmol) under N2 atmosphere. The reaction mixture was stirred overnight at 85 °C. After the reaction, the reaction mixture was added to water 50 mL, and extracted with ethyl acetate. Dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure, an oily liquid was obtained. This was dissolved in a small amount of ethyl acetate, packed to silica gel chromatography, subjected to column purification using hexane/ethyl acetate as an eluent. The solvent of the obtained solution can be distilled off under reduced pressure to afford the title compound (800 mg, 89% yield), LCMS (ESI) m/z calcd for C29H31 NO4: 457.23. Found: 458.23 (M+1)+.
[001238] Step 2: Ethyl 9-(benzyloxy)-10-formyl-6-isopropyl-6-methyl-2-oxo- 6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000255_0003
[001239] Ethyl 9-(benzyloxy)-6-isopropyl-6-methyl-2-oxo-10-vinyl-6,7-dihydro- 2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (800 mg, 1 .75 mmol), Nal04 (1 .5 g, 7.0 mmol) and distilled water (3 mL) were sequentially added to a THF (12 mL) solution of the K2040s (66 mg, 0.18 mmol) under an inert atmosphere. The mixture was stirred at 25 °C for 0.5 h. The progress of the reaction was monitored by LCMS. After the reaction was finished, the reaction mixture was extracted by DCM and the solution was evaporated under vacuo. The residue was purified by column chromatography on silica gel (DCM: MeOH) to afford the title compound (680 mg, 85% yield). LCMS (ESI) m/z calcd for C28H29NO5: 459.20. Found: 460.21 (M+1)+.
[001240] Step 3: Ethyl 10-formyl-9-hvdroxy-6-isopropyl-6-methyl-2-oxo-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000256_0001
[001241] To a solution of ethyl 9-(benzyloxy)-10-formyl-6-isopropyl-6-methyl- 2-0X0-6, 7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (6 g, 13 mmol) in TFA. The mixture reaction was stirred at 65 °C. After the reaction was finished, the mixture was concentrated. The residue was distilled in DCM, washed with K2CO3, dried over MgS04 and concentrated. The crude was purified with flash chromatography to afford the title compound (3.6 g, 75% yield). LCMS (ESI) m/z calcd for C21 H23NO5: 369.16. Found: 370.16 (M+1)+.
[001242] Step 4: Ethyl 10-formyl-6-isopropyl-6-methyl-2-oxo-9-
Figure imgf000256_0002
[001243] To a solution of ethyl 10-formyl-9-hydroxy-6-isopropyl-6-methyl-2- oxo-6, 7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (3.3 g, 8.9 mmol) in DCM was added PhN(Tf)2 (3.8 g, 10.7 mmol), DMAP (54 mg, 0.45 mmol), TEA (2.7 g, 26.7 mmol). The reaction mixture was stirred at room temperature for 2 h. Then, the mixture reaction was filtered, and the residue was washed with EtOAc. The filtered was evaporated. The crude product was purified with flask chromatography (Column material: 20 g silica gel. Solvent: Ethyl acetate: iso-hexane = 1 :19) to afford the title compound (2.2 g, 49% yield). LCMS (ESI) m/z: 502.1 (M + 1)+. [001244] Step 5: Ethyl 10-formyl-9-(furan-3-yl)-6-isopropyl-6-methyl-2-oxo-6,7- dihvdro-2/-/-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000257_0001
[001245] Under a stream of nitrogen, ethyl 10-formyl-6-isopropyl-6-methyl-2- oxo-9-(((trifluoromethyl)sulfonyl)oxy)-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (220 mg, 0.44 mmol), furan-3-ylboronic acid (74 mg, 0.66 mmol), Pd(PPh3)4 (51 mg, 0.044 mmol), Cs2C03 (434 mg, 1 .32 mmol) and toluene (5 mL) were placed into a single round bottom flask, then stirred for 1 h at 100°C. After removed the solvent, the residue was purified by flash chromatography (silica gel, 0~30% MeOH in DCM) to afford the title compound. (160 mg, 87% yield) as a light yellow solid . LCMS (ESI) m/z: 420.4 (M + 1)+.
[001246] Step 6: Ethyl 10-(difluoromethyl)-9-(furan-3-yl)-6-isopropyl-6-methyl- 2-oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000257_0002
[001247] At 0°C, to a solution of ethyl 10-formyl-9-(furan-3-yl)-6-isopropyl-6- methyl-2-oxo-6,7-dihydro-2/-/-pyrido[2,1 -a]isoquinoline-3-carboxylate (160 mg, 0.38 mmol) in anhydrous DCM (3 mL) was added a solution of DAST (490 mg, 3.04 mmol) in DCM (1 mL) dropwise. After stirred at the same temperature for 4 hours, the reaction was quenched with sat.NaHC03, extracted with 15% IPA in DCM, the organic layers were dried over Na2S04, filtered and concentrated. The residue was purified by reverse phase HPLC (Gilson, C18, 20%~100% MeCN in water with 0.1 % formic acid) to afford the title compound. (65 mg, 39% yield) as a white solid. LCMS (ESI) m/z: 442.3 (M + 1)+.
[001248] Step 7: 10-(Difluoromethyl)-9-(furan-3-yl)-6-isopropyl-6-methyl-2- oxo-6, 7-dihydro-2/-/-pyrido[2, 1 -alisoquinoline-3-carboxylic acid
Figure imgf000258_0001
[001249] To a solution of ethyl 10-(difluoromethyl)-9-(furan-3-yl)-6-isopropyl-6- methyl-2-oxo-6,7-dihydro-2/-/-pyrido[2,1 -a]isoquinoline-3-carboxylate (10 mg, 0.023 mmol) in THF (1.5 mL) was added LiOH (1 .7 mg, 0.069 mmol) in water (0.5 mL) at room temperature After stirred at room temperature for 15 minutes. The reaction mixture was acidified to pH~6 with 1 N HCI and extracted with15% IPA in DCM twice. The combined organic layers were dried over Na2S04. Solvent were removed and the residue was purified by reverse phase HPLC (C18, 20-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (4 mg, 25% yield) as a white solid. 1H NMR (400 MHz, CDCI3) δ 15.78 (s, 1 H), 8.72 (s, 1 H), 8.17 (s, 1 H), 7.67 (s, 1 H), 7.59 (t, J = 1.6 Hz, 1 H), 7.34 (s, 1 H), 7.32 (s, 1 H), 6.85 - 6.58 (m, 2H), 3.24 (s, 2H), 1.94 - 1.89 (m, 1 H), 1.73 (s, 3H), 0.88 (d, J = 6.8 Hz, 3H), 0.75 (d, J = 6.8 Hz, 3H). LCMS (ESI) m/z: 414.1 (M + 1)+.
[001250] Example 110
[001251] 10-(Difluoromethyl)-6-isopropyl-9-methoxy-6-methyl-2-oxo-6,7- dihydro-2/-/-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000258_0002
[001252] Step 1 : Ethyl 10-formyl-6-isopropyl-9-methoxy-6-methyl-2-oxo-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000258_0003
[001253] To a mixture of ethyl 10-formyl-9-hydroxy-6-isopropyl-6-methyl-2- oxo-6, 7-dihydro-2H-pyrido [2, 1 -a] isoquinoline-3-carboxylate (360 mg, 0.98 mmol) in DMF (10 mL) were added Mel (415 mg, 2.93 mmol). The mixture was stirred at 50 °C for 1 h. The mixture was diluted by ice water (100 mL), filtered and the filter cake was washed by water (20 mL x 3) dried under vacuum to give the crude compound (370 mg, 99% yield) as a yellow solid. LCMS (ESI) m/z: 384.2 (M + 1)+ .
[001254] Step 2: Ethyl 10-(difluoromethyl)-6-isopropyl-9-methoxy-6-methyl-2- oxo-6, 7-dihvdro-2H-pyr -alisoquinoline-3-carboxylate
Figure imgf000259_0001
[001255] To a solution of ethyl 10-formyl-6-isopropyl-9-methoxy-6-methyl-2- oxo-6, 7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (350 mg, 0.913 mmol) in anhydrous DCM (5 mL) was added dropwise DAST (2.94 g, 18.26 mmol) in anhydrous DCM at 0°C under N2 atmosphere. The reaction mixture was stirred at 0°C for 1 h, the mixture was poured into sat.NaHC03 solution (20 mL) with ice. The solvent was extracted with DCM twice, the combined organic phases were dried over Na2S04, filtered and concentrated. The residue was purified by prep, reverse phase HPLC (Ci8, 10%-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (160 mg, 43.1 % yield). LCMS (ESI) m/z: 406.6 (M +
[001256] Step 3: 10-(Difluoromethyl)-6-isopropyl-9-methoxy-6-methyl-2-oxo- 6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000259_0002
[001257] To a solution of ethyl 10-(difluoromethyl)-6-isopropyl-9-methoxy-6- methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate ( 160 mg, 0.395 mmol) in CH3CN (4 mL) and H20 (1 mL) was added LiOH (47.28 mg, 1 .975 mmol). The reaction mixture was stirred at room temperature for 1 h. After acidified by 1 N HCI to PH 6, the reaction was extracted with DCM twice, the combined organic phases were dried over Na2S04, filtered and concentrated. The residue was purified by reverse phase HPLC (Gilson, C18 30%-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound as a white solid (97 mg, 65.1 % yield). LCMS (ESI) m/z: 378.2 (M + 1)+. Ή NMR (400 MHz, CDC ) δ 15.94 (s, 1 H), 8.67 (s, 1 H), 7.98 (s, 1 H), 7.17 (s, 1 H), 7.08 - 6.79 (m, 2H), 3.97(s, 3H), 3.20 (q, J= 16.2 Hz, 2H), 1 .92 (dt, J= 13.7, 6.8 Hz, 1 H), 1 .71 (s, 3H), 0.88 (d, J= 6.9 Hz, 3H) , 0.74 (d, J= 6.8 Hz, 3H). [001258] Example 111
[001259] 10-(Difluoromethyl)-9-ethvnyl-6-isopropyl-6-methyl-2-oxo-6,7-
Figure imgf000260_0001
[001261] To a solution of ethyl 10-formyl-6-isopropyl-6-methyl-2-oxo-9- (((trifluoromethyl)sulfonyl)oxy)-6,7-dihydro-2/-/-pyrido[2,1 -a]isoquinoline-3-carboxylate (360 mg, 0.72 mmol) in anhydrous DMF (5 mL) was added Pd(PPh3)2Cl2 (50.54 mg, 0.072 mmol), Cul (69.96 mg, 0.36 mmol) and K2CO3 (298.08 mg, 2.16 mmol) followed by ethynyltrimethylsilane (144.32 mg, 1 .44 mmol). The resulting mixture was heated at 50°C for 30 mins. After cooling down to room temperature, the mixture was portioned between DCM and sat. NH4CI, then was extracted with DCM twice. The combined organic phases were dried over Na2S04, filtered and concentrated to afford the crude title compound which was used to the next step directly. (310 mg, 95.9% yield). LCMS (ESI) m/z: 450.4 (M + 1)+.
[001262] Step 2: Ethyl 10-(difluoromethyl)-6-isopropyl-6-methyl-2-oxo-9- ((trimethylsilyl)ethvnyl)-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000260_0002
[001263] At 0°C, to a solution of ethyl 10-formyl-6-isopropyl-6-methyl-2-oxo-9- ((trimethylsilyl)ethynyl)-6,7-dihydro-2/-/-pyrido[2,1 -a]isoquinoline-3-carboxylate (310 mg, 0.689 mmol) in anhydrous DCM (4 mL) was added dropwise DAST (1 .777 g, 1 1 .024 mmol) in anhydrous DCM at 0°C under N2 atmosphere. The reaction mixture was stirred at 0°C for 30 mins, the mixture was poured into sat.NaHC03 (20 mL) at 0°C, then was extracted with DCM twice, the combined organic phases were dried over Na2S04, filtered and
concentrated to afford the crude title compound which was purified by flash chromatography (silica gel, 0-30% MeOH in DCM) to afford the title compound (140 mg, 43.1 % yield). LCMS (ESI) m/z: 472.3 (M + 1)+.
[001264] Step 3: 10-(Difluoromethyl)-9-ethvnyl-6-isopropyl-6-methyl-2-oxo- 6,7-dihvdro-2H-pyrido[2 -alisoquinoline-3-carboxylic acid
Figure imgf000261_0001
[001265] To a solution of ethyl 10-(difluoromethyl)-6-isopropyl-6-methyl-2-oxo- 9-((trimethylsilyl)ethynyl)-6,7-dihydro-2/-/-pyrido[2,1 -a]isoquinoline-3-carboxylate (40 mg, 0.085 mmol) in CH3OH (5 mL) was added K2C03 (58.65 mg, 0.425 mmol) at room temperature After stirring at room temperature for 1 h, the reaction mixture was
concentrated under reduced pressure at 45°C atmosphere, the residue was portioned between DCM and water, organic layer was separated and the aqueous layer was extracted with 15% IPA in DCM, the combined organic layers were dried over Na2S04, filtered and concentrate, the residue was purified by reverse phase HPLC (Cie 30% - 100% MeCN in H20 with 0.1 % formic acid) to afford the title compound as a white solid (3 mg, 9.5% yield). LCMS (ESI) m/z: 372.3 (M + 1)+.Ή NMR (400 MHz, CDCI3) δ 15.64 (s, 1 H), 8.71 (s, 1 H), 8.06 (s, 1 H), 7.49 (s, 1 H), 7.28 (s, 1 H), 7.03 (t, J=54.9 Hz, 1 H), 3.58 (s, 1 H), 3.20 (s, 2H), 1 .84 (dt, J =13.7, 6.9 Hz, 1 H), 1 .72 (s, 3H), 0.87 (d, J = 6.8 Hz, 3H) , 0.74 (d, J = 6.8 Hz, 3H).
[001266] Example 112
[001267] 10'-(Difluoromethyl)-9'-ethvnyl-2'-oxo-2',7'-dihvdrospiro[cyclobutane-
Figure imgf000261_0002
[001268] Step 1 : Ethyl 9'-(benzyloxyV2'-oxo-10'-vinyl-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000262_0001
[001269] To a solution of ethyl 9'-(benzyloxy)-10'-bromo-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (6.0 g, 12.1 mmol) in EtOH (120 mL) was added potassium vinyl trifluoroborate (3.2 g, 24.2 mmol), Pd(PPh3)4 (1 .4 g, 1 .21 mmol) and Na2C03 (3.85 g, 36.3 mmol) under N2 atmosphere. The reaction mixture was heated at 80 °C for overnight. After cooling to rt, the mixture was filtered, washed with DCM (100 mL). The filtrated was concentrated and the residue was purified by column chromatography using MeOH in DCM (1/15) to afford the title compound (4.6 g, 86% yield). LCMS (ESI) m/z 442.7 (M + 1 )+.
[001270] Step 2: Ethyl 9'-(benzyloxy)-10'-formyl^'-oxo^'J1- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000262_0002
[001271] To a solution of ethyl 9'-(benzyloxy)-2'-oxo-10'-vinyl-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (4.6 g, 10.4 mmol) in THF (100 mL) and H20 (25 mL) was added K20s04 2H20 (384 mg, 1 .04 mmol) at 0 °C, then Nal04 (8.9 g, 41 .6 mmol) was added in portions. The reaction mixture was stirred at rt for 2 h. After cooling to rt, the mixture was filtered, washed with DCM (100 mL). The filtrated was concentrated and the residue was purified by column chromatography using MeOH in DCM (1/15) to afford the title compound (4.6 g, 86% yield). LCMS (ESI) m/z: 444.3 (M + 1)+.
[001272] Step 3: Ethyl 10'-formyl-g'-hvdroxy^'-oxo^'J1- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000262_0003
[001273] A solution of ethyl 9'-(benzyloxy)-10'-formyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (3.9 g, 8.8 mmol) in TFA (80 mL) was heated at 70°C for 4 h. The solvent was removed under reduced pressure. The mixture was diluted with DCM (150 mL), washed with brine (80 mL x 3). The organic layer was dried over Na2S04, The solvent was removed under reduced pressure and the residue was triturated with EA/PE to afford the title compound (2.4 g, 77% yield). LCMS (ESI) m/z: 354.5 (M + 1)+.
[001274] Step 4: Ethyl 10'-formyl-2'-oxo-9'-(mrifluoromethvnsulfonyl)oxy)-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000263_0001
[001275] To a solution of ethyl 10'-formyl-2'-oxo-9'- (((trifluoromethyl)sulfonyl)oxy)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]- 3'-carboxylate (850 mg, 2.4 mmol) in DCM (20 mL) was added N,N-dimethylpyridin-4-amine (29 mg, 0.24 mmol), TEA (728 mg, 7.2 mmol) and 1 ,1 ,1 -trifluoro-N-phenyl-N- ((trifluoromethyl)sulfonyl)methanesulfonamide (1 .0 g, 2.9 mmol) under N2 atmosphere. The reaction mixture was stirred at rt for 4 h. The solvents were concentrated. The residue was purified by column chromatography using ethyl acatate to afford the title compound (650 mg, 73% yield). LCMS (ESI) m/z: 286.4 (M + 1)+.
[001276] Step 5: Ethyl 10'-formyl^'-oxo-g'^grimethylsilvDethvnvD^'J1- dihvdrospiro[cvclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000263_0002
[001277] To a solution of ethyl 10'-formyl-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (150 mg, 0.309 mmol) in anhydrous DMF (5 mL) was added Pd(PPh3)2CI2 (21 .76 mg, 0.031 mmol), Cul (30.12 mg, 0.155 mmol) and K2C03 (127.93 mg, 0.927 mmol), followed by
ethynyltrimethylsilane (61 .93 mg, 0.618 mmol). The resulting mixture was heated at 50°C for 30 mins. After cooling down to room temperature, the mixture was portioned between DCM and sat. NH4CI, then was extracted with DCM twice. The combined organic phases were dried over Na2S04, filtered and concentrated to afford the crude title compound which was used to the next step directly. (130 mg, 97.0% yield). LCMS (ESI) m/z: 434.3 (M + 1)+. [001278] Step 6: Ethyl 10'^difluoromethvi g'-ethynyl^'-oxo^'J1- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000264_0001
[001279] At 0°C, to a solution of ethyl 10'-formyl-2'-oxo-9'- ((trimethylsilyl)ethynyl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (130 mg, 0.30 mmol) in anhydrous DCM (2 mL) was added dropwise DAST (773.71 mg, 4.80 mmol). The resulting mixture was stirred at 0°C for 30 mins, the mixture was poured into sat.NaHC03 (20 mL) at 0°C, then was extracted with DCM twice, the combined organic phases were dried over Na2S04, filtered and concentrated to afford the crude title compound which was purified by reverse phase HPLC (Gilson, Ci8, 30%-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound (28 mg, 24.3% yield). LCMS (ESI) m/z: 384.3 (M + 1)+.
[001280] Step 7: I Q'^DifluoromethvD-g'-ethvnyl^'-oxo^'J'- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000264_0002
[001281] To a solution of ethyl 10'-(difluoromethyl)-9'-ethynyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate ( 28 mg, 0.07 mmol) in CH3CN (4 mL) and H20 (1 mL) was added LiOH (8.38 mg, 0.35 mmol). The reaction mixture was stirred at room temperature for 1 h. After acidified by 1 N HCI to PH 6, the reaction was extracted with DCM twice, the combined organic phases were dried over Na2S04, filtered and concentrated. The residue was purified by reverse phase HPLC (Gilson, C18, 30%-100% MeCN in H20 with 0.1 % formic acid) to afford the title compound as a white solid (7 mg, 26.9% yield). LCMS (ESI) m/z: 356.5 (M + 1 )+. 1H NMR (400 MHz, CDCI3) δ 15.64 (s, 1 H), 8.95 (s, 1 H), 8.03 (s, 1 H), 7.57 (s, 1 H), 7.23 (s, 1 H), 7.03 (t, J = 54.9 Hz, 1 H), 3.58 (s, 1 H), 3.32 (s, 2H), 2.71 - 2.62 (m, 2H), 2.21 - 2.12 (m, 2H), 2.10 - 1 .95 (m,2H).
[001282] Example 113
[001283] 10'-(Difluoromethyl)-9'-(neopentyloxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000265_0001
Figure imgf000265_0002
[001285] To a solution of ethyl 10'-formyl-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.28 mmol) in DMF (4 mL) was added neopentyl 4-methylbenzenesulfonate (127 mg, 0.84 mmol), K2C03 (1 16 mg, 0.84 mmol) and Nal (84 mg, 0.56 mmol). The reaction mixture was heated at 155 °C for 4 h at microwave irradiation. The mixture was poured into water (30 mL) and extracted with DCM (25 X 2 mL). The combined organic layers were washed with brine (30 X 4 mL). The solvents were concentrated. The residue was purified by Prep-TLC using MeOH in DCM (1/15) to afford the title compound (50 mg, 42% yield). LCMS (ESI) m/z: 424.5 (M + 1)+.
[001286] Step 2: Ethyl 10'-(difluoromethyl)-9'-(neopentyloxy)-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000265_0003
[001287] To a solution of ethyl 10'-formyl-9'-(neopentyloxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (50 mg, 0.12 mmol) in DCM (4 mL) was added dropwise DAST (77 mg, 0.48 mmol) at 0 °C under N2 atmosphere. The reaction mixture was stirred at rt for overnight. The mixture was diluted with DCM (30 mL), washed with sat.NaHC03 solution (20 mL) and brine (20 mL). The solvent was concentrated to afford the title compound (40 mg, 75% yield). LCMS (ESI) m/z: 446.4 (M + 1)+. [001288] Step 3: 10'-(Difluoromethyl)-9'-(neopentyloxy)-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000266_0001
[001289] To a solution of ethyl 10'-(difluoromethyl)-9'-(neopentyloxy)-2'-oxo-
2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate ( 40 mg, 0.09 mmol) in EtOH (4 mL) was added NaOH (14.4 mg, 0.36 mmol) in H20 (1 mL). The reaction mixture was stirred at rt for 2 h. The solvent was concentrated. The mixture was acidified to pH~1 by 1 N HCI and extracted with DCM (10 mL X 3). The combined organic layers were washed with brine (30 mL) and dried over Na2S04. The solvents were concentrated. The residue was purified by prep, reverse phase HPLC (C18, 20- 45 % acetonitrile in H20 with 0.1 % formic acid) to afford the title compound (11 mg, 29% yield) as a white powder. 1H NMR (400 MHz, DMSO-d6) δ 8.81 (s, 1 H), 8.13 (s, 1 H), 7.31 (d, J = 16.7 Hz, 2H), 7.09 (t, J = 54.7 Hz, 1 H), 3.84 (s, 2H), 3.42 (s, 2H), 2.71-2.63 (m,2H), 2.08 - 1.96 (m, 2H), 1.95 - 1.77 (m, 2H), 1.04 (s, 9H). LCMS (ESI) m/z: 418.5 (M + 1)+.
[001290] Example 114
[001291] 10l-(Difluoromethyl)-9'-(1 -methyl-1 H-pyrazol^-vD^'-oxo^'J'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000266_0002
[001292] Step 1 : Ethyl 10'-formyl-g'-d -methyl-1 H-pyrazol^-vD^'-oxo^'J'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000266_0003
[001293] To a solution of ethyl 10'-formyl-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (190 mg, 0.39 mmol) in 1 ,4-dioxane (8 mL) was added 1 -methyl-4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)- 1 H-pyrazole (98 mg, 0.47 mmol), Pd(PPh3)4 (45 mg, 0.04mmol) and Na2C03 (124 mg, 1 .17 mmol) under N2 atmosphere. The reaction mixture was heated at 90 °C for overnight. After cooling to rt, the mixture was filtered, washed with DCM (30 mL). The filtrated was concentrated and the residue was purified by pre-TLC using MeOH in DCM (1/15) to afford the title compound (94 mg, 58% yield). LCMS (ESI) m/z: 418.7 (M + 1)+.
[001294] Step 2: Ethyl 10'-(difluoromethyl)-9'-(1 -methyl-1 H-pyrazol-4-yl)-2'- oxo-2', 7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000267_0001
[001295] To a solution of ethyl 10'-formyl-9'-(1 -methyl- 1 H-pyrazol-4-yl)-2'-oxo- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (94 mg, 0.23 mmol) in DCM (4 mL) was added dropwise DAST (185 mg, 1 .15 mmol). at 0 °C under N2 atmosphere. The reaction mixture was stirred at rt for overnight. The mixture was diluted with DCM (30 mL), washed with sat.NaHC03 solution (20 mL) and brine (20 mL). The solvent was concentrated to afford the title compound (82 mg, 82% yield). LCMS (ESI) m/z: 440.6 (M + 1)+.
[001296] Step 3: 10'-(Difluoromethyl)-9'-(1 -methyl- 1 H-pyrazol-4-yl)-2'-oxo- 2',7'-dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000267_0002
[001297] To a solution of ethyl 10'-(difluoromethyl)-9'-(1 -methyl- 1 H-pyrazol-4- yl)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate ( 82 mg, 0.19 mmol) in EtOH (6 mL) was added NaOH (38 mg, 0.95 mmol) in H20 (1 .5 mL). The reaction mixture was stirred at rt for 2 h. The solvent was concentrated. The mixture was acidified to pH~1 by 1 N HCI and extracted with DCM (10 mL X 3). The combined organic layers were washed with brine (30 mL) and dried over Na2S04. The solvents were concentrated. The residue was purified by reverse phase HPLC (C18, 30- 80 % acetonitrile in H20 with 0.1 % formic acid) to afford the title compound (25 mg, 32% yield) as a white powder. Ή NMR (400 MHz, DMSO-d6) δ 8.86 (s, 1 H), 8.27 (s, 1 H), 8.05 (s, 1 H), 7.78 (s, 1 H), 7.68 (s, 1 H), 7.47 (s, 1 H), 7.09 (t, J = 54.4 Hz, 1 H), 3.93 (s, 3H), 3.45 (s, 2H), 2.87 - 2.56 (m, 2H), 2.11 - 1 .99 (m, 2H), 1 .98 - 1.75 (m, 2H). LCMS (ESI) m/z: 412.4 (M + 1)+.
[001298] Example 115
[001299] 10l-(Difluoromethyl)-2'-oxo-9l-(pyridin-3-yl)-2l.7l- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000268_0001
[001300] Step 1 : Ethyl 1 O'-formyl^'-oxo-g'-foyridin-S-vn^'J1- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000268_0002
[001301] To a solution of ethyl 10'-formyl-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (190 mg, 0.39 mmol) in 1 ,4-dioxane (8 mL) was added 3-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)pyridine (96 mg, 0.47 mmol), Pd(PPh3)4 (45 mg, 0.04mmol) and Na2C03 (124 mg, 1 .17 mmol) under N2 atmosphere. The reaction mixture was heated at 90 °C for overnight. After cooling to rt, the mixture was filtered, washed with DCM (30 mL). The filtrated was concentrated and the residue was purified by prep-TLC using MeOH in DCM (1/15) to afford the title compound (85 mg, 52% yield). LCMS (ESI) m/z: 415.7 (M + 1)+.
[001302] Step 2: Ethyl 10l-(difluoromethyl)-2'-oxo-9l-(pyridin-3-yl)-2l.7l- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000269_0001
[001303] To a solution of ethyl 10'-formyl-2'-oxo-9'-(pyridin-3-yl)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (85 mg, 0.21 mmol) in DCM (4 mL) was added dropwise DAST (169 mg, 1 .05 mmol) at 0 °C under N2 atmosphere. The reaction mixture was stirred at rt for overnight. The mixture was diluted with DCM (30 mL), washed with sat.NaHC03 solution (20 mL) and brine (20 mL). The solvent was concentrated to afford the title compound (30 mg, 33% yield). LCMS (ESI) m/z: 440.6 (M + 1)+.
[001304] Step 3: l O'^DifluoromethvD^'-oxo-g'^pyridin-S-vD^'J1- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000269_0002
[001305] To a solution of ethyl 10'-(difluoromethyl)-2'-oxo-9'-(pyridin-3-yl)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate ( 20 mg, 0.05 mmol) in EtOH (6 mL) was added NaOH (9 mg, 0.23 mmol) in H20 (1 .5 mL). The reaction mixture was stirred at rt for 2 h. The solvent was concentrated. The mixture was acidified to pH~1 by 1 N HCI and extracted with DCM (10 mL X 3). The combined organic layers were washed with brine (30 mL) and dried over Na2S04. The solvents were concentrated. The residue was purified by prep, reverse phase HPLC (C18, 30- 60 % acetonitrile in H20 with 0.1 % formic acid) to afford the title compound (1 .5 mg, 8% yield) as a white powder. 1H NMR (400 MHz, DMSO-de) δ 8.88 (s, 1 H), 8.68 (d, J = 4.8, 1 H), 8.65 (s, 1 H), 8.35 (s, 1 H), 7.88 (d, J = 7.9 Hz, 1 H), 7.61 - 7.51 (m, 2H), 6.96 (t, J = 54.4Hz, 1 H), 3.48 (s, 2H), 2.79 - 2.58 (m, 2H), 2.16 - 2.00 (m, 2H), 2.00 - 1 .82 (m, 2H). LCMS (ESI) m/z: 408.4 (M + 1)+.
[001306] Example 116
[001307] 10-(Difluoromethyl)-6-isopropyl-6-methyl-9-(oxetan-3-ylmethoxy)-2- oxo-6, 7-dihvdro-2H-pyrido[2, 1 -alisoquinoline-3-carboxylic acid
Figure imgf000270_0001
[001308] Step 1 : Ethyl 10-formyl-6-isopropyl-6-methyl-9-(oxetan-3-ylmethoxy)- -6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000270_0002
[001309] To a mixture of ethyl 10-formyl-9-hydroxy-6-isopropyl-6-methyl-2- oxo-6, 7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (640 mg,1 .73 mmol) in DMF (20 ml) were added K2C03 (716 mg, 5.19 mmol) and oxetan-3-ylmethyl 4- methylbenzenesulfonate (840 mg, 3.47 mmol). After stirring at 90 °C for 2hr, the reaction mixture was quenched with H20 and extracted with EtOAc and the organic layer was dried over Na2S04. Solvent was removed under vacuum and the residue was purified to provide ethyl 10-formyl-6-isopropyl-6-methyl-9-(oxetan-3-ylmethoxy)-2-oxo-6, 7-dihydro-2H- pyrido[2,1 -a]isoquinoline-3-carboxylate (450 mg,59% yield) as a yellow solid. LCMS (ESI) m/z: 440.3 (M + 1)+.
[001310] Step 2: Ethyl 10-(difluoromethyl)-6-isopropyl-6-methyl-9-(oxetan-3- ylmethoxy)-2-oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000270_0003
[001311] At 0 °C, to a mixture of ethyl 10-formyl-6-isopropyl-6-methyl-9-
(oxetan-3-ylmethoxy)-2-oxo-6, 7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (450 mg,1 .02 mmol) in DCM (20 ml) were added N,N-diethyl-1 ,1 ,1 -trifluoro-l4-sulfanamine (1 .320 mg, 8.80 mmol) and the reaction was stirred at 0 °C to room temperature overnight. The mixture was cooled to 0 °C, and extracted with NaHC03, water and DCM (20 ml_*3), the combined organic layers were washed with brine and dried over Na2S04. Solvents were removed under vacuum and the residue was purified by prep, reverse phase HPLC (C18, 0- 90% acetonitrile in H20 with 0.1 % formic acid) to provide ethyl 10-(difluoromethyl)-6- isopropyl-6-methyl-9-(oxetan-3-ylmethoxy)-2-oxo-6, 7-dihydro-2H-pyrido[2,1 -a]isoquinoline- 3-carboxylate (1 12 mg, 23.7% yield) as a white solid. LCMS (ESI) m/z: 462.7 (M + 1)+.
[001312] Step 3: 10-(Difluoromethyl)-6-isopropyl-6-methyl-9-(oxetan-3-
Figure imgf000271_0001
[001313] At r.t, to a mixture of ethyl 10-(difluoromethyl)-6-isopropyl-6-methyl- 9-(oxetan-3-ylmethoxy)-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (1 12 mg, 0.24 mmol) in THF (3 mL) were added LiOH (29.2 mg, 1 .22 mmol) and H20(1 mL). The reaction was stirred at r.t for 1 hr. Solvents were removed, the mixture was poured into water (20mL) and the PH adjusted to 6 with 5% HCI (2mL). The solid was filtered and the filter cake was washed by water, dried under vacuum to give the 10-(difluoromethyl)-6-isopropyl- 6-methyl-9-(oxetan-3-ylmethoxy)-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3- carboxylic acid (85 mg, 80% yield) as a white solid. LCMS (ESI) m/z: 434.2 (M + 1)+. 1H NMR (400 MHz, DMSO-d6) δ 8.57(s, 1 H), 8.16 (s 1 H), 7.42 (s, 1 H), 7.33 (s, 1 H), 7.21 (s, 1 H), 7.07(t, J=54Hz, 1 H), 4.72 (dd, J = 7.6, 6.3 Hz, 2H), 4.47 (td, J=6.0, 3.2 Hz, 2H), 4.39 (dt, J = 16.3, 9.9 Hz, 2H), 3.46-3.40 (m, 1 H), 3.27(s, 2H), 1 .78-1 .74 (m, 1 H), 1 .69(s, 3H), 0.82 (d, J = 6.8 Hz, 3H), 0.64 (d, J = 6.8 Hz, 3H).
[001314] Example 117
[001315] 10-Cvclopropyl-6-isopropyl-9-methoxy-6-methyl-2-oxo-6,7-dihydro- 2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000271_0002
[001316] Step 1 : Ethyl 9-(benzyloxy)-10-cvclopropyl-6-isopropyl-6-methyl-2- oxo-6,7-dihvdro-2H-pyr -alisoquinoline-3-carboxylate
Figure imgf000271_0003
[001317] Under a stream of nitrogen ethyl 9-(benzyloxy)-10-bromo-6- isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (50 mg, 0.10 mmol), cyclopropylboronic acid (25.8 mg, 0.30 mmol), Pd(Ph3)4 (10.3 mg, 0.01 mmol), Cs2C03 (97.7 mg, 0.30 mmol), PhMe-H20 (1 mL - 1 drop) were placed into a microwave tube, then the mixture was stirred at 1 10 °C for 3hr. The reaction mixture was added to water 20 mL, and extracted with EtOAc. The organic layer was dried over Na2S04. Solvents were removed under vacuum and the residue was purified to provide ethyl 9-(benzyloxy)-10-cyclopropyl-6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 - a]isoquinoline-3-carboxylate (40 mg, 86% yield) as an oily liquid (crude). LCMS (ESI) m/z: 472.2 (M + 1)+.
[001318] Step 2: Ethyl 10-cvclopropyl-9-hvdroxy-6-isopropyl-6-methyl-2-oxo- 6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000272_0001
[001319] A solution of ethyl 9-(benzyloxy)-10-cyclopropyl-6-isopropyl-6- methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (40 mg, 0.08 mmol) in TFA (1 mL) was heated to 70 °C for 1 hr. The mixture was extracted with EtOAc. The organic layer was dried over Na2S04. Solvents were removed under vacuum and the residue was purified to provide ethyl 10-cyclopropyl-9-hydroxy-6-isopropyl-6-methyl-2-oxo-6,7-dihydro- 2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (20 mg, 62% yield) as a white solid. LCMS (ESI) m/z: 382.3 (M + 1)+ .
[001320] Step 3: 10-Cvclopropyl-6-isopropyl-9-methoxy-6-methyl-2-oxo-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000272_0002
[001321] To a solution of NaH (3.8 mg, 0.16 mmol) in DMF (1 mL) was added ethyl 10-cyclopropyl-9-hydroxy-6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 - a]isoquinoline-3-carboxylate (20 mg, 0.05 mmol) at r.t for 20 min. and Mel (22.3 mg, 0.157 mmol) was added in one portion at r.t for 2 hr. The reaction was extracted with EtOAc. The organic layer was dried over Na2S04. Solvents were removed under vacuum and the residue was purified by reverse phase HPLC (C18, 0-40% acetonitrile in H20 with 0.1 % formic acid) to provide 10-cyclopropyl-6-isopropyl-9-methoxy-6-methyl-2-oxo-6,7-dihydro- 2H-pyrido[2,1 -a]isoquinoline-3-carboxylic acid (5 mg, 26% yield) as a white solid. LCMS (ESI) m/z: 368.3 (M + 1)+ . Ή NMR (400 MHz, DMSO-d6) δ 8.53(s, 1 H), 7.44 (s 1 H), 7.40 (s, 1 H), 7.03(s, 1 H), 3.91 (s, 3H), 3.23 (d, J=3.8Hz, 1 H), 3.17(s, 1 H), 2.12-2.05 (m, 1 H), 1 .82- 1 .75 (m, 1 H), 1 .66 (s, 3H), 0.95-0.83(m, J=13.0, 6.3 Hz, 4H), 0.81 (d, J=6.8 Hz, 3H), 0.61 (d, J = 6.8 Hz, 3H).
[001322] Example 118
[001323] 10-(Difluoromethyl)-6-isopropyl-6-methyl-9-(1 -methyl- 1 H-pyrazol-4- yl)-2-oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000273_0001
[001324] Step 1 : Ethyl 10-formyl-6-isopropyl-6-methyl-9-(1 -methyl- 1 H-pyrazol- 4-yl)-2-oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000273_0002
[001325] Under a stream of nitrogen, ethyl 10-formyl-6-isopropyl-6-methyl-2- oxo-9-(((trifluoromethyl)sulfonyl)oxy)-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (400 mg, 0.8 mmol), 1 -methyl-4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)-1 H-pyrazole (200 mg, 0.96 mmol), Pd(PPh3)4 (96 mg, 0.08 mmol), Cs2C03 (522 mg, 1 .6 mmol) and toluene (40 ml_) were placed into a single round bottom flask, then stirred for 3 h at 100 °C. After the reaction, the reaction mixture was added to water (100 ml_), and extracted with ethyl acetate, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The oil was dissolved in a small amount of ethyl acetate, packed to silica gel chromatography, subjected to column purification using hexane/ethyl acetate as an eluent. The solvent of the obtained solution can be distilled off under reduced pressure to afford the title compound (173 mg, 50% yield). LCMS (ESI) m/z: 434.2 (M + 1)+.
[001326] Step 2: Ethyl 10-(difluoromethyl)-6-isopropyl-6-methyl-9-(1 -methyl- 1 H-pyrazol-4-yl)-2-oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylat
Figure imgf000274_0001
[001327] To a solution of ethyl 10-formyl-6-isopropyl-6-methyl-9-(1 -methyl-1 H- pyrazol-4-yl)-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (173 mg, 0.4 mmol) in DCM (3 mL) was added a solution of DAST (322 mg, 2 mmol) in DCM (2 mL) dropwise over 30 min at 0 °C. After the reaction mixture was finished, the mixture was poured into saturated NaHC03 aqueous at 0 °C. The new mixture was extracted with DCM for twice. The combined DCM layers were washed with brine, dried over MgS04 and concentrated to afford the title compound (83 mg, 45.6% yield). LCMS (ESI) m/z: 456.2 (M +
[001328] Step 3: 10-(Difluoromethyl)-6-isopropyl-6-methyl-9-(1 -methyl- 1 H-
Figure imgf000274_0002
[001329] To a solution of ethyl 10-(difluoromethyl)-6-isopropyl-6-methyl-9-(1 - methyl-1 H-pyrazol-4-yl)-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (200 mg, 0.44 mmol) in EtOH (50 mL) was added NaOH (88 mg, 2.2 mmol) at room temperature After stirring at room temperature for 2 h, the reaction mixture was acidified to pH~1 and extracted with EtOAc three times. The combined organic layers were washed with brine and dried over Na2S04. Solvent were removed and the residue was purified by reverse phase HPLC (C18 0-40% MeCN in H20 with 0.1 % formic acid) to afford the title compound (80 mg, 42% yield) as a white solid. Ή NMR (400 MHz, DMSO-d6) δ 8.60 (s, 1 H), 8.30 (s, 1 H), 7.77 (s, 1 H), 7.65 (s, 1 H), 7.64-7.59 (m, 1 H), 7.23 (s, 0.25H), 7.09 (s, 0.50H), 6.96 (s, 0.25H), 3.93 (s, 3H), 3.35 (s, 2H), 1 .79-1 .74 (m, 1 H), 1 .71 (s, 3H), 0.82 (d, J = 6.8 Hz, 3H), 0.65 (d, J = 6.8 Hz, 3H). LCMS (ESI) m/z: 428.18 (M + 1 )+.
[001330] Example 119
[001331] 10-(Difluoromethyl)-6-isopropyl-6-methyl-2-oxo-9-propyl-6,7-dihydro- 2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000275_0001
[001332] Step 1 : Ethyl 9-allyl-10-formyl-6-isopropyl-6-methyl-2-oxo-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000275_0002
[001333] Under a stream of nitrogen, ethyl 10-formyl-6-isopropyl-6-methyl-2- oxo-9-(((trifluoromethyl)sulfonyl)oxy)-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (400 mg, 0.8 mmol), 2-allyl-4,4,5,5-tetramethyl-1 ,3,2-dioxaborolane (269 mg, 1 .6 mmol), Pd(PPh3)4 (96 mg, 0.08 mmol), Cs2C03 (522 mg, 1 .6 mmol) and toluene (40 mL) were placed into a single round bottom flask, then stirred for 3 h at 100 °C. After the reaction, the reaction mixture was added to water (100 mL), and extracted with ethyl acetate. The organics were dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure, an oil liquid was obtained. This was dissolved in a small amount of ethyl acetate, packed to silica gel chromatography, and subjected to column purification using hexane/ethyl acetate as an eluent. The solvent of the obtained solution can be distilled off under reduced pressure to afford the title compound (170 mg, 54% yield). LCMS (ESI) m/z: 394.2 (M + 1)+.
[001334] Step 2: Ethyl 10-formyl-6-isopropyl-6-methyl-2-oxo-9-propyl-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000275_0003
[001335] A mixture of ethyl 9-allyl-10-formyl-6-isopropyl-6-methyl-2-oxo-6,7- dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (300 mg, 0.76 mmol) and Pd-C (81 mg, 0.68 mmol) in EtOH (10 mL) was stirred at room temperature under a stream of hydrogen overnight. The resulting mixture was filtered through a pad of Celite and the filtered was concentrated under reduced pressure to give the crude product which was purified by flash chromatography (silica gel, 0-50% EtOAc in petroleum ether) to afford the title compound (241 mg, 80% yield). LCMS (ESI) m/z: 396.4 (M + 1 )+.
[001336] Step 3: Ethyl 10-(difluoromethyl)-6-isopropyl-6-methyl-2-oxo-9- propyl-6,7-dihvdro-2H-p -alisoquinoline-3-carboxylate
Figure imgf000276_0001
[001337] To a solution of ethyl 10-formyl-6-isopropyl-6-methyl-2-oxo-9-propyl- 6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (241 mg, 0.6 mmol) in DCM (3 mL) was added a solution of DAST (483 mg, 3.0 mmol) in DCM (2 mL) by dropwise over 30 min at 0 °C. After the reaction mixture was finished, the mixture was poured into saturated NaHC03 aqueous at 0 °C. The new mixture was extracted with DCM. The combined DCM layers were washed with brine, dried over MgS04 and concentrated to afford the title compound (140 mg, 56% yield). LCMS (ESI) m/z :418.3 (M + 1)+.
[001338] Step 4: 1 10-(Difluoromethyl)-6-isopropyl-6-methyl-2-oxo-9-propyl- 6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000276_0002
[001339] To a solution of ethyl 10-(difluoromethyl)-6-isopropyl-6-methyl-2-oxo- 9-propyl-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (140 mg, 0.34 mmol) in EtOH (50 mL) was added NaOH (67 mg, 1 .68 mmol) at room temperature After stirring at room temperature for 2 h, the reaction mixture was acidified to pH~1 and extracted with EtOAc three times. The combined organic layers were washed with brine and dried over Na2S04. Solvent was removed and the residue was purified by reverse phase HPLC (C18 0-40% MeCN in H20 with 0.1 % formic acid) to afford the title compound (18 mg, 13.6% yield). LCMS (ESI) m/z: 390.2 (M + 1 )+.1 H NMR (400 MHz, DMSO-d6) δ 8.59 (s, 1 H), 8.19 (s, 1 H), 7.46 (d, J = 5.7 Hz, 2H), 7.32 (s, 0.25H), 7.18 (s, 0.5H), 7.05 (s, 0.25H), 3.31 (s,2H), 2.78 (t, J = 7.7 Hz, 2H), 1 .76 - 1 .71 (m,1 H), 1 .69 (s, 3H), 1 .64 - 1 .57 (m, 2H), 0.95 (t, J = 7.3 Hz, 3H), 0.79 (d, J = 6.8 Hz, 3H), 0.63 (d, J = 6.8 Hz, 3H).
[001340] Example 120 [001341] 9'-(Oxetan-3-ylmethoxy)-2'-oxo-10'-(trifluoromethylV
2',7'dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000277_0001
[001342] Step 1 : 3-(Benzyloxy)-4-(trifluoromethyl)benzoic acid
Figure imgf000277_0002
[001343] To a mixture of 3-fluoro-4-(trifluoromethyl)benzoic acid (15 g,
72.1 mmol) and phenylmethanol (12.44 g,1 15.2mmol) in DMSO (150ml) was added NaH (4.14 g, 173.04mmol) dropwise at 0°Cand stirred at R.T for 12 h. H20 (0.1 L) was added to quench the reaction, and added HCI (cone). Filter white solid and obtained mixture was extracted with EtOAc (2 x0.2 L). The combined organic layers were washed with brine, dried over MgS04 and concentrated under reduced pressure. The compound was purified by silica gel column purification (PE:EA=10:1 ) to give 18.6 g . LCMS (ESI) m/z: 295.2 (M-1 )".
[001344] xy)-4-(trifluoromethyl)phenyl)methanol
Figure imgf000277_0003
[001345] To a mixture of 3-(benzyloxy)-4-(trifluoromethyl)benzoic acid (18.6 g,
65.9mmol) in THF (150ml) was added BH3 (132ml, 131 .8mmol) dropwise at 0 °Cand stirred at R.T for 12 h. H20 (0.15 L) was added to quench the reaction at 0 °C, and the mixture was concentrated under reduced pressure, diluted with water and extracted with EtOAc (2 x0.2 L). The combined organic layers were washed with brine, dried over MgS04 and concentrated under reduced pressure. Silica gel column purification (PE:EA=10:1) gave 16 g of solids (90.3% yield). LCMS (ESI) m/z: 283.4 (M + 1)+.
[001346]
Figure imgf000277_0004
[001347] A solution of PPh3 (21 g, 0.08 mol) in THF (160 ml) was added dropwise in the mixture of (3-(benzyloxy)-4-(trifluoromethyl)phenyl)methanol (16 g, 0.057 mol) and CBr4 (26.5 g,0.08mol) in THF (160 ml) at 0 °C. The resulting mixture was stirred at rt for 3 h before it was diluted with Et20 (600 ml). The obtained suspension was filtered through a pad of celite, and the filtrate was concentrated to give the crude product which was purified by flash column chromatography (silica gel, 0-6% CH2CI2 in petroleum ether) to afford the title compound (14 g, 71 .8% yield). Ή NMR (400 MHz, CDCI3) δ 7.57 (d, J = 7.9, 1 H), 745 (d ,J = 7.4 Hz, 2H), 7.42 - 7.36 (m, 2H), 7.34 - 7.29 (m, 1 H), 7.10 (s, 1 H), 6.97 (dd, J =7.9, 0.5 Hz, 1 H), 5.20 (s, 2H), 4.72 (s, 2H).
[001348] Step 4: Ethyl 1 -(3-(benzyloxy)-4-(trifluoromethyl)benzyl)cvclobutane-
1 -carboxylate
Figure imgf000278_0001
[001349] To a freshly prepared LDA solution in THF (2 N, 21 .75 ml_) was added the solution of ethyl cyclobutanecarboxylate (3.7 g, 0.029 mol) in THF (50 ml) dropwise at -78 °C. The resulting mixture was stirred at -78 °C for 1 .5 h before the solution of 2-(benzyloxy)-4-(bromomethyl)-1 -(trifluoromethyl)benzene (5.0 g, 0.015 mol) in THF (50 ml) was added dropwise. The reaction mixture was allowed to warm up to r.t slowly and stirred for 12 h. NH4CI solution (sat. aq., 60 ml) was added to quench the reaction, and the obtained mixture was extracted with EtOAc (4x200 ml). The combined organic layers was washed with 1 N HCI (100 ml), NaHC03 (sat. aq., 100 ml) and brine. The organic phase was dried over Na2S04, filtered and concentrated to give the crude product which was purified by flash column chromatography (silica gel, PE: EA=5:1) to afford the title compound (2.3 g, 40 % yield). LCMS (ESI) m/z: 393.6 (M + 1)+. 1H NMR (400 MHz, CDC ) δ 7.51 - 7.46 (m, 1 H),7.44 (d, J = 7.3 Hz, 2H), 7.37 (t, J= 7.4 Hz, 2H), 7.30 (t, J = 7.2 Hz, 1 H), 6.82 (s, 2H), 5.18 (s, 2 H), 4.1 1 (q, J = 7.1 Hz, 2H), 3.12 (s, 2H), 2.30 (td, J = 9.8, 4.8 Hz, 4H), 2.08 (dt, J = 12.5, 8.5 Hz, 2H), 1 .23 (t, J = 7.1 Hz, 3H) .
[001350] Step 5: 1 -(3-(Benzyloxy)-4-(trifluoromethyl)phenyl)cyclobutane-1 - carboxylic acid
Figure imgf000278_0002
[001351] To a solution of ethyl 1 -(3-(benzyloxy)-4-
(trifluoromethyl)benzyl)cyclobutane-1 -carboxylate (2.3 g, 5.8 mmol) in THF (20 ml), MeOH (10 ml) and water (10 ml) was added LiOH (0.84 g, 35.2 mmol). After stirred at r.t for 4 hr, the resulting mixture was neutralized with 1 N HCI and extracted with EtOAc (60 ml). The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to give the crude product which was purified by flash chromatography (silica gel, PE:EA=3:1) to afford the title compound (1 .7 g, 85% yield) as a white solid. 1H NMR (400 MHz, CDCI3) δ 7.49 - 7.29 (m, 6H), 6.85 - 6.72 (m, J = 6.3 Hz, 2H), 5.16 (s, 2H), 3.09 (s, 2H), 2.46 - 2.36 (m, 2H), 2.05 - 1 .99 (m, 2H), 1 .94 - 1 .87 (m, 2H).
[001352] Step 6: 1 -(3-(Benzyloxy)-4-(trifluoromethyl)benzyl)cyclobutan-1 - amine
Figure imgf000279_0001
[001353] To a solution of 1 -(3-(benzyloxy)-4- (trifluoromethyl)phenyl)cyclobutane-1 -carboxylic acid (1 .7 g,4.8 mmol) in toluene (20 ml) was added TEA (0.7 g,7.0 mmol), followed by DPPA (2.0 g, 7.0 mmol). The mixture was stirred at 60°C for 2 h. After the reaction was finished, the mixture was cooled to room temperature and added with concentrated HCI (6 ml). The new mixture was heated to 60°C for 2 h. After the reaction was finished, the mixture was cooled to 0°C and basified by 1 N NaOH aqueous. The organic layer was washed with brine, dried over MgS04. filtered and concentrated to give the crude product which was purified by flash chromatography (silica gel, DCM: MeOH=20:1) to afford the title compound (1 .0 g, 62.5% yield) as a white solid. LCMS (ESI) m/z: 336.2 (M + 1)+.
[001354] Step 7: N-(1 -(3-(Benzyloxy)-4
(trifluoromethvDbenzvDcyclobutvDformamide
Figure imgf000279_0002
[001355] To solution of 1 -(3-(benzyloxy)-4-(trifluoromethyl) benzyl)cyclobutan- 1 -amine (1 .0g, 3mmol) in toluene was acetic formic anhydride (10ml) by dropwise at room temperature The mixture was stirred for 30 min at room temperature. After the reaction was finished, the mixture was concentrated under vacuo and used to next step without purification. LCMS (ESI) m/z: 364.5 (M + 1)+.
[001356] Step 8: 6'-(Benzyloxy)-7'-(trifluoromethyl)-4'H-spiro[cyclobutane-1 ,3'- isoquinolinel
Figure imgf000279_0003
[001357] The residue of N-(1 -(3-(benzyloxy)-4- (trifluoromethyl)benzyl)cyclobutyl)formamide (3 mmol) was dissolved into MeCN (20 ml). The mixture was added with POCI3 (0.54 g, 3.5 mmol) dropwise over 30 min and stirred for 5h at room temperature. After the reaction was finished, the reaction mixture was cooled to 0°C and basified by Ammonium hydroxide to pH 9-10. The new mixture was extracted with DCM for twice. The combined organic layers were washed with brine, and concentrated. The crude product (1 .0g, yield: 100%) was used to next step without purification. LCMS (ESI) m/z: 346.24. (M + 1)+.
[001358] Step 9: Ethyl 9l-n3enzyloxyV2l-oxo-10l-arifluoromethylV1 '.2l.7l.1 1 bl- tetrahydrospirofcyclobu '-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000280_0001
[001359] The residue of 6'-(benzyloxy)-7'-(trifluoromethyl)-4'H- spiro[cyclobutane-1 ,3'-isoquinoline] (1 .0 g, 3 mmol) was dissolved in EtOH (20 ml) and added with ethyl 2-(ethoxymethylene)-3-oxobutanoate (1 .5 g, 8 mmol). The mixture was heated to 85°C overnight. After the reaction was finished, the mixture was cooled to 0 °C and stirred for 2 h. The solid was filtered and washed with MTBE to afford title compound (1 .0 g, yield: 67%). LCMS (ESI) m/z: 486.3 (M + 1)+.
[001360] Step 10: Ethyl 9'-(benzyloxy)-2'-oxo-10'-(trifluoromethyr)-2'.7'- dihydrospirofcyclobutan '-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000280_0002
[001361] To the suspension of ethyl 9'-(benzyloxy)-2'-oxo-10'-(trifluoromethyl)- 1 ',2',7',1 1 b'-tetrahydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (0.2 g, 0.4 mmol) in DME (5 ml) was added Tetrachloro-p-benzoquinone (83.6 mg, 0.34 mmol) at room temperature. The mixture was heated to 75°C overnight. After the reaction was finished, the solid was filtered and washed with DME to afford title compound (150 mg, yield: 77.7%) as a white solid. LCMS (ESI) m/z: 484.5 (M + 1 )+.
[001362] Step 1 1 : Ethyl 9'-Ι^Γθχν-2'-οχο-10'-grifluoromethvD^'J'- dihydrospirofcyclobutan '-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000280_0003
[001363] A solution of ethyl 9'-(benzyloxy)-2'-oxo-10'-(trifluoromethyl)-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (150 mg, 0.3 mmol) in TFA (4 ml) was stirred at 65 °C. After the reaction was finished, the mixture was concentrated. The residue was diluted with water and extracted DCM, washed with K2C03 solution, dried over MgSC and concentrated. The crude was purified with flash chromatography to afford the title compound (70 mg, 63.6% yield). LCMS (ESI) m/z: 394.4 (M + 1 )+.
[001364] Step 12: Ethyl 9'-(oxetan-3-ylmethoxyV2'-oxo-10'-ftrifluoromethvn-
2',7'-dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000281_0001
[001365] A Schlenk flask was charged with ethyl 9'-hydroxy-2'-oxo-10'- (trifluoromethyl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (70 mg, 0.2mmol), oxetan-3-ylmethyl 4-methylbenzenesulfonate (73.5 mg, 0.54 mmol) and DMF (2 ml). To this reaction mixture, K2C03 (1 10 mg, 0.8 mmol) was added and then stirred for 6 h by heating at 80 °C. After cooling, the obtained inorganic salt was filtered off. Ethyl acetate was added to the organic layer and washed with 1 % brine. After drying with anhydrous magnesium sulfate, the solvent was evaporated, purified by column
chromatography to afford the title compound (65 mg, 70.6% yield). LCMS (ESI) m/z: 464.2 (M + 1 )+.
[001366] Step 13: 9'-(Oxetan-3-ylmethoxy)-2'-oxo-10'-(trifluoromethyl)-2'.7'- dihydrospirofcyclobutane-1 '-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000281_0002
[001367] To a solution of ethyl 9'-(oxetan-3-ylmethoxy)-2'-oxo-10'- (trifluoromethyl)-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (65 mg, 0.14 mmol) in MeOH (2 ml) and water (0.5 ml) was added LiOH (20.16 mg, 0.84 mmol) at room temperature The reaction mixture was stirred at room temperature for 1 h, and detected with LCMS. After the completion of the reaction, the mixture was acidified to pH~4 and purified by reverse phase HPLC (C18 0-40% MeCN in H20 with 0.1 % formic acid) to afford the title compound (22 mg, 36.7% yield) as a white solid. LCMS (ESI) m/z: 436.2 (M + 1 )+. Ή NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1 H), 8.24 (s, 1 H), 7.55-7.41 (m,
J=9.7Hz,2H), 4.71 (dd, J=7.9,6.0Hz,2H), 4.57-4.36 (m, J = 12, 6.1 Hz, 4H), 3.51 -3.42 (m, J = 9.6 Hz,3H),2.75-2.62 (m, J =1 1 .3 Hz, 2H), 2.05 - 1 .85 (m, 4H).
[001368] Example 121
[001369] 10'-chloro-9'-(3-methoxypropoxy)-3.3-dimethyl-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000282_0001
[001370] Step 1 : 4-Chloro-3-(3-methoxypropoxy)benzaldehvde
Figure imgf000282_0002
[001371] To a solution of 4-chloro-3-hydroxybenzaldehyde (10.39 g, 66.4 mmol) in Ν,Ν-dimethylformamide (DMF) (100 mL) was added potassium carbonate (45.9 g, 332 mmol) and 1 -bromo-3-methoxypropane (12.19 g, 80 mmol). The reaction was stirred at room temp overnight. The suspension was filtered, washed with ethyl ether (2 x 50 mL) and the combined solution washed with water (100 mL). The aqueous layer washed with ethyl ether (2 x 50 mL). The combined organic layers were washed with LiCI (aq 5%, 50 mL), washed with brine, dried with sodium sulfate, and concentrated to give 4-chloro-3-(3- methoxypropoxy)benzaldehyde (12.52 g, 54.8 mmol, 83 % yield) as a yellow oil. LCMS (ESI) m/z: 229.2 (M + 1)+.
[001372] Step 2: (4-Chloro-3-(3-methoxypropoxy)phenyl)methanol
Figure imgf000282_0003
[001373] To a solution of 4-chloro-3-(3-methoxypropoxy)benzaldehyde (12.52 g, 54.8 mmol) in methanol (200 mL) at RT was added sodium borohydride (4.14 g, 1 10 mmol) portion wise and the mixture stirred for 1 hr. The reaction was poured into half sat NH4CI (aq. 150 mL) and filtered. The filtration was slow, so the remaining slurry (~1/2 filtered) was treated with Et20 (200 mL) for two solution phases. The organics were dried (brine, Na2S04) and concentrated to give (4-chloro-3-(3-methoxypropoxy)phenyl)methanol (12.3 g, 53.3 mmol, 97 % yield) as a yellow oil. Ή NMR (400 MHz, DMSO-d6) δ 7.35 (m, 1 H), 7.02 (s, 1 H), 6.93 (m, 1 H), 5.21 (bs, 1 H), 4.45 (s, 2H), 4.05 (m, 2H), 3.41 (m, 2H), 3.20 (s, 4H), 1 .99 (m, 2H).
[001374] -(Bromomethyl)-1 -chloro-2-(3-methoxypropoxy)benzene
Figure imgf000283_0001
[001375] To a solution of (4-chloro-3-(3-methoxypropoxy)phenyl)methanol
(12.26 g, 53.1 mmol) in tetrahydrofuran (THF) (200 mL) at RT was treated with phosphorus tribromide (6.01 mL, 63.8 mmol) dropwise and stirred at ambient temperature. The mixture grew warm during addition. The reaction was complete after 15 min. The reaction was cooled to 0 °C, slowly poured onto a mixture of ice and sat NaHC03 (aq), and then extracted with Et20. The combined organics were dried (brine, Na2S04) and purified by silica chromatography (0-25% EtOAc in hexanes) to give 4-(bromomethyl)-1 -chloro-2-(3- methoxypropoxy)benzene (8.4 g, 28.6 mmol, 53.8 % yield) as an oil. 1 H NMR (400 MHz, DMSO-de) δ 7.39 (m, 1 H), 7.21 (s, 1 H), 7.01 (m, 1 H), 5.21 (bs, 1 H), 4.65 (s, 2H), 4.05 (m, 2H), 3.45 (m, 2H), 3.23 (s, 4H), 1 .95 (m, 2H).
[001376] Step 4: Methyl 1 -(4-chloro-3-(3-methoxypropoxy)benzyl)-3,3-
Figure imgf000283_0002
[001377] To LDA (17.03 mL, 17.03 mmol) (1 M in THF) at 0 °C was added methyl 3,3-dimethylcyclobutane-1 -carboxylate (1 .94 g, 13.6 mmol) and stirred for 5 min. A solution of 4-(bromomethyl)-1 -chloro-2-(3-methoxypropoxy)benzene (2 g, 6.81 mmol) in THF (5mL) was added dropwise and then the mixture allowed to warm to RT. The mixture was l uenched with ice and NH4CI (aq) and extracted with EtOAc. The organics were combined, dried (brine, Na2S04) and concentrated to an oil. The oil was purified on silica gel hex:EtOAc (eluting at ~10%) to give methyl 1 -(4-chloro-3-(3-methoxypropoxy)benzyl)-3,3- dimethylcyclobutane-1 -carboxylate (2.27 g, 6.40 mmol, 94 % yield) as a yellow oil. 1H NMR (400 MHz, DMSO d6) δ 7.25 (m, 1 H), 6.89 (s, 1 H), 6.61 (m, 1 H), 4.05 (m, 2H), 3.62 (s, 3H), 3.45 (m, 2H), 3.21 (m, 3H), 2.99 (s, 2H), 2.05 (m, 4H), 0.99 (s, 6H).
[001378] Step 5: 1 -(4-Chloro-3-(3-methoxypropoxy)benzyl)-3,3- dimethylcyclobutane carboxylic acid
Figure imgf000283_0003
[001379] To a solution of methyl 1 -(4-chloro-3-(3-methoxypropoxy)benzyl)- 3,3-dimethylcyclobutane-1 -carboxylate (2.27 g, 6.40 mmol) in THF:MeOH (10 mL : 20 mL) was added LiOH (1 .23 g, 51 .2 mmol) in water (10 mL). The mixture was warmed to 55 °C for 3 hrs., poured into 100 mL HCI (1 N aq), and extracted with EtOAc. The organic phase was dried (brine, Na2S04) and concentrated to an orange oil. The oil crystallized upon standing to give 1 -(4-chloro-3-(3-methoxypropoxy)benzyl)-3,3-dimethylcyclobutane-1 - carboxylic acid (1 .84 g, 5.40 mmol, 84 % yield). LCMS (ESI) m/z: 339.4 (M-1)".
[001380] Step 5: N-(1 -(4-Chloro-3-(3-methoxypropoxy1benzyr)-3.3- dimethylcyclobutvDformamide
Figure imgf000284_0001
[001381] To 1 -(4-chloro-3-(3-methoxypropoxy)benzyl)-3,3- dimethylcyclobutane-1 -carboxylic acid (1 .84 g, 5.40 mmol) was added TEA (0.903 mL, 6.48 mmol), tert-butanol (10 mL) and DPPA (1 .396 mL, 6.48 mmol). The reaction was heated to 80 °C under nitrogen. Full conversion to the isocyanate was observed after 30 min. After 1 hr. the mixture was poured into 1 N HCI (aq) at 80 °C. A precipitate formed. Dioxane (10 mL) was added to dissolve solids and the mixture stirred for 30 minutes, cooled, poured into NaHC03 (sat aq), extracted with EtOAc, dried (brine, Na2S04), and concentrated to an off- white solid. The solid was then dissolved in 4-dioxane (40 mL) before a pre-mixed solution of formic acetic mixed anhydride (6.8M in formic acid) 2 mL was added. The suspension was heated until all solids were dissolved and then the mixture cooled to RT. Another 10 mL of anhydride was added and the mixture, heated to 80 °C for one hour. The mixture was concentrated to a yellow oil and used directly in the next reaction. LCMS (ESI) m/z: 340.4 (M + 1)+.
[001382] Step 7: 7'-Chloro-6'-(3-methoxypropoxy)-3.3-dimethyl-4'H-
Figure imgf000284_0002
[001383] To N-(1 -(4-chloro-3-(3-methoxypropoxy)benzyl)-3,3- dimethylcyclobutyl)formamide (1 .83 g, 5.38 mmol) in acetonitrile (50 mL) was added POCI3 (0.502 mL, 5.38 mmol) and the mixture heated to 80 °C for 1 h. The reaction was added to ice and partitioned between NaHC03 (aq) and EtOAc. The aqueous phase was removed, washed again with EtOAc. The organics were combined, dried (brine, Na2S04) and concentrated to give 7'-chloro-6'-(3-methoxypropoxy)-3,3-dimethyl-4'H-spiro[cyclobutane- 1 ,3'-isoquinoline] (1 .733 g, 5.38 mmol, 100 % yield) as a red oil. The material was used directly in the subsequent reaction. LCMS (ESI) m/z: 322.3 (M + 1)+.
[001384] Step 8: Ethyl 10'-chloro-9'-(3-methoxypropoxy)-3,3-dimethyl-2'-oxo- 1 ',2',7',1 1 b'-tetrahvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000285_0001
[001385] A solution of 7'-chloro-6'-(3-methoxypropoxy)-3,3-dimethyl-4'H- spiro[cyclobutane-1 ,3'-isoquinoline] (1 .73 g, 5.38 mmol) and ethyl 2-(ethoxymethylene)-3- oxobutanoate (3.75 ml, 21 .5 mmol) in ethanol was heated to 80 °C for 16 hr. The mixture was cooled, partitioned between water and EtOAc, and extracted with additional EtOAc. The organic layers were dried (brine, Na2S04) and concentrated to give crude ethyl 10'- chloro-9'-(3-methoxypropoxy)-3,3-dimethyl-2'-oxo-1 ',2',7',1 1 b'-tetrahydrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate as a red oil. LCMS (ESI) m/z: 462.3 (M + 1)+.
[001386] Step 9: Ethyl 10'-chloro-9'-(3-methoxypropoxy)-3,3-dimethyl-2'-oxo- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000285_0002
[001387] A solution of ethyl 10'-chloro-9'-(3-methoxypropoxy)-3,3-dimethyl-2'- oxo-1 ',2', 7', 1 1 b'-tetrahydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (2.488 g, 5.39 mmol) and DDQ (3.06 g, 13.46 mmol) in 1 ,2-dimethoxyethane (DME) (40 mL) was heated to 80 °C for 3 hr. The mixture was partitioned between NaHC03 (Aq) and EtOAc. The organics were concentrated and the residue purified on silica (0-10% MeOH in DCM) twice to give 10 ethyl 10'-chloro-9'-(3-methoxypropoxy)-3,3-dimethyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (400 mg, 0.870 mmol, 16.15 % yield) as a light brown solid. LCMS (ESI) m/z: 460.4 (M + 1)+.
[001388] Step 1 1 : 10'-Chloro-9'-(3-methoxypropoxy)-3.3-dimethyl-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000286_0001
[001389] A solution of ethyl 10'-chloro-9'-(3-methoxypropoxy)-3,3-dimethyl-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (100 mg, 0.217 mmol) in methanol (10 ml_) and 1 ,4-dioxane (5.00 ml_) was treated with LiOH (62.5 mg, 2.61 mmol) in water (5.00 ml_) and heated to 60 °C for 1 hr. The solution was concentrated and partitioned between HCI (1 N) and EtOAc. The organic phase was dried (Na2S04, brine) and purified by reverse phase HPLC to give 10'-chloro-9'-(3- methoxypropoxy)-3,3-dimethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid (35.8 mg, 0.083 mmol, 38.1 % yield) as a light tan solid. 1 H NMR (400 MHz, c/6-DMSO-d6) δ ppm 8.84 (s, 1 H), 8.15 (s, 1 H), 7.38 (s, 1 H), 7.34 (s, 1 H), 4.21 (t, J= 6.2 Hz 2 H), 3.51 (t, J = 6.2 Hz, 2H), 3.36 (s, 2H), 3.21 (s, 3 H), 2.45 (m, 2H), 2.08 - 1 .98 (m, 4 H), 1 .24 (s, 3H), 1 .17 (s, 3H). LCMS(ESI+)(m/z):432.7 (M + 1)+.
[001390] Example 122
[001391] 10'-chloro-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihydrospirofcyclopentane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000286_0002
[001392] Step 1 : Methyl 1 -(4-chloro
methoxypropoxy)benzyl)cvclopentane
[001393] carboxylate
Figure imgf000286_0003
[001394] To LDA (17.03 ml_, 17.03 mmol) (1 M in THF) at 0 °C was added methyl cyclopentanecarboxylate (1 .75 g, 13.6 mmol) and the mixture stirred for 5 min. A solution of 4-(bromomethyl)-1 -chloro-2-(3-methoxypropoxy)benzene (2 g, 6.8 mmol) in THF (5ml_) was added dropwise then the mixture was allow to warm to RT. The mixture was quenched with ice and NH4CI (aq), and then extracted with EtOAc. The organics were combined, dried (brine, Na2S04), and concentrated to an oil. The oil was purified on silica gel hex:EtOAc (eluting at -10%) to give methyl 1 -(4-chloro-3-(3- methoxypropoxy)benzyl)cyclopentane-1 -carboxylate (2.28 g, 6.69 mmol, 98 % yield) as a yellow oil. LCMS (ESI) (m/z): 363.3 (M+23)+.
[001395] Step 2: 1 -(4-Chloro-3-(3-
Figure imgf000287_0001
[001396] To a solution of methyl 1 -(4-chloro-3-(3- methoxypropoxy)benzyl)cyclopentane-1 -carboxylate (2.28 g, 6.69 mmol) in THF :MeOH (10 mL : 20 mL) was added LiOH (1 .28 g, 53.5 mmol) in water (10 mL) and warmed to 55 °C. After 3 hrs. the mixture was poured into 100 mL HCI (1 N aq) and extracted with EtOAc. The organic phase was dried (brine, Na2S04) and concentrated to give 1 -(4-chloro-3-(3- methoxypropoxy)benzyl)cyclopentane-1 -carboxylic acid (2.06 g, 6.30 mmol, 94 % yield) an orange oil. LCMS (ESI) m/z: 349.4 (M + 1)+.
[001397] Step 3: N-(1 -(4-Chloro-3-(3- methoxypropoxy)benzyl)cvclopentyl)formamide
Figure imgf000287_0002
[001398] To 1 -(4-chloro-3-(3-methoxypropoxy)benzyl)cyclopentane-1 - carboxylic acid (1 .14 g, 3.49 mmol) was added TEA (0.583 mL, 4.19 mmol), tert-butanol (10 mL) and DPPA (0.902 mL, 4.19 mmol). The reaction was heated to 80 °C under nitrogen for 1 hr and poured into 1 N HCI (aq) at 80 °C. A precipitate formed. Dioxane (10 mL) was added stirred for 30 minutes, cooled, poured into NaHC03 (sat aq), and extracted with EtOAc. The organic phase was dried (brine, Na2S04) and concentrated. The residue was then dissolved in1 ,4-dioxane (40 mL) before a pre-mixed solution of formic acetic mixed anhydride (6.8 M in formic acid) 2 mL was added. The suspension was heated until all solids dissolved and then cooled to RT. Another 5 mL of anhydride was added and the mixture heated to 80 °C for one hour. The mixture was concentrated to give a yellow oil that was used directly in the next step. LCMS (ESI) m/z: 325.4 (M -1)".
[001399] Step 4: 7'-Chloro-6'-(3-methoxypropoxy)-4'H-spiro[cvclopentane-1 ,3'- isoquinolinel
Figure imgf000287_0003
[001400] To N-(1 -(4-chloro-3-(3- methoxypropoxy)benzyl)cyclopentyl)formamide (2.26 g, 6.94 mmol) in acetonitrile (50 mL) was added POCI3 (0.646 mL, 6.94 mmol) and the mixture heated to 80 °C. After 30 minutes the reaction was added to ice and partitioned between NaHC03 (aq) and EtOAc. The aqueous phase was removed and washed again with EtOAc. The organics were combined, dried (brine, Na2S04), and concentrated to give 7'-chloro-6'-(3-methoxypropoxy)-4'H- spiro[cyclopentane-1 ,3'-isoquinoline] (2.135 g, 6.94 mmol, 1 00 % yield) as a red oil. LCMS (ESI) m/z: 308.3 (M + 1 )+.
[001401] Step 5: Ethyl 10l-chloro-9'-(3-methoxypropoxy)-2l-oxo-1 '.Ι'.ΤΛ 1 b1- tetrahydrospirofcyclopentane-1 ,6'-pyrido[2, 1 -alisoquinolinel-3'-carboxylate
Figure imgf000288_0001
[001402] A solution of 7'-chloro-6'-(3-methoxypropoxy)-4'H- spiro[cyclopentane-1 ,3'-isoquinoline] (2.14 g , 6.94 mmol) and ethyl 2-(ethoxymethylene)-3- oxobutanoate (4.83 ml, 27.7 mmol) in ethanol was heated to 80 °C for 16 hr. The mixture was cooled, partitioned between water and EtOAc, and extracted with additional EtOAc. The organic phase was dried (brine, Na2S04) and concentrated to give ethyl 1 0'-chloro-9'- (3-methoxypropoxy)-2'-oxo-1 ',2',7', 1 1 b'-tetrahydrospiro[cyclopentane-1 ,6'-pyrido[2, 1 - a]isoquinoline]-3'-carboxylat as a red oil. LCMS (ESI) m/z: 448.3. (M + 1 )+.
[001403] Step 6: Ethyl 10'-chloro-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihydrospirofcyclopentane-1 ,6'-pyrido[2, 1 -alisoquinolinel-3'-carboxylate
Figure imgf000288_0002
[001404] A solution of ethyl 10'-chloro-9'-(3-methoxypropoxy)-2'-oxo- 1 ',2',7', 1 1 b'-tetrahydrospiro[cyclopentane-1 ,6'-pyrido[2, 1 -a]isoquinoline]-3'-carboxylate (3.1 1 g, 6.94 mmol) and DDQ (3.94 g, 17.36 mmol) in 1 ,2-dimethoxyethane (DME) (40 mL) was heated to 80 °C for 3hr. The mixture was partitioned between NaHC03 (Aq) and EtOAc. The organics were concentrated and the residue purified on silica (0-1 0% MeOH in DCM) twice to give ethyl 10'-chloro-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclopentane-1 ,6'- pyrido[2, 1 -a]isoquinoline]-3'-carboxylate (230 mg, 0.516 mmol, 7.43 % yield) as a dark brown solid. LCMS (ESI) m/z: 446.3. (M + 1 )+. [001405] Step 7: 10'-Chloro-9'-(3-methoxypropoxy)-3.3-dimethyl-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000289_0001
[001406] A solution of ethyl 10'-chloro-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospiro[cyclopentane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (120 mg, 0.269 mmol) in methanol (10 mL) andl ,4-dioxane (5 mL) was treated with LiOH (77 mg, 3.23 mmol) in water (5.00 mL) and heated to 60 °C for 1 hr. The solution was concentrated and partitioned between HCI (1 N) and EtOAc. The organics were dried (Na2S04, brine) and purified by reverse phase HPLC to give 10'-chloro-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospiro[cyclopentane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (35.4 mg, 0.085 mmol, 31 .5 % yield) as a light tan solid. 1 H NMR (400 MHz, c/6-DMSO-d6) δ ppm 8.50 (s, 1 H), 8.18 (s, 1 H), 7.42 (s, 1 H), 7.20 (s, 1 H), 4.19 (t, J= 6.2 Hz 2 H), 3.49 (t, J = 6.2 Hz, 2H), 3.23 (s, 3H), 3.22 (s, 2 H), 2.12 - 1 .67 (m, 10 H). LCMS(ES+) (m/z):418.6 (M + 1 )+.
[001407] Example 123
[001408] 10'-chloro-9'-(cyclopropylmethoxy)-1 '-fluoro-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000289_0002
[001409] Step 1 : 3-(Benzyloxy)-4-chlorobenzaldehyde
[001410] To a solution of 4-chloro-3-hydroxybenzaldehyde (15 g, 96 mmol) in Ν,Ν-dimethylformamide (DMF) (200 mL) was added potassium carbonate (66.2 g, 479 mmol) and (bromomethyl)benzene (13.7 mL, 1 15 mmol). The reaction was stirred at room temp overnight. The suspension was filtered, washed with Et20 (2 x 50 mL) and the combined solution washed with water (100 mL). The aqueous layer was washed with Et20 (2x 50 mL). The combined organics were washed with LiCI (aq 5%, 50 mL), dried with brine and Na2S04, and concentrated to give an off-white solid. The solid was recrystallized from 300 mL hexanes to give 3-(benzyloxy)-4-chlorobenzaldehyde (18.9 g, 77 mmol, 80 % yield) as slightly pink crystals. LCMS(ES+) (m/z):247.2 (M + 1)+.
[001411] Step 2: (3-(Benzyloxy)-4-chlorophenyl)methanol
Figure imgf000290_0001
[001412] To a solution of 3-(benzyloxy)-4-chlorobenzaldehyde (18.7 g, 76 mmol) in methanol (200 mL) at 0 °C was added sodium borohydride (5.74 g, 152 mmol) portion wise and the mixture allowed to warm to RT. The mixture was stirred for 1 hr, poured into half sat NH4CI (aq. 150 mL), and filtered. The filtration was slow, so the remaining slurry (~1/2 filtered) was treated with Et20 (200 mL) for two solution phases. The organics were dried (brine, Na2S04) and concentrated to give (3-(benzyloxy)-4- chlorophenyl)methanol (19.47 g, 78 mmol, 103 % yield) as a white solid. LCMS(ES+) (m/z):247.2 (M-H20 + H+).
[001413] Step 3: 2-(Benzyloxy)-4-(bromomethyl)-1 -chlorobenzene
Figure imgf000290_0002
[001414] To a solution of (3-(benzyloxy)-4-chlorophenyl)methanol (5.73 g,
23.0 mmol) in tetrahydrofuran (THF) (100 mL) at RT was added TMS-Br (4.48 mL, 34.6 mmol) and the mixture stirred for 4 hours. An additional aliquot of TMS-Br (4.48 mL, 34.6 mmol) was added and the mixture stirred at RT for 18h. Phosphorus tribromide (3.26 mL, 34.6 mmol) was added dropwise and the mixture stirred at ambient temperature. The mixture grew warm during addition. After 5 min. the reaction was cooled to 0 °C, slowly poured onto a mixture of ice and sat NaHCC>3 (aq), and the extracted with Et20. The combined organics were dried (brine, Na2S04), evaporated onto celite, and purified by silica chromatography (0-25% EtOAc in hexanes) to give 2-(benzyloxy)-4-(bromomethyl)-1 - chlorobenzene (5 g, 16.05 mmol, 69.6 % yield) as a white solid. 1H NMR (400 MHz, CDCI3) δ 7.50-7.30(m, 6H), 7.01 (s, 1 H), 6.95 (m, 1 H), 5.18 (s, 2H), 4.45 (s, 2H).
Figure imgf000290_0003
[001416] To LDA (46.5 ml, 46.5 mmol) at 0 °C was added ethyl
cyclobutanecarboxylate (4.77 g, 37.2 mmol) and the mixture stirred for 5 min. A solution of 2-(benzyloxy)-4-(bromomethyl)-1 -chlorobenzene (5.8 g, 18.61 mmol) in THF (50mL) was added dropwise and the mixture allowed to warm to RT. The mixture was quenched with ice and NH4CI (aq) and extracted with EtOAc. The organics were combined, dried (brine, Na2S04), and concentrated to an oil. The crude oil was purified on silica gel hex:EtOAc (eluting at ~10%) to give ethyl 1 -(3-(benzyloxy)-4-chlorobenzyl)cyclobutane-1 -carboxylate (3.7 g, 10.31 mmol, 55.4 % yield) as a clear oil with some white crystals. LCMS(ES+) (m/z): 357.8 (M - 1)-.
[001417] hlorobenzyl)cvclobutanecarboxylic acid
Figure imgf000291_0001
[001418] To ethyl 1 -(3-(benzyloxy)-4-chlorobenzyl)cyclobutane-1 -carboxylate (3.7 g, 10.31 mmol) in methanol : water 1 :1 50 mL was added lithium hydroxide (0.247 g, 10.31 mmol) and the mixture stirred at 60 °C for 16h. The reaction was cooled, acidified with HCI (1 N) and extracted with EtOAc. The organics were combined, dried (brine, Na2S04), concentrated, and purified on silica (0-60 % EtOAc:Hex) to yield 1 -(3-(benzyloxy)-4- chlorobenzyl)cyclobutane-1 -carboxylic acid (2.1 g, 6.35 mmol, 61 .6 % yield) as a clear oil that solidified upon standing. LCMS (ESI) m/z: 329.3 (M-1)-.
[001419] -chlorobenzyl)cvclobutyl)formamide
Figure imgf000291_0002
[001420] To a solution of 1 -(3-(benzyloxy)-4-chlorobenzyl)cyclobutane-1 - carboxylic acid (2.1 g, 6.35 mmol) in toluene (50 mL) was added TEA (1 .062 mL, 7.62 mmol) followed by diphenyl phosphorazidate (2.10 g, 7.62 mmol). The mixture was stirred at 90 °C for 2 h. After the reaction was finished the mixture was cooled to room temperature and quenched with concentrated HCI (5 mL). The mixture was heated to 60°C for 2 h. The mixture was cooled to 0 °C and basified with 1 N NaOH aqueous. The organic layer was washed with brine and dried over MgS04. To the solution of 1 -(3-(benzyloxy)-4- chlorobenzyl)cyclobutan-1 -amine (1 .91 g, 6.33 mmol) in toluene (50 mL) was added formic acetic anhydride (from 18mL Ac20 + 10 mL formic acid) (2 mL) and stirred at RT for 45 min. The solvents were evaporated to give crude N-(1 -(3-(benzyloxy)-4- chlorobenzyl)cyclobutyl)formamide (2.09 g, 6.33 mmol, 100 % yield). LCMS (ESI) m/z: 330.3 (M + 1)+.
[001421] Step 7: 6'-(Benzyloxy)-7'-chloro-4'H-spiro[cyclobutane-1 ,3'- isoquinolinel
Figure imgf000292_0001
[001422] To a solution of N-(1 -(3-(benzyloxy)-4- chlorobenzyl)cyclobutyl)formamide (2.09 g, 6.33 mmol) was added POCI3 (0.705 mL, 7.56 mmol) dropwise and the mixture stirred at RT for 60 minutes. The mixture was basified with ammonium hydroxide to pH = 9 and extracted with DCM. The organic phase was dried (brine, Na2S04) and concentrated to give crude 6'-(benzyloxy)-7'-chloro-4'H- spiro[cyclobutane-1 ,3'-isoquinoline] (1 .964 g, 6.30 mmol, 100 % yield). LCMS (ESI) m/z: 312.3 (M + 1)+.
[001423] Step 8: Ethyl 9'-(benzyloxy)-10'-chloro-l '-fluoro^'-oxo^'J'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000292_0002
[001424] A mixture of zinc iodide (989 mg, 3.10 mmol) powder and 6'- (benzyloxy)-7'-chloro-4'H-spiro[cyclobutane-1 ,3'-isoquinoline] (967 mg, 3.1 mmol) in MeCN (30 mL) was treated with a crude solution of ethyl (Z)-2-(ethoxymethylene)-4,4-difluoro-3- ((trimethylsilyl)oxy)but-3-enoate (5475 mg, 18.60 mmol) in DMF (30 mL) and the reaction heated at 50 °C overnight. The reaction was cooled, treated with HCI (10% aq), and extracted with DCM. The organic layers were combined, dried (brine, Na2S04), filtered, and concentrated. The crude was purified on silica eluting with 0-10% MeOH:EtOAc to give ethyl 9'-(benzyloxy)-10'-chloro-1 '-fluoro-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (280 mg, 0.598 mmol, 19.30 % yield). LCMS (ESI) m/z: 468.3 (M + 1 ).
[001425] Step 1 1 : Ethyl 10'-chloro-9'-(cvclopropylmethoxy)-1 '-fluoro-2'-oxo- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000292_0003
[001426] A solution of ethyl 9'-(benzyloxy)-10'-chloro-1 '-fluoro-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (280 mg, 0.598 mmol) in trifluoroacetic acid (TFA) (100 ml_) was heated to 65 °C for 6 h. The solvents were evaporated and the residue dissolved in Ν,Ν-dimethylformamide (DMF) (5 ml_). The mixture was treated with cesium carbonate (293 mg, 0.900 mmol) and (bromomethyl)cyclopropane (97 mg, 0.720 mmol) before being heated to 65 °C for 30 min. The mixture was stirred at RT overnight. Another 100 mg (bromomethyl)cyclopropane was added and the mixture heated to 75 °C for 3h. The mixture was partitioned between EtOAc and LiCI (aq), the aq. layer was reextracted with EtOAc, and the organics combined. The organic phase was washed with LiCI (aq), dried (Na2S04), concentrated, and purified on silica (EtOAc:0-10% MeOH) to give 38 mg ethyl 10'-chloro-9'-(cyclopropylmethoxy)-1 '-fluoro-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate as a ~1 :1 mixture of SM and desired product. LCMS (ESI) m/z: 432.3 (M + 1)+.
[001427] Step 12: 10'-Chloro-9'-(cvclopropylmethoxyV1 '-fluoro-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000293_0001
[001428] A solution of ethyl 10'-chloro-9'-(cyclopropylmethoxy)-1 '-fluoro-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (33 mg, 0.076 mmol) and lithium hydroxide (9.15 mg, 0.382 mmol) in water : MeOH : MeCN (ca 1 :1 :1 , 5 ml) was heated to 62 °C for 1 hr. The reaction was cooled, acidified (TFA), and purified on reverse phase HPLC to give10'-chloro-9'-(cyclopropylmethoxy)-1 '-fluoro-2'-oxo- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (3.5 mg, 8.67 Mmol, 1 1 .34 % yield). Ή NMR (400 MHz, CDCI3) δ ppm 8.84 (s, 1 H), 8.1 1 (s, 1 H), 6.85 (s, 1 H), 3.99 (d, J=8 Hz, 2H), 3.21 (s, 2 H), 2.62 (m,2H), 2.03 (s, 4H), 1 .37 (m, 1 H), 0.72 (m, 2H), 0.44 (m, 2H). LCMS (ESI) m/z: 404.4 (M + 1)+.
[001429] Example 124
[001430] 10'-chloro-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000293_0002
Stepl : Ethyl 1 -(4-chloro-3-(3-methoxypropoxy)benzyl)cvclobutanecarboxylate
Figure imgf000294_0001
[001431] A solution of n-butyllithium (7.19 ml, 17.98 mmol) was added dropwise to a -78 °C solution of diisopropylamine (2.56 ml, 17.98 mmol) in THF (20.0 ml). The mixture was warmed to room temperature, and was stirred for 15 minutes before being cooled down to -78 °C. Ethyl cyclobutanecarboxylate (2.305 g, 17.98 mmol) was added dropwise (neat), and the mixture was stirred at 0°C for 30 minutes. The reaction mixture was then cooled to -78 °C before dropwise addition of 4-(bromomethyl)-1 -chloro-2-(3- methoxypropoxy)benzene (4.4g, 14.99 mmol) in solution of THF (10.00 ml). The reaction mixture was stirred for 40 minutes at -78 °C before being warmed to 0 °C and then quenched with water. EtOAc was added and the organic layer was separated and dried over MgS04. The solvent was evaporated to give 5.4 grams of crude ethyl 1 -(4-chloro-3-(3- methoxypropoxy)benzyl)cyclobutanecarboxylate as an oil which was directly used in the next step without purification. LCMS (ESI) m/z: 341 .3(M+1 )+.
[001432] Step 2: 1 -(4-Chloro-3-(3-methoxypropoxy)benzyl)cvclobutane-1 - carboxylic acid
Figure imgf000294_0002
[001433] The crude ethyl 1 -(4-chloro-3-(3- methoxypropoxy)benzyl)cyclobutanecarboxylate (5.4 g) from previous step was dissolved in THF (40.00 ml), methanol (20.00 ml), water (20.00 ml) and was treated with LiOH (5.03 g, 120 mmol) at 55 °C for 5 hours. The reaction mixture was acidified with 1 N HCI to pH 4, and the mixture was concentrated. Water and EtOAc were added and the organic layer was separated. The solvent was evaporated and the crude material was purified via silica gel chromatography (5-30 % EtOAc/Hex) to give 1 -(4-chloro-3-(3- methoxypropoxy)benzyl)cyclobutane-1 -carboxylic acid (3.19 g, 10.20 mmol, 68.0 % yield). LCMS (ESI) m/z: 313.2 (M+1)+.
[001434] Step 3: 1 -(4-Chloro-3-(3-methoxypropoxy)benzyl)cyclobutan-1 - amine
Figure imgf000294_0003
[001435] A solution of 1 -(4-chloro-3-(3-methoxypropoxy)benzyl)cyclobutane-
1 -carboxylic acid (2.66 g, 8.50 mmol), DPPA (2.199 ml, 10.20 mmol) and Et3N (1 .422 ml, 10.20 mmol) in toluene (20 ml) was stirred for 1 hour at 80 °C. The mixture was cooled to room temperature, 1 ,4-dioxane (20.00 ml) and HCI (6M) (14.17 ml, 85 mmol) were added and the reaction mixture was stirred at 80 °C for 90 minutes. The mixture was cooled to 0 °C and was basified with NaOH to pH 12. EtOAc and water were added, the organic layer was separated, dried over MgS04, and was concentrated to give 2.5 g crude 1 -(4-chloro-3-(3- methoxypropoxy)benzyl)cyclobutan-1 -amine. LCMS (ESI) m/z: 284.2 (M+1 )+.
[001436] Step 4: N-(1 -(4-chloro-3-(3- methoxypropoxy)benzyl)cvclobutyl)formamide
Figure imgf000295_0001
[001437] Crude 1 -(4-chloro-3-(3-methoxypropoxy)benzyl)cyclobutan-1 -amine 2.5 g was dissolved in DCM (20.00 ml), cooled to 0 °C and was treated with Et3N (1 .4 ml, 10.20 mmol) and acetic formic anhydride (6.35 ml, 42.5 mmol) for 1 hour. The solvent was evaporated and the crude material was purified via silica gel chromatography to give N-(1 - (4-chloro-3-(3-methoxypropoxy)benzyl)cyclobutyl)formamide (2.35 g, 7.54 mmol, 89 % yield). LCMS (ESI) m/z: 312.2 (M+1)+.
[001438] Step 5: 7'-chloro-6'-(3-methoxypropoxy)-4'H-spiro[cvclobutane-1 ,3'- isoquinolinel
Figure imgf000295_0002
[001439] N-(1 -(4-chloro-3-(3-methoxypropoxy)benzyl)cyclobutyl)formamide (1 .99 g, 6.39 mmol) was dissolved in acetonitrile (20 mL) and was heated with POCI3 (0.59 mL, 6.39 mmol) at 90 °C for one hour. The reaction mixture was concentrated, diluted with EtOAc, cooled to 0 °C and was basified with NH4OH until pH > 1 1 . The organics were separated, dried over MgS04 and concentrated to give 2.1 g crude 7'-chloro-6'-(3- methoxypropoxy)-4'H-spiro[cyclobutane-1 ,3'-isoquinoline] which was used in the next reaction without purification. LCMS (ESI) m/z: 294.2 (M+1)+.
[001440] Step 6: ethyl 10'-chloro-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000295_0003
[001441] Crude 7'-chloro-6'-(3-methoxypropoxy)-4'H-spiro[cyclobutane-1 ,3'- isoquinoline] (2.1 g) was dissolved in ethanol (10.00 mL) and was treated with ethyl (Z)-2- (ethoxymethylene)-3-oxobutanoate (4.76 g, 25.5 mmol) with heating at 84 °C overnight. The reaction mixture was concentrated, dissolved in 1 ,2-Dimethoxyethane (DME) (20.00 mL) and the mixture was treated with 2,3,5,6-tetrachlorocyclohexa-2,5-diene-1 ,4-dione (1 .57 g, 6.39 mmol) at 90 °C for 1 hour. The reaction mixture was concentrated, and the crude material was purified via silica gel chromatography to give ethyl 10'-chloro-9'-(3- methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (1 .4 g, 3.24 mmol, 50.8 % yield). LCMS (ESI) m/z: 432.3 (M+1 )+.
[001442] Step 7: 10'-chloro-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihydrospirofcyclobuta -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000296_0001
[001443] A solution of ethyl 10'-chloro-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (106 mg, 0.245 mmol) and LiOH (1 .0M solution) (0.98 mL, 0.98 mmol) in MeOH (10.00 mL) at rt overnight, The reaction mixture was quenched with 1 N HCI, concentrated, and was diluted with EtOAc and water. The organics were separated, washed with brine, dried over MgS04 and concentrated. The residue was suspended in DME and the solid was filtered off to give 86 mg of crude material which was suspended in 2.0 mL EtOAc and filtered. The solid was washed with an additional 2.0 mL cold EtOAc to give 10'-chloro-9'-(3-methoxypropoxy)-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (56 mg, 0.14 mmol, 57 % yield). LCMS (ESI) m/z: 404.3 (M+1 )+. 1 H NMR (400 MHz,
CHLOROFORM-d) d ppm 8.93 (s, 1 H), 7.73 (s, 1 H), 7.04 (s, 1 H), 6.88 (s, 1 H), 4.22 (t, J = 6.1 Hz, 2 H), 3.62 (t, J=5.7 Hz, 2 H), 3.37 (s, 3 H), 3.26 (s, 2 H), 2.57 - 2.71 (m, 2 H), 1 .91 - 2.23 (m, 6 H).
[001444] Example 125
[001445] 9'-(3-hvdroxypropoxy)-10'-methoxy-2'-oxo-2',7'- dihydrospirofcyclobutan -1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000296_0002
[001446] A solution of ethyl 9'-hydroxy-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (75 mg, 0.21 mmol), 3-bromopropan-1 -ol (88 mg, 0.63 mmol) and K2C03 (87 mg, 0.63 mmol) in DME (5 mL) was stirred at 60 °C for 5 hours. EtOAc and water were added, the organics were separated, concentrated. This residue was dissolved in ethanol (5.00 mL) and was treated with LiOH (1 .0 M solution) (1 .06 mL, 1 .06 mmol) at rt for 6 hours.The reaction was quenched with 1 M HCI to pH = 3, and the mixture was concentrated. The residue was dissolved in EtOAc, water was added, the organic layer was separated and dried over MgS04 and concentrated. The crude material was purified via reverse phase HPLC to give 9'-(3-hydroxypropoxy)-10'- methoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (21 mg, 0.052 mmol, 24 % yield). LCMS (ESI) m/z: 386.3 (M+1)+. 1 H NMR (400 MHz, CHLOROFORM-d) d ppm 8.92 (s, 1 H), 7.72 (s, 1 H), 7.03 (s, 1 H), 6.87 (s, 1 H), 4.21 (t, J = 6.2 Hz, 2 H), 3.60 (t, J = 5.9 Hz, 2 H), 3.36 (s, 3 H), 3.22 (s, 2 H), 2.56 - 2.70 (m, 2 H), 1 .92 - 2.21 (m, 6 H).
[001447] Example 126
[001448] 10'-chloro-9'-(cvclopropylmethoxyV2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000297_0001
[001449] Stepl : Ethyl 10'-chloro-g'-hvdroxy^'-oxo^'J1- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate and 10'-chloro-9'- hvdroxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000297_0002
[001450] To a solution of ethyl 10'-chloro-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (733 mg, 1 .70 mmol) in DCM (15 mL) was added BBr3 (0.80 mL, 8.49 mmol) and the mixture was stirred at room temperature for 3 hours. The reaction mixture was filtered to give 883 mg of a tan solid as a mixture of co-eluting ethyl 10'-chloro-9'-hydroxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylate (883 mg, 2.45 mmol, 145 % yield) and 10'-chloro- 9'-hydroxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid which were used in the next step as is. LCMS (ESI) m/z: 332.2 (M+1)+ for 10'-chloro-9'- hydroxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid and LCMS (ESI) m/z: 360.2 (M+1 )+ for ethyl 10'-chloro-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate.
[001451] Step 2: 10'-chloro-9'-(cvclopropylmethoxy)-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000298_0001
[001452] A mixture of 90 mg ethyl 10'-chloro-9'-hydroxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate and 10'-chloro-9'- hydroxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid in DMF (1 .00 mL) was treated with (bromomethyl)cyclopropane (169 mg, 1 .25 mmol) and K2C03 (173 mg, 1 .25 mmol) at 60 °C for 5 hours. Water and EtOAc were added, and the organics were separated, dried over MgS04 and was concentrated. The crude material was dissolved in methanol (3.00 mL) and was treated with LiOH (2.50 mL, 2.50 mmol) at 40 °C for 4 hours. The reaction mixture was acidified with aqueous HCI, EtOAc was added, and the organics were separated. The organics were concentrated and the crude material was purified via reverse phase HPLC to give 10'-chloro-9'-(cyclopropylmethoxy)-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (25 mg, 0.062 mmol, 25 % yield). LCMS (ESI) m/z: 386.2 (M+1)+. 1 H NMR (400 MHz, CHLOROFORM-d) d ppm 8.88 (s, 1 H), 7.72 (s, 1 H), 7.00 (s, 1 H), 6.81 (s, 1 H), 3.96 (d, J = 6.6 Hz, 2 H), 3.21 (s, 2 H), 2.54 - 2.70 (m, 2 H), 2.09 - 2.19 (m, 2 H), 1 .90 - 2.09 (m, 2 H), 1 .27 - 1 .42 (m, 1 H), 0.63 - 0.76 (m, 2 H), 0.33 - 0.47 (m, 2 H)
[001453] Examples'! 27
[001454] 9-(Cvclopropylmethoxy)-10-(difluoromethyl)-1 -fluoro-6-isopropyl-6- methyl-2-oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid (isomer 1 )
[001455] Example 128
[001456] 9-(Cvclopropylmethoxy)-10-(difluoromethyl)-1 -fluoro-6-isopropyl-6- methyl-2-oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid (isomer 2)
Figure imgf000299_0001
Example 127 Example 128
[001457] Step 1 : Ethyl (ZV2-(ethoxymethyleneV4.4-difluoro-3- ((trimethylsilyl)oxy)butanoate
Figure imgf000299_0002
[001458] A suspension of TMSCI (19.43 ml_, 152 mmol) and magnesium (3.69 g, 152 mmol) was sonicated for 15 minutes. DMF (30 ml_) and ethyl (Z)-2- (ethoxymethylene)-4,4,4-trifluoro-3-oxobutanoate (4.56 g, 19 mmol) were added dropwise at 50 °C and was stirred for another 3 minutes at 50 °C. Excess TMSCI was removed under reduced pressure and the mixture was filtered. The filter cake was washed with 20 ml_ of DMF. This resulted in a 19 mmol / 50 ml_ = 0.38 M solution in DMF which was used in the next step as is.
[001459] Step 2: Ethyl 9-(benzyloxy)-10-bromo-1 -fluoro-6-isopropyl-6-methyl- 2-oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000299_0003
[001460] A suspension of 6-(benzyloxy)-7-bromo-3-isopropyl-3-methyl-3,4- dihydroisoquinoline (403 mg, 1 .08 mmol) and zinc iodide (346 mg, 1 .08 mmol) in MeCN (3ml_) was placed in a 50 °C oil bath. Ethyl (Z)-2-(ethoxymethylene)-4,4-difluoro-3- ((trimethylsilyl)oxy)butanoate (0.38 M solution in DMF) (1 1 .39 ml_, 4.33 mmol) was added dropwise and the reaction was stirred overnight at 50 °C. The reaction mixture was cooled to room temperature, diluted with EtOAc and was quenched with 1 N HCI. This resulted in a biphasic mixture with suspended solid particles. The organic layer was separated, and contained small amounts of desired product as judged by LCMS. The water layer containing solid particles was filtered and ethyl 9-(benzyloxy)-10-bromo-1 -fluoro-6-isopropyl-6-methyl- 2-0X0-6, 7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (315 mg) was collected as a cream solid. LCMS (ESI) m/z: 528.2/530.2 (M/M+2)+.
[001461] Step 3: Ethyl 10-bromo-1 -fluoro-9-hvdroxy-6-isopropyl-6-methyl-2- oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000300_0001
[001462] A solution of ethyl 9-(benzyloxy)-10-bromo-1 -fluoro-6-isopropyl-6- methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (315 mg) in TFA (5 ml_, 64.9 mmol) was stirred at 70 °C for 2 hours. The reaction mixture was cooled to room temperature and concentrated. Water was added and 261 mg ethyl 10-bromo-1 -fluoro-9- hydroxy-6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate was collected by filtration as an off-white solid, and was used as is in the next step without purification. LCMS (ESI) m/z: 438.2/440.2 (M/M+2)+.
[001463] Step 4: Ethyl 10-bromo-9-(cvclopropylmethoxy)-1 -fluoro-6-isopropyl-
Figure imgf000300_0002
[001464] Ethyl 10-bromo-1 -fluoro-9-hydroxy-6-isopropyl-6-methyl-2-oxo-6,7- dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (261 mg), (bromomethyl)cyclopropane (322 mg, 2.38 mmol) and K2C03 (247 mg, 1 .79 mmol) in DMF (4 mL) was stirred at 80 °C for 1 .5 hours. The reaction mixture was cooled to room temperature, water was added and 264 mg ethyl 10-bromo-9-(cyclopropylmethoxy)-1 -fluoro-6-isopropyl-6-methyl-2-oxo-6,7- dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate was collected by filtration as a cream solid. LCMS (ESI) m/z: 492.2/494.2 (M/M+2)+.
[001465] Step 5: Ethyl 9-(cyclopropylmethoxy)-1 -fluoro-6-isopropyl-6-methyl- 2-OXO-10-vinyl-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000301_0001
[001466] A solution of crude ethyl 10-bromo-9-(cyclopropylmethoxy)-1 -fluoro- 6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (260 mg), PdCI2(dppf)-CH2CI2 adduct (43.1 mg, 0.053 mmol), triethylamine (0.368 ml_, 2.64 mmol) and potassium vinyl trifluoroborate (141 mg, 1 .06 mmol) in EtOH (5 ml_) was stirred for 2 hours at 95 °C. The reaction mixture was concentrated, water was added, and 286 mg of a dark solid was filtered off as ethyl 9-(cyclopropylmethoxy)-1 -fluoro-6-isopropyl-6-methyl- 2-OXO-10-vinyl-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate. This was used in the next step without purification. LCMS (ESI) m/z: 440.3 (M+1)+.
[001467] Step 6: Ethyl 9-(cyclopropylmethoxy)-1 -fluoro-1 O-formyl-6-isopropyl- 6-methyl-2-oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000301_0002
[001468] To a suspension of ethyl 9-(cyclopropylmethoxy)-1 -fluoro-6- isopropyl-6-methyl-2-oxo-10-vinyl-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (240 mg) in THF (20 ml_) and water (0.2 ml_) was added potassium osmate dihydrate (240 mg, 0.65 mmol) and sodium periodate (934 mg, 4.37 mmol) and the reaction mixture was stirred at room temperature. The solvent was evaporated, water was added and 260 mg ethyl 9-(cyclopropylmethoxy)-1 -fluoro-10-formyl-6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H- pyrido[2,1 -a]isoquinoline-3-carboxylate containing osmium salts were filtered off as an ashy solid LCMS (ESI) m/z: 442.4 (M+1 )+.
[001469] Step 7: Ethyl 9-(cyclopropylmethoxy)-10-(difluoromethyl)-1 -fluoro-6- isopropyl-6-methyl-2-o -6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000301_0003
[001470] A solution of crude ethyl 9-(cyclopropylmethoxy)-1 -fluoro-10-formyl- 6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (220 mg) and DAST (0.13 ml_, 0.99 mmol) in DCM (10 mL) was stirred overnight at room
temperature. More DAST (0.13 mL, 0.99 mmol) was added and the reaction was stirred for another 6 hours. The reaction mixture was diluted with saturated sodium bicarbonate and the organics were separated, dried over MgS04 and concentrated to give 234 mg of crude material which was used in the next step without purification. LCMS (ESI) m/z: 464.4 (M+1)+.
[001471] Step 8: 9-(Cvclopropylmethoxy)-10-(difluoromethyl)-1 -fluoro-6- isopropyl-6-methyl-2-o -6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000302_0001
[001472] The crude (ethyl 9-(cyclopropylmethoxy)-10-(difluoromethyl)-1 - fluoro-6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-py rido[2,1 -a]isoquinoline-3-carboxylate (234 mg) was dissolved in THF (5 mL), Water (3 mL), Ethanol (5.00 mL) and was treated with lithium hydroxide (85 mg, 2.02 mmol) overnight. The reaction mixture was acidified with 1 N HCI and the mixture was concentrated. Water and EtOAc were added, the organic layer was separated, concentrated and was purified via reverse phase HPLC (10-90 % gradient MeCN, H20, 0.1 % FA modifier) to give 9-(cyclopropylmethoxy)-10-(difluoromethyl)-1 -fluoro- 6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylic acid (23 mg, 0.053 mmol, 1 1 % yield). LCMS (ESI) m/z: 436.3 (M+1)+. 1 H NMR (400 MHz,
CHLOROFORM-d) d ppm 8.58 (s, 1 H), 8.33 (s, 1 H), 6.82 - 7.16 (m, 1 H), 6.77 (s, 1 H), 3.95 (d, J = 7.0 Hz, 2 H), 3.01 - 3.24 (m, 2 H), 1 .75 - 1 .90 (m, 1 H), 1 .68 (s, 3 H), 1 .18 - 1 .36 (m, 1 H), 0.83 (d, J = 7.0 Hz, 3 H), 0.64 - 0.74 (m, 5 H), 0.38 (q, J=4.9 Hz, 2 H).
[001473] Step 9: 9-(cvclopropylmethoxy)-10-(difluoromethyl)-1 -fluoro-6- isopropyl-6-methyl-2-oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
[001474] Racemic 9-(cyclopropylmethoxy)-10-(difluoromethyl)-1 -fluoro-
6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylic acid was separated via chiral chromatography to give 9-(cyclopropylmethoxy)-10-(difluoromethyl)-1 - fluoro-6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylic acid (Example 127, isomer 1 ) and 9-(cyclopropylmethoxy)-10-(difluoromethyl)-1 -fluoro-6- isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylic acid (Example 128, isomer 2).
Figure imgf000303_0001
Example 127 Example 128
[001475] 9-(cyclopropylmethoxy)-10-(difluoromethyl)-1-fluoro-6-isopropyl-6- methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1-a]isoquinoline-3-carboxylic acid (Example 127, isomer 1) 1 H NMR (400 MHz, CHLOROFORM-d) d ppm 8.59 (s, 1 H), 8.33 (s, 1 H), 6.82 - 7.16 (m, 1 H), 6.78 (s, 1 H), 3.95 (d, J = 6.6 Hz, 1 H), 3.05 - 3.27 (m, 1 H), 1 .76 - 1.92 (m, 1 H), 1 .69 (s, 1 H), 1 .18 - 1.35 (m, 1 H), 0.77 - 0.90 (m, 1 H), 0.61 - 0.75 (m, 2 H), 0.38 (q, J=5.1 Hz, 1 H). LCMS (ESI) m/z: 436.3 (M+1)+. 9-(cyclopropylmethoxy)-10-(difluoromethyl)- 1-fluoro-6-isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylic acid (Example 128, isomer 2) 1 H NMR (400 MHz, CHLOROFORM-d) d ppm 8.59 (s, 1 H), 8.33 (s, 1 H), 6.82 - 7.15 (m, 1 H), 6.78 (s, 1 H), 3.95 (d, J = 6.6 Hz, 2 H), 3.04 - 3.24 (m, 2 H), 1 .77 - 1 .92 (m, 1 H), 1 .69 (s, 3 H), 1.18 - 1.33 (m, 1 H), 0.83 (d, J = 6.6 Hz, 3 H), 0.62 - 0.76 (m, 5 H), 0.38 (q, J=4.9 Hz, 2 H). LCMS (ESI) m/z: 436.3 (M+1)+.
[001476] Example 129
[001477] 9'-(CvclopropylmethoxyV10'-(difluoromethylV1 '-fluoro-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1-alisoquinolinel-3'-carboxylic acid
Figure imgf000303_0002
[001478] Step 1 : Ethyl 1 '-fluoro-10'-formyl-g'-hvdroxy^'-oxo^'J1- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000303_0003
[001479] Ethyl 9'-(benzyloxy)-1 '-fluoro-10'-formyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (59 mg, 0.128 mmol) in TFA (1 mL) was mixed and heated at 80°C for 3 hours. The mixture was concentrated under reduced pressure, diluted with ice water and ethyl acetate. The mixture was extraced with ethyl acetate, the combined organics were dried over sodium sulfate and concentrated under reduced pressure to give a brown solid (94 mg) that was used in the next step without further purification. LCMS (ESI) m/z: 372.24 (M+1)+.
[001480] Step 2: Ethyl 9'-(cvclopropylmethoxy)-1 '-fluoro-10'-formyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000304_0001
[001481] Ethyl 1 '-fluoro-10'-formyl-9'-hydroxy-2'-oxo-2', 7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (94 mg, 0.253 mmol) in DMF (6ml_) was added cesium carbonate (247 mg, 0.759 mmol) and
(bromomethyl)cyclopropane (0.05 ml, 0.516 mmol) and heated to 90°C for several hours. The reaction was then cooled to room temperature, diluted with brine and extracted with diethyl ether and ethyl acetate. The combined organics were dried over sodium sulfate and placed under vacuum to give the product (94 mg) that was used without further purification in the next step. LCMS (ESI) m/z: 426.30 (M+1)+.
[001482] Step 3: Ethyl 9'-(cvclopropylmethoxy)-1 Q'-(difluoromethyl)-1 '-fluoro-
Figure imgf000304_0002
[001483] Ethyl 9'-(cyclopropylmethoxy)-1 '-fluoro-10'-formyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (94 mg, 0.221 mmol) was dissolved in DCM and the reaction was cooled to 0 °C. DAST (0.12 ml_, 0.908 mmol) was added dropwise and the reaction was stirred at room temperature until the reaction was complete by LCMS. he reaction was poured into ice water and saturated NaHC03 solution, extracted with EtOAc, the organics were dried over sodium sulfate and concentrated under reduced pressure to give the product (100 mg) that was used without further purification in the next step. LCMS (ESI) m/z: 448.30 (M+1 ).
[001484] Step 4: 9'-(Cvclopropylmethoxy)-10'-(difluoromethyl)-1 '-fluoro-2'-oxo- 2',7'-dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000305_0001
[001485] Ethyl 9'-(cyclopropylmethoxy)-10'-(difluoromethyl)-1 '-fluoro-2'-oxo- 2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (50 mg, 0.1 12 mmol) was dissolved in EtOH (3 mL) and 1 M LiOH (1 1 17 μΙ, 1 .1 17 mmol) was added. The reaction was stirred at room temperature overnight, then concentrated under reduced pressure. The reaction was diluted with water and made acidic with 6 N HCI, then extracted with ethyl acetate. The organices were dried over sodium sulfate and purified by reverse phase HPLC to give white solid (1 .6 mg). Ή NMR (400 MHz, CHLOROFORM-d) δ ppm 0.35 - 0.45 (m, 2 H), 0.65 - 0.74 (m, 2 H), 0.76 - 0.96 (m, 2 H), 1 .92 - 2.19 (m, 4 H), 2.62 (br. s., 2 H), 3.25 (s, 2 H), 3.98 (d, J=7.0 Hz, 2 H), 6.87 (s, 1 H), 8.24 - 8.34 (m, 1 H), 8.85 (s, 1 H); LCMS (ESI) m/z: 420.28 (M+1 )+.
[001486] Example 130
[001487] 10-Methoxy-9-(3-methoxypropoxy)-6,6-dimethyl-2-oxo-6,7-dihydro- 2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000305_0002
[001489] LDA (8.14 ml, 8.14 mmol) was added to a oven dried flask and cooled to 0 °C and ethyl isobutyrate (0.874 ml, 6.51 mmol) was added dropwise. The mixture was stirred at rt for 30 minutes and the mixture was cooled to 0 °C. To this was added a solution of bromide 2-(benzyloxy)-4-(bromomethyl)-1 -methoxybenzene (1 g, 3.26 mmol) in THF (5ml) dropwise and the mixture was warmed to room temperature. The reaction mixture was poured into an ice water / NH4CI solution. The mixture was extracted with EtOAc, washed with brine, dried over sodium sulfate and concentrated in vacuo. The crude material was purified via silica gel chromatography to give 896 mg ethyl 3-(3- (benzyloxy)-4-methoxyphenyl)-2,2-dimethylpropanoate (80% yield). LCMS (ESI) m/z: 365.3 (M+23)+.
[001490] Step 2: 3-(3-(Benzyloxy)-4-methoxyphenyl)-2,2-dimethylpropanoic acid
Figure imgf000306_0001
[001491] Ethyl 3-(3-(benzyloxy)-4-methoxyphenyl)-2,2-dimethylpropanoate (1 g, 2.92 mmol) was dissolved in EtOH (20ml_). LiOH (23.36 ml, 23.36 mmol) (1 .0 M solution) was added, and the reaction mixture was heated at 66 °C overnight. Additional LiOH (2mL) was added and the reaction mixture was heated at 90 °C for 2 hrs. The reaction mixture was concentrated and acidified with 6 N HCI. The aqueous mixture was extracted with EtOAc, dried over sodium sulfate and concentrated to give 790 mg crude 3-(3-(benzyloxy)-4- methoxyphenyl)-2,2-dimethylpropanoic acid. LCMS (ESI) m/z: 313.4 (M - 1)".
[001492] Step 3: 1 -(3-(Benzyloxy)-4-methoxyphenyl)-2-methylpropan-2-amine
Figure imgf000306_0002
[001493] 3-(3-(Benzyloxy)-4-methoxyphenyl)-2,2-dimethylpropanoic acid (567 mg, 1 .804 mmol) and TEA (377 μΙ, 2.71 mmol) were dissolved in t-butanol (2 ml). DPPA (0.5mL, 2.245 mmol) was added and the reaction mixture was heated at 106 °C for 14 hours and at rt for 3 days. The reaction mixture was concentrated and the crude material was dissolved in dioxane (5mL) and treated with 6 N HCI (18 mmol, 3ml) with heating at 107 °C. The reaction mixture was concentrated, diluted with a NaHC03 solution was extracted with EtOAc. The mixture was dried over sodium sulfate and concentrated to give 525 mg crude 1 -(3-(benzyloxy)-4-methoxyphenyl)-2-methylpropan-2-amine which was used as is in the next step. LCMS (ESI) m/z: 286.4 (M+1)+.
[001494] Step 4: N-(1 -(3-(benzyloxy)-4-methoxyphenyl)-2-methylpropan-2- vDformamide
Figure imgf000306_0003
[001495] Crude 1 -(3-(benzyloxy)-4-methoxyphenyl)-2-methylpropan-2-amine (525 mg) was dissolved in dioxane (2 mL) and was treated with TEA (0.513 mL, 3.68 mmol) and acetic formic anhydride (1 .7 mL, 18.40 mmol) (10.8 M solution) for 10 minutes. The reaction mixture was poured into a ice water/ NaHC03 solution and was extracted with EtOAc. The organic layer was dried over sodium sulfate and purified via silica gel chromatography to give 263 mg N-(1 -(3-(benzyloxy)-4-methoxyphenyl)-2-methylpropan-2- yl)formamide. LCMS (ESI) m/z: 314.3 (M+1 )+.
[001496] Step 5: 6-(Benzyloxy)-7-methoxy-3,3-dimethyl-3,4- dihvdroisoquinoline
Figure imgf000307_0001
[001497] POC (138 μΙ, 1 .480 mmol) was added dropwise to a 0 °C solution of N-(1 -(3-(benzyloxy)-4-methoxyphenyl)-2-methylpropan-2-yl)formamide (262 mg, 0.836 mmol) in MeCN (5ml_) and the reaction mixture was heated at 90 °C for 20 min. The reaction mixture was concentrated, diluted with EtOAc/water and made basic with a Nh /MeOH solution. The mixture was extracted with EtOAc, dried over sodium sulfate and concentrated to give 247mg crude 6-(benzyloxy)-7-methoxy-3,3-dimethyl-3,4- dihydroisoquinoline as a yellow oil. LCMS (ESI) m/z: 296.2 (M+1)+.
[001498] Step 6: Ethyl 9-(benzyloxy)-10-methoxy-6,6-dimethyl-2-oxo- 2,6,7,1 1 b-tetrahvdro-1 H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000307_0002
[001499] Crude 6-(benzyloxy)-7-methoxy-3,3-dimethyl-3,4-dihydroisoquinoline was dissolved in EtOH (20 mL). Ethyl 2-(ethoxymethylene)-3-oxobutanoate (218 μΙ, 1 .254 mmol) was added and the mixture was heated at reflux overnight. Additional ethyl 2- (ethoxymethylene)-3-oxobutanoate (218 μΙ, 1 .254 mmol) was added and the reaction mixture was heated for another three hours at reflux. Additional ethyl 2-(ethoxymethylene)- 3-oxobutanoate (218 μΙ, 1 .254 mmol) was added and the mixture was heated at reflux overnight. The reaction mixture was concentrated to give crude ethyl 9-(benzyloxy)-10- methoxy-6,6-dimethyl-2-oxo-2,6,7,1 1 b-tetrahydro-1 H-pyrido[2,1 -a]isoquinoline-3- carboxylate which was used as is in the next step. LCMS (ESI) m/z: 436.3 (M+1)+.
[001500] Step 7: Ethyl 9-(benzyloxy)-10-methoxy-6.6-dimethyl-2-oxo-6.7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000308_0001
[001501] Crude ethyl 9-(benzyloxy)-10-methoxy-6,6-dimethyl-2-oxo-2,6,7,1 1 b- tetrahydro-1 H-pyrido[2,1 -a]isoquinoline-3-carboxylate was dissolved in DME (10 mL), DDQ (190 mg, 0.836 mmol) was added and the reaction mixture was stirred at 85 °C. Additional DDQ (95 mg) was added and the reaction mixture was heated at 85 °C for another hour. The mixture was poured into an ice water/ NaHC03 solution, and the mixture was extracted with EtOAc. The organic layer was dried over sodium sulfate and purified via silica gel chromatography to give 145 mg ethyl 9-(benzyloxy)-10-methoxy-6,6-dimethyl-2-oxo-6,7- dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate. LCMS (ESI) m/z: 434.3 (M+1 )+.
[001502] Step 8: Ethyl 9-hvdroxy-10-methoxy-6,6-dimethyl-2-oxo-6,7-dihvdro- 2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000308_0002
[001503] To ethyl 9-(benzyloxy)-10-methoxy-6,6-dimethyl-2-oxo-6,7-dihydro- 2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (1 12 mg, 0.258 mmol) in 1 :1 THF/MeOH was added Pd/C Degussa type 50% water and purged with 3 balloons of hydrogen at 1 atm. The reaction mixture was filtered and was concentrated to give 77 mg ethyl 9-hydroxy-10- methoxy-6,6-dimethyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate which was used directly in the next step. LCMS (ESI) m/z: 344.4 (M+1)+.
[001504] Step 9: Ethyl 10-methoxy-9-(3-methoxypropoxy)-6,6-dimethyl-2-oxo- 6,7-dihvdro-2H-pyrido[ -alisoquinoline-3-carboxylate
Figure imgf000308_0003
[001505] Ethyl 9-hydroxy-10-methoxy-6,6-dimethyl-2-oxo-6,7-dihydro-2H- pyrido[2,1 -a]isoquinoline-3-carboxylate (77 mg, 0.224 mmol), 1 -bromo-3-methoxypropane (54 mg, 0.356 mmol) and cesium carbonate (95 mg, 0.292 mmol) in DMF was stirred at rt overnight. The reaction mixture was diluted with Et20 and water. The organic layer was separated, washed with brine, dried over sodium sulfate and purified via silica gel chromatography to give 61 mg ethyl 10-methoxy-9-(3-methoxypropoxy)-6,6-dimethyl-2-oxo- 6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (66 % yield). LCMS (ESI) m/z: 416.3 (M+1)+.
[001506] Step 10: 10-Methoxy-9-(3-methoxypropoxy)-6,6-dimethyl-2-oxo-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000309_0001
[001507] Ethyl 10-methoxy-9-(3-methoxypropoxy)-6,6-dimethyl-2-oxo-6,7- dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (60 mg, 0.144 mmol) dissolved in EtOH was treated with 1 N NaOH (2.1 ml_, 2.16 mmol) for 20 minutes. The reaction mixture was concentrated and acidified with 6N HCI. The aqueous mixture was extracted with EtOAc, dried over sodium sulfate and was purified via reverse phase HPLC to give 34 mg 10- Methoxy-9-(3-methoxypropoxy)-6,6-dimethyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 - a]isoquinoline-3-carboxylic acid. 1 H NMR (400 MHz, CHLOROFORM-d) δ ppm 1 .59 (s, 6 H), 2.15 (t, J=6.1 Hz, 2 H), 3.01 (s, 2 H), 3.36 (s, 3 H), 3.58 (t, J=6.1 Hz, 2 H), 3.93 (s, 3 H), 4.18 (t, J=6.4 Hz, 2 H), 6.75 (s, 1 H), 7.09 (s, 1 H), 7.18 (s, 1 H), 8.78 (s, 1 H). LCMS (ESI) m/z: 388.30 (M+1)+.
[001508] Example 131
[001509] 10'-Methoxy-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihydrospirofcyclopropane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000309_0002
[001510] -(4-Methoxy-3-(3-methoxypropoxy)phenyl)acetonitrile
Figure imgf000309_0003
[001511] 4-(Bromomethyl)-1 -methoxy-2-(3-methoxypropoxy)benzene (546 mg, 1 .89 mmol) was dissolved in DMF (4 ml_) and added NaCN (213 mg, 4.34 mmol) and the reaction mixture was stirred overnight at room temperature. The reaction was diluted with diethyl ether and water. The organic layer was washed with brine, the aqueous layer was back extracted x4, and the organics were dried over sodiums suflate. The crude material was purified using silica-gel chromatography(0-100% EtOAc/hexanes gradient elution) to give the product as a colorless oil (397 mg, 89%). LCMS (ESI) m/z: 236.30 (M+1)+.
Figure imgf000310_0001
[001512] Step 2: 1 -(4-Methoxy-3-(3- methoxypropoxy)phenyl)cvclopropanamine
[001513] To 2-(4-Methoxy-3-(3-methoxypropoxy)phenyl)acetonitrile (387 mg,
1 .645 mmol) in diethyl ether (5ml_) was added titanium(IV) isopropoxide (0.52 ml_, 1 .775 mmol) and cooled to 0 °C. Ethyl magnesium bromide solution (1 .15 ml_, 3.45 mmol) was added and the brown suspension was stirred 30 minutes at 0 °C. Boron trifluoride etherate (0.44 ml_, 3.47 mmol) was added and the reaction was stirred at 0 °C for 15 minutes and then at room temperature overnight. The reaction was poured into ice and 1 N NaOH, diluted with ethyl acetate and the biphasic suspension was stirred vigorously for 1 hour, filtered and the the organic layer was separated. The organics were dried over sodium sulfate and concentrated under reduced pressure to give desired product impure with unreacted starting material. The combined aqueous layers were extracted again with a mixture of chloroform: IPA (3:1) and the combined organics from this extraction were dried over sodium sulfate and concentrated under reduced pressure to give clean desired product (30 mg, 7%) which was used without further purification in the next steps. LCMS (ESI) m/z: 266.30 (M+1)+.
[001514] Step 3: N-(1 -(4-methoxy-3-(3- methoxypropoxy)benzyl)cvclopropyl)formamide
Figure imgf000310_0002
[001515] To a mixture of 1 -(4-methoxy-3-(3- methoxypropoxy)benzyl)cyclopropan-1 -amine (30 mg, 0.1 13 mmol) in dioxane was added TEA (0.032 ml_, 0.226 mmol), formic acetic anhydride (0.1 ml_, 0.679 mmol) and the reaction mixture was stirred at rt for 10 minutes. The reaction mixture was poured into an ice water/NaHC03 solution and was extracted with EtOAc. The organic layer was dried over sodium sulfate and concentrated to give crude N-(1 -(4-methoxy-3-(3- methoxypropoxy)benzyl)cyclopropyl)formamide which was used in the next step as is. LCMS (ESI) m/z: 294.3 (M+1)+.
[001516] Step 4: 7'-Methoxy-6'-(3-methoxypropoxy)-4'H-spiro[cvclopropane- 1 ,3'-isoquinolinel
Figure imgf000311_0001
[001517] Crude N-(1 -(4-methoxy-3-(3- methoxypropoxy)benzyl)cyclopropyl)formamide was dissolved in MeCN and cooled to 0 °C. POCI3 (0.019 mL, 0.200 mmol) was added dropwise and the reaction was heated at 90 °C for 15 minutes. The reaction mixture was concentrated, diluted with ice water/NaHC03 solution and was extracted with EtOAc. The organic layer was dried over sodium sulfate and concentrated to give crude 7'-methoxy-6'-(3-methoxypropoxy)-4'H-spiro[cyclopropane- 1 ,3'-isoquinoline] which was used directly in the next step. LCMS (ESI) m/z: 276.2 (M+1)+.
[001518] Step 5: Ethyl 10-methoxy-9-(3-methoxypropoxy)-6,6-dimethyl-2-oxo- 6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000311_0002
[001519] Crude 7'-methoxy-6'-(3-methoxypropoxy)-4'H-spiro[cyclopropane- 1 ,3'-isoquinoline] from previous step was dissolved in EtOH, ethyl 2-(ethoxymethylene)-3- oxobutanoate (0.1 mL, 0.575 mmol) was added and the mixture was stirred with heating at reflux overnight. The reflux condenser was removed and heating was continued until mixture was concentrated. Additional 0.45 mL of ethyl 2-(ethoxymethylene)-3-oxobutanoate was added and the mixture was stirred for another 30 minutes under reflux. The reaction mixture was concentrated, the crude material was dissolved in DME, DDQ (38.5 mg, 0.170 mmol) was added and the reaction mixture was heated with stirring at 80 °C for 30 minutes. The reaction mixture was poured into an ice water/NaHC03 solution and was extracted with EtOAc. The organic layer was dried over sodium sulfate and concentrated, and the crude material was purified via silica gel chromatography to give 10.6 mg of ethyl 10-methoxy-9- (3-methoxypropoxy)-6,6-dimethyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3- carboxylate. LCMS (ESI) m/z: 414.3 (M+1) +.
[001520] Step 6: 10'-Methoxy-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihydrospirofcyclopropane-1 ,6'-pyrido[2,1 -al isoquinolinel-3'-carboxylic acid
Figure imgf000311_0003
[001521] To a solution of ethyl 10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo- 2',7'-dihydrospiro[cyclopropane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (10.6 mg, 0.026 mmol) in EtOH (2ml_) was added 1 N NaOH (0.6 mL, 0.600 mmol) and the reaction mixture was stirred overnight at room temperature. The reaction mixture was concentrated, diluted with water, acidified and extracted. The crude material was purified to give 4 mg of 10'-Methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclopropane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylic acid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 1 .08 (s, 2 H), 1 .44 (s, 2 H), 2.14 (t, J=6.2 Hz, 2 H), 2.94 (s, 2 H), 3.36 (s, 3 H), 3.58 (t, J=5.9 Hz, 2 H), 3.94 (s, 3 H), 4.18 (t, J=6.4 Hz, 2 H), 6.74 (s, 1 H), 7.07 (s, 1 H), 7.20 (s, 1 H), 8.48 (s, 1 H); LCMS (ESI) m/z: 386.3 (M+1)+.
[001522] Example 132
Figure imgf000312_0001
[001525] n-Butyllithium (2.5 M in hexanes) (1 .24 mL, 3.09 mmol) was added dropwise to a -78 °C solution of diisopropylamine (0.440 mL, 3.09 mmol) in tetrahydrofuran (THF) (13 mL). The mixture was allowed to warm to room temperature and stirred for 15 minutes. The mixture was cooled to -78 °C before methyl cyclopentanecarboxylate (0.355 mL, 2.81 mmol) was added dropwise. The mixture was warmed to 0 °C and stirred for 30 minutes before being cooled again to -78 °C. A solution of 2-(benzyloxy)-4-(bromomethyl)- 1 -methoxybenzene (1 .035 g, 3.37 mmol) in tetrahydrofuran (THF) (5 mL) was added dropwise. The mixture was stirred an additional 15 minutes at -78 °C and then allowed to warm to room temperature and stirred overnight. The mixture was diluted with ethyl acetate, washed with saturated ammonium chloride, washed with brine, and concentrated. The residue was purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient). Fractions were
concentrated. The residue was dissolved in acetonitrile and the mixture concentrated in order to azeotrope remaining water. Drying under vacuum gave methyl 1 -(3-(benzyloxy)-4- methoxybenzyl)cyclopentane-1 -carboxylate (716 mg, 2.02 mmol, 72 % yield) as a clear oil. LCMS (ESI) m/z: 377.3 (M+Na)+.
[001526] Step 2: 1 -(3-(Benzyloxy)-4-methoxybenzyl)cvclopentanecarboxylic acid
Figure imgf000313_0001
[001527] 1 M Sodium hydroxide (10 ml_, 10 mmol) was added to a solution of methyl 1 -(3-(benzyloxy)-4-methoxybenzyl)cyclopentane-1 -carboxylate (710 mg, 2.00 mmol) in methanol (10 ml_) and tetrahydrofuran (THF) (10 ml_). The mixture was heated at 60 °C for 2 hours, cooled to room temperature, and concentrated to ~10 ml_. The mixture was acidified with 1 M hydrochloric acid and extracted 3 times with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give 1 -(3-(benzyloxy)-4-methoxybenzyl)cyclopentane-1 -carboxylic acid (61 1 mg, 1 .80 mmol, 90 % yield). LCMS (ESI) m/z: 339.3 (M-1)".
[001528] Step 3: 1 -(3-(Benzyloxy)-4-methoxybenzyl)cvclopentanamine
Figure imgf000313_0002
[001529] Diphenyl phosphorazidate (0.503 ml_, 2.333 mmol) was added to a solution of 1 -(3-(benzyloxy)-4-methoxybenzyl)cyclopentane-1 -carboxylic acid (61 1 mg, 1 .80 mmol) and triethylamine (0.325 ml_, 2.33 mmol) in tert-butanol (7 ml_). The mixture was heated at 85 °C over the weekend. The mixture was concentrated and the residue dissolved in tetrahydrofuran. Hydrogen chloride (4M in dioxane) (5 ml_, 20 mmol) was added and the mixture stirred at room temperature for 2.5 hours. The mixture was concentrated and the residue purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient). Fractions were combined and diluted with 1 M sodium hydroxide and brine. The mixture was extracted 3 times with dichloromethane. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give 1 -(3-(benzyloxy)-4- methoxybenzyl)cyclopentan-1 -amine (360 mg, 1 .16 mmol, 64.4 % yield) as an oil. LCMS (ESI) m/z: 312.3 (M+1)+.
[001530] Step 4: N-(1 -(3-(Benzyloxy)-4-methoxybenzyl)cvclopentyl)formamide
Figure imgf000314_0001
[001531] 1 -(3-(Benzyloxy)-4-methoxybenzyl)cyclopentan-1 -amine (360 mg,
1 .13 mmol) and formic acid (0.217 mL, 5.66 mmol) in 1 ,4-dioxane (3 mL) were heated at
100 °C overnight. The mixture was allowed to cool to room temperature and concentrated to give crude N-(1 -(3-(benzyloxy)-4-methoxybenzyl)cyclopentyl)formamide (385 mg). LCMS
(ESI) m/z: 340.3 (M+1)+.
[001532] Step 5: 6'-(Benzyloxy)-7'-methoxy-4'H-spiro[cvclopentane-1 ,3'- isoquinolinel
Figure imgf000314_0002
[001533] N-(1 -(3-(Benzyloxy)-4-methoxybenzyl)cyclopentyl)formamide (385 mg, 1 .13 mmol) in acetonitrile (3 mL) were cooled in an ice bath before phosphorus oxychloride (0.127 mL, 1 .36 mmol) was added dropwise with stirring. The mixture was heated at 60 °C for 4 hours and concentrated. The residue was dissolved in ethyl acetate and cooled in an ice bath before water and 30% NH4OH were added to bring the pH =~1 1 .
The organic layer was collected and the aqueous layer extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give crude 6'-(benzyloxy)-7'-methoxy-4'H-spiro[cyclopentane-1 ,3'- isoquinoline] (320 mg). LCMS (ESI) m/z: 322.3 (M+1)+.
[001534] Step 6: Ethyl 9'-(benzyloxy)-10'-methoxy^'-oxo^'J1- dihydrospirofcyclopentane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000314_0003
[001535] 6'-(Benzyloxy)-7'-methoxy-4'H-spiro[cyclopentane-1 ,3'-isoquinoline] (300 mg, 0.933 mmol) and ethyl 2-(ethoxymethylene)-3-oxobutanoate (0.487 mL, 2.80 mmol) in ethanol (3 mL) were heated at reflux overnight. Additional ethyl 2- (ethoxymethylene)-3-oxobutanoate (0.487 mL, 2.80 mmol) was added and the mixture continued to heat at 80 °C overnight. The mixture was concentrated and the dark red residue dissolved in 1 ,2-dimethoxyethane (DME) (3 mL). The mixture was heated at 85 °C for 2 hours. The mixture was allowed to cool to room temperature and then cooled in an ice bath. Solids were collected by filtration, washed with cold DME, and dried to give ethyl 9'- (benzyloxy)-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclopentane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (137 mg, 0.298 mmol, 32 % yield). LCMS (ESI) m/z: 460.3 (M+1)+. The filtrate was concentrated and the residue purified by silica chromatography eluting with a gradient of 0% to 50% ethanol in ethyl acetate. Fractions were concentrated and the residue sonicated with ethyl acetate. Ethyl ether was added and the mixture sonicated again. Solids were collected by filtration and dried to give ethyl 9'-(benzyloxy)- 10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclopentane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (125 mg, 0.272 mmol, 29.1 % yield) as a brown solid. LCMS (ESI) m/z: 460.3 (M+1)+.
[001536] Step 7: 9'-(Benzyloxy)-10'-methoxy-2'-oxo-2'.7'- dihydrospirofcyclopentane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000315_0001
[001537] A solution of lithium hydroxide monohydrate (1 1 .4 mg, 0.272 mmol) in water (0.25 mL) was added to a stirring solution of ethyl 9'-(benzyloxy)-10'-methoxy-2'- oxo-2', 7'-dihydrospiro[cyclopentane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (25 mg, 0.054 mmol) in methanol (0.75 mL) and the mixture stirred overnight at room temperature. The mixture was diluted with water, acidified with 1 M hydrochloric acid, and extracted 3 times with dichloromethane. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give 9'-(benzyloxy)-10'-methoxy-2'-oxo-2',7'- dihydrospiro[cyclopentane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (22 mg, 0.050 mmol, 93 % yield) as a powder. Ή NMR (400 MHz, DMSO-c/6) δ ppm 1 .60 - 1 .93 (m, 6 H) 2.01 - 2.15 (m, 2 H) 3.14 (s, 2 H) 3.87 (s, 3 H) 5.15 (s, 2 H) 7.1 1 (s, 1 H) 7.31 - 7.55 (m, 7 H) 8.50 (s, 1 H). 24 of 25 protons found. LCMS (ESI) m/z: 432.3 (M+1)+.
[001538] Example 133
[001539] 10'-Methoxy-9'-(3-methoxypropoxy)-2'-oxo-2'.7'- dihydrospirofcyclopentane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000315_0002
Figure imgf000316_0001
[001541] A round bottom flask with stir bar, ethyl 9'-(benzyloxy)-10'-methoxy-
2'-oxo-2',7'-dihydrospiro[cyclopentane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (260 mg, 0.566 mmol) and 10% palladium on carbon (40 mg, 0.038 mmol) in tetrahydrofuran
(THF) (2 mL) and methanol (2 mL) was first purged with nitrogen and then with hydrogen.
The mixture was hydrogenated under balloon pressure for 4 hours. The mixture was quenched with a stream of nitrogen before being filtered over celite. The filtrate was concentrated to give ethyl 9'-hydroxy-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclopentane-
1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (193 mg, 0.522 mmol, 92 % yield) as a pale mustard solid. LCMS (ESI) m/z: 370.4 (M+1)+.
[001542] Step 2: Ethyl 10'-methoxy-9'-(3-methoxypropoxyV2'-oxo-2'.7'- dihydrospirofcyclopentane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000316_0002
[001543] Ethyl 9'-hydroxy-10'-methoxy-2'-oxo-2',7'-dihydrospiro[cyclopentane- 1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (107 mg, 0.290 mmol), potassium carbonate (120 mg, 0.869 mmol), and 1 -bromo-3-methoxypropane (89 mg, 0.579 mmol) in N,N- dimethylformamide (DMF) (2 mL) were stirred at room temperature overnight. The mixture was quenched with water and extracted 2 times with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give ethyl 10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclopentane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (128 mg, 0.290 mmol, 100 % yield). LCMS (ESI) m/z: 442.3 (M+1)+.
[001544] Step 3: 10'-Methoxy-9'-(3-methoxypropoxyV2'-oxo-2'.7'- dihydrospirofcyclopentane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000317_0001
[001545] A solution of lithium hydroxide monohydrate (73.0 mg, 1 .74 mmol) in water (0.68 mL) was added to a stirring solution of ethyl 10'-methoxy-9'-(3- methoxypropoxy)-2'-oxo-2',7'-dihydrospiro[cyclopentane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (128 mg, 0.290 mmol) in methanol (2 mL) and the mixture stirred at 60 °C for 3 hours. The mixture was allowed to cool to room temperature, diluted with water, acidified with 1 M hydrochloric acid, and extracted 3 times with dichloromethane. The combined organic layers were washed with brine and concentrated. The residue was purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient). Fractions were concentrated. The residue was lyophililzed (acetonitrile / water) to give 10'-methoxy-9'-(3-methoxypropoxy)-2'-oxo-2',7'- dihydrospiro[cyclopentane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (49 mg, 0.1 19 mmol, 41 % yield) as a pale tan solid. 1H NMR (400 MHz, DMSO-cf6) δ ppm 1 .64 - 1 .74 (m, 2 H), 1 .75 - 1 .91 (m, 4 H), 1 .97 (quin, J=6.25 Hz, 2 H), 2.02 - 2.13 (m, 2 H), 3.14 (s, 2 H), 3.23 (s, 3 H), 3.46 (t, J=6.25 Hz, 2 H), 3.86 (s, 3 H), 4.08 (t, J=6.44 Hz, 2 H), 6.99 (s, 1 H), 7.47 (s, 1 H), 7.49 (s, 1 H), 8.49 (s, 1 H). 26 of 27 protons found. LCMS (ESI) m/z: 414.3 (M+1)+.
[001546] Example 134
Figure imgf000317_0002
[001549] n-Butyllithium (2.5 M in hexanes) (1 .72 mL, 4.30 mmol) was added dropwise to a -78 °C solution of diisopropylamine (0.612 mL, 4.30 mmol) in tetrahydrofuran (THF) (13 mL). The mixture was allowed to warm to room temperature and stirred for 15 minutes. The mixture was cooled to -78 °C before methyl 3,3-dimethylcyclobutane-1 - carboxylate (0.555 g, 3.91 mmol) was added dropwise. The mixture was warmed to 0 °C and stirred for 30 minutes before being cooled again to -78 °C. A solution of 2-(benzyloxy)-
4-(bromomethyl)-1 -methoxybenzene (1 .2 g, 3.9 mmol) in tetrahydrofuran (THF) (5 mL) was added dropwise. The mixture was stirred an additional 15 minutes at -78 °C and then allowed to warm to room temperature and stirred overnight. The mixture was diluted with ethyl acetate, washed with saturated ammonium chloride, washed with brine, and concentrated. The residue was purified by medium pressure reverse phase
chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient).
Fractions were concentrated. The residue was dissolved in acetonitrile and the mixture concentrated in order to azeotrope remaining water. Drying under vacuum gave methyl 1 -
(3-(benzyloxy)-4-methoxybenzyl)-3,3-dimethylcyclobutane-1 -carboxylate (380 mg, 1 .03 mmol, 26 % yield). LCMS (ESI) m/z: 391 .4 (M+Na)+. A second set of fractions from the same peak was concentrated. The residue was dissolved in acetonitrile and the mixture concentrated in order to azeotrope remaining water. Drying under vacuum gave methyl 1 -
(3-(benzyloxy)-4-methoxybenzyl)-3,3-dimethylcyclobutane-1 -carboxylate (456 mg, 1 .238 mmol, 32 % yield). LCMS (ESI) m/z: 391 .4 (M+Na)+. 58% overall yield.
Figure imgf000318_0001
[001551] 1 M Sodium hydroxide (20 mL, 20 mmol) was added to a solution of methyl 1 -(3-(benzyloxy)-4-methoxybenzyl)-3,3-dimethylcyclobutane-1 -carboxylate (836 mg,
2.269 mmol) in methanol (10 mL) and tetrahydrofuran (THF) (10 mL). The mixture was heated at 80 °C for 2 hours, cooled to room temperature, and concentrated to ~10 mL. The mixture was acidified with 1 M hydrochloric acid and extracted 3 times with dichloromethane.
The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give 1 -(3-(benzyloxy)-4-methoxybenzyl)-3,3-dimethylcyclobutane-1 - carboxylic acid (757 mg, 2.14 mmol, 94 % yield). LCMS (ESI) m/z: 353.4 (M-1)".
[001552] Step 3: 1 -(3-(Benzyloxy)-4-methoxybenzyl)-3,3- dimethylcvclobutanami
Figure imgf000318_0002
[001553] Diphenyl phosphorazidate (0.593 mL, 2.75 mmol) was added to a solution of 1 -(3-(benzyloxy)-4-methoxybenzyl)-3,3-dimethylcyclobutane-1 -carboxylic acid (750 mg, 2.12 mmol) and triethylamine (0.383 mL, 2.75 mmol) in tert-butanol (5 mL). The mixture was heated at 85 °C overnight and concentrated. The residue was dissolved in dichloromethane before hydrogen chloride (4M in dioxane) (5 mL, 20 mmol) was added. The mixture was stirred for 4 hours and concentrated. The residue was purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / formic acid / 10% to 100% gradient). Combined fractions were concentrated to remove most the acetonitrile. The remaining aqueous portion was made basic with 1 M sodium hydroxide and extracted 3 times with dichloromethane. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give 1 -(3-(benzyloxy)-4-methoxybenzyl)-3,3- dimethylcyclobutan-1 -amine (205 mg, 0.630 mmol, 30 % yield). LCMS (ESI) m/z: 326.3 (M+1)+.
[001554] Step 4: N-(1 -(3-(Benzyloxy)-4-methoxybenzyl)-3.3- dimethylcvclobutvDformamide
Figure imgf000319_0001
[001555] 1 -(3-(Benzyloxy)-4-methoxybenzyl)-3,3-dimethylcyclobutan-1 -amine (205 mg, 0.617 mmol) and formic acid (0.237 mL, 6.17 mmol) in 1 ,4-dioxane (3 mL) were heated at 100 °C overnight. The mixture was allowed to cool to room temperature and concentrated to give N-(1 -(3-(benzyloxy)-4-methoxybenzyl)-3,3- dimethylcyclobutyl)formamide (218 mg, 0.617 mmol, 100 % yield). LCMS (ESI) m/z: 354.3 (M+1)+.
Figure imgf000319_0002
[001557] N-(1 -(3-(Benzyloxy)-4-methoxybenzyl)-3,3- dimethylcyclobutyl)formamide (218 mg, 0.617 mmol) in acetonitrile (2 mL) were cooled in an ice bath before phosphorus oxychloride (0.069 mL, 0.74 mmol) was added dropwise with stirring. The mixture was heated at 60 °C for 4 hours and concentrated. The residue was dissolved in ethyl acetate and cooled in an ice bath before water and 30% NH4OH were added to bring the pH =~1 1 . The organic layer was collected and the aqueous layer extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give 6'-(benzyloxy)-7'-methoxy-3,3-dimethyl-4'H- spiro[cyclobutane-1 ,3'-isoquinoline] (207 mg, 0.617 mmol, 100 % yield). LCMS (ESI) m/z: 336.3 (M+1)+.
Figure imgf000320_0001
[001559] 6'-(Benzyloxy)-7'-methoxy-3,3-dimethyl-4'H-spiro[cyclobutane-1 ,3'- isoquinoline] (207 mg, 0.617 mmol) and ethyl 2-(ethoxymethylene)-3-oxobutanoate (0.537 ml_, 3.09 mmol) in ethanol (3 ml_) were heated at reflux overnight. The mixture was concentrated and the residue dissolved in 1 ,2-dimethoxyethane (DME) (3 ml_). p-Chloranil
(152 mg, 0.617 mmol) was added and the mixture heated at 85 °C for 2 hours. The mixture was cooled in an ice bath. The precipitate was collected by filtration, washed with cold
DME, and dried to give ethyl 9'-(benzyloxy)-10'-methoxy-3,3-dimethyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (96 mg, 0.20 mmol,
33 % yield) as a light tan solid. LCMS (ESI) m/z: 473.3 (M+1 )+. The filtrate was
concentrated and the residue purified by silica chromatography eluting with a gradient of 0% to 100% (25% EtOH / EtOAc) in hexanes. Fractions were concentrated to give ethyl 9'-
(benzyloxy)-10'-methoxy-3,3-dimethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (95 mg, 0.20 mmol, 33 % yield). LCMS (ESI) m/z: 474.4
(M+1)+. 66% overall yield.
[001560] Step 7: 9'-(Benzyloxy)-10'-methoxy-3.3-dimethyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000320_0002
[001561] A solution of lithium hydroxide monohydrate (22.2 mg, 0.528 mmol) in water (0.25 ml_) was added to a solution of ethyl 9'-(benzyloxy)-10'-methoxy-3,3-dimethyl- 2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (25 mg, 0.053 mmol) in methanol (0.75 ml_) and the mixture heated at 60 °C for 3 hours. The mixture was allowed to cool to room temperature, diluted with water, acidified with 1 M hydrochloric acid, and extracted 3 times with dichloromethane. The combined organic layers were washed with brine and concentrated. The residue was purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient). Fractions were concentrated. The residue was lyophililzed (acetonitrile / water) to give 9'-(benzyloxy)-10'-methoxy-3,3-dimethyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (20 mg, 0.045 mmol, 85 % yield). Ή NMR (400 MHz, DMSO-c/6) δ ppm 1 .18 (s, 3 H), 1 .23 (s, 3 H), 2.06 (d, J=14.44 Hz, 2 H), 2.43 (d, J=14.05 Hz, 2 H), 3.27 (s, 2 H), 3.86 (s, 3 H), 5.17 (s, 2 H), 7.23 (s, 1 H), 7.31 - 7.37 (m, 1 H), 7.40 (t, J=7.22 Hz, 2 H), 7.44 - 7.51 (m, 4 H), 8.84 (s, 1 H). 26 of 27 protons found. LCMS (ESI) m/z: 446.3(M+1)+.
[001562] Example 135
[001563] 10'-Methoxy-9'-(3-methoxypropoxyV3.3-dimethyl-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000321_0001
[001564] Step 1 : Ethyl 9'-hvdroxy-10'-methoxy-3.3-dimethyl-2'-oxo-2'.7'- dihvdrospiro[cvclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000321_0002
[001565] A reaction vial containing a stir bar, ethyl 9'-(benzyloxy)-10'-methoxy- 3,3-dimethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'- carboxylate (162 mg, 0.342 mmol), 10% palladium on carbon (35 mg, 0.033 mmol), methanol (2 ml_), and tetrahydrofuran (THF) (2 ml_) was purged with nitrogen and then hydrogen. The mixture was hydrogenated for 3 hours under balloon pressure. The mixture was quenched a stream of nitrogen and filtered over celite. The filtrate was concentrated to give ethyl 9'-hydroxy-10'-methoxy-3,3-dimethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'- pyrido[2,1 -a]isoquinoline]-3'-carboxylate (131 mg, 0.342 mmol, 100 % yield). LCMS (ESI) m/z: 384.3 (M+1)+.
Figure imgf000321_0003
[001567] Ethyl 9'-hydroxy-10'-methoxy-3,3-dimethyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (102 mg, 0.266 mmol), potassium carbonate (1 10 mg, 0.798 mmol), and 1 -bromo-3-methoxypropane (0.060 ml_, 0.532 mmol) in Ν,Ν-dimethylformamide (DMF) (2 ml_) were stirred at room temperature overnight. The mixture was quenched with brine and extracted 2 times with ethyl acetate.
The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give ethyl 10'-methoxy-9'-(3-methoxypropoxy)-3,3-dimethyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (1 15 mg, 0.252 mmol, 95 % yield). LCMS (ESI) m/z: 456.4 (M+1 )+.
[001568] Step 3: 10'-Methoxy-9'-(3-methoxypropoxy)-3.3-dimethyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000322_0001
[001569] A solution of lithium hydroxide monohydrate (63.6 mg, 1 .52 mmol) in water (0.68 ml_) was added to a stirring solution of ethyl 10'-methoxy-9'-(3- methoxypropoxy)-3,3-dimethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (1 15 mg, 0.252 mmol) in methanol and the mixture stirred at 60 °C for 3 hours. The mixture was allowed to cool to room temperature, diluted with water, acidified with 1 M hydrochloric acid, and extracted 3 times with dichloromethane. The combined organic layers were washed with brine and concentrated. The residue was purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient). Fractions were concentrated. The residue was lyophililzed (acetonitrile / water) to give 10'-methoxy-9'-(3-methoxypropoxy)-3,3-dimethyl-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (70 mg, 0.16 mmol, 65 % yield) as an off-white solid. Ή NMR (400 MHz, DMSO-cf6) δ ppm 1 .17 (s, 3 H), 1 .24 (s, 3 H), 1 .89 - 2.02 (m, 2 H), 2.03 - 2.12 (m, 2 H), 2.42 (d, J=14.05 Hz, 2 H), 3.30 (d, J=16.78 Hz, 5 H), 3.47 (t, J=6.25 Hz, 2 H), 3.86 (s, 3 H), 4.10 (t, J=6.44 Hz, 2 H), 7.12 (s, 1 H), 7.43 (s, 1 H), 7.46 (s, 1 H), 8.84 (s, 1 H). 28 of 29 protons found. LCMS (ESI) m/z: 428.3 (M+1)+.
[001570] Example 136
[001571] 10'-Borono-9'-(3-methoxypropoxy)-3,3-dimethyl-2'-oxo-2',7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000323_0001
[001573] A reaction vial containing a stir bar, ethyl 10'-chloro-9'-(3- methoxypropoxy)-3,3-dimethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 - a]isoquinoline]-3'-carboxylate (94 mg, 0.21 mmol), potassium acetate (60.2 mg, 0.613 mmol), bis(pinacolato)diboron (78 mg, 0.31 mmol), tris(dibenzylideneacetone)dipalladium(0) (10 mg, 10.9 μηιοΙ), and X-Phos (20 mg, 0.042 mmol) was purged with nitrogen for 20 minutes before 1 ,4-dioxane (2 ml_) was added and the reaction vial placed into a heating block that was preheated to 95 °C. The mixture was stirred and heated for 4 hours, cooled to room temperature, and filtered through a pad of celite. The celite pad was washed with 3 mL of 1 ,4-dioxane so that the final volume of filtrate was ~5 ml_. 32% hydrochloric acid (2 ml_, 21 .07 mmol) was added and the mixture heated overnight at 85 °C. The mixture was allowed to cool to room temperature and concentrated. The residue was purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient). Fractions were concentrated and the residue lyophilized to give 10'-borono-9'-(3-methoxypropoxy)-3,3-dimethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (10 mg, 0.022 mmol, 1 1 % yield) as a white solid. Ή NMR (400 MHz, DMSO-c/6) δ ppm 8.83 (s, 1 H), 8.06 (s, 1 H), 7.91 (s, 2 H), 7.19 (s, 1 H), 7.13 (s, 1 H), 4.15 (t, J=6.05 Hz, 2 H), 3.50 (t, J=5.86 Hz, 2 H), 3.35 (br. s., 2 H), 3.23 (s, 3 H), 1 .93 - 2.13 (m, 4 H), 1 .23 (s, 3 H), 1 .16 (s, 3 H). 25 of 28 protons found. LCMS (ESI) m/z: 442.3 (M+1)+.
[001574] Example 137
[001575] 10"-Methoxy-9"-(3-methoxypropoxy)-2"-oxo-2".7"- dihydrodispirofcyclobutane-1 ,1 '-cvclobutane-3',6"-pyrido[2,1 -alisoquinolinel-3"-carboxylic acid
Figure imgf000324_0001
[001577] n-Butyllithium (2.5 M in hexanes) (2.32 ml_, 5.81 mmol) was added dropwise to a -78 °C solution of diisopropylamine (0.828 ml_, 5.81 mmol) in tetrahydrofuran
(THF) (20 ml_). The mixture was allowed to warm to room temperature and stirred for 15 minutes. The mixture was cooled to -78 °C before ethyl spiro[3.3]heptane-2-carboxylate
(931 mg, 5.53 mmol) was added dropwise. The mixture was warmed to 0 °C and stirred for
15 minutes before a solution of 4-(bromomethyl)-1 -methoxy-2-(3-methoxypropoxy)benzene
(800 mg, 2.77 mmol) in tetrahydrofuran (THF) (7 ml_) was added dropwise. The mixture was stirred an additional 20 minutes at 0 °C and then quenched with saturated ammonium chloride. The mixture was extracted 2 times with ethyl acetate. The combined organic layers were washed with brine, dried over sodium suflate, and concentrated to give crude ethyl 2-(4-methoxy-3-(3-methoxypropoxy)benzyl)spiro[3.3]heptane-2-carboxylate (1042 mg). LCMS (ESI) m/z: 377.3 (M+1 )+.
[001578] Step 2: 2-(4-Methoxy-3-(3- methoxypropoxy)benzyl)spiro[3.31heptane-2-carboxylic acid
Figure imgf000324_0002
[001579] A solution of lithium hydroxide monohydrate (1 161 mg, 27.7 mmol) in water (5 ml_) was added to a solution of ethyl 2-(4-methoxy-3-(3- methoxypropoxy)benzyl)spiro[3.3]heptane-2-carboxylate (1042 mg, 2.77 mmol) in methanol (5 ml_) and tetrahydrofuran (THF) (5 ml_). The mixture was stirred at room temperature overnight. The mixture was heated at 60 °C for 24 hours and allowed to cool to room temperature. The mixture was concentrated to ~ 5 ml_, acidified with 1 N hydrochloric acid, and extracted 3 times with dichloromethane. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated. The residue was purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient). Fractions were concentrated and the residue lyophilized (AcCN / water) to give 2-(4-methoxy-3-(3-methoxypropoxy)benzyl)spiro[3.3]heptane-2- carboxylic acid (667 mg, 1 .914 mmol, 69.2 % yield) as a white powder. LCMS (ESI) m/z: 347.3 (M-1)-.
[001580] Step 3: 2-(4-Methoxy-3-(3-methoxypropoxy)benzyl)spiro[3.31heptan- 2-amine
Figure imgf000325_0001
[001581] Diphenyl phosphorazidate (0.495 ml_, 2.30 mmol) was added dropwise to a 0 °C stirring mixture of 2-(4-methoxy-3-(3- methoxypropoxy)benzyl)spiro[3.3]heptane-2-carboxylic acid (667 mg, 1 .91 mmol) and triethylamine (0.320 ml_, 2.30 mmol) in toluene (12 ml_). The mixture was allowed to warm to room temperature and stirred for 15 minutes before being heated at 80 °C for 1 hour. The mixture was allowed to cool to room temperature before a mixture of 5N hydrogen chloride (15 ml_, 75 mmol) and 1 ,4-dioxane (30 ml_) was added. The mixture was heated at 80 °C with vigorous stirring for 4 hours. The mixture was allowed to cool to room temperature and was extracted with ethyl acetate. The ethyl acetate layer was back- extracted 1 time with 1 N hydrochloric acid. The organic layer was discarded and the combined acidic aqueous layers were basified with 2M sodium hydroxide and extracted 3 times with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give 2-(4-methoxy-3-(3- methoxypropoxy)benzyl)spiro[3.3]heptan-2-amine (545 mg, 1 .71 mmol, 89 % yield) as a clear oil. LCMS (ESI) m/z: 320.3 (M+1)+.
Figure imgf000325_0002
[001583] Acetic formic anhydride (6.79M in formic acid) (1 .26 ml_, 8.53 mmol) was added dropwise to a 0 °C solution of triethylamine (0.713 ml_, 5.12 mmol) and 2-(4- methoxy-3-(3-methoxypropoxy)benzyl)spiro[3.3]heptan-2-amine (545 mg, 1 .71 mmol) in dichloromethane (DCM) (15 ml_). The mixture was stirred at 0 °C for 2 hours. The mixture was diluted with dichloromethane, washed with saturated sodium bicarbonate, washed with brine, dried over sodium sulfate, and concentrated to give N-(2-(4-methoxy-3-(3- methoxypropoxy)benzyl)spiro[3.3]heptan-2-yl)formamide (593 mg, 1 .71 mmol, 100 % yield).
LCMS (ESI) m/z: 348.3 (M+1)+.
Figure imgf000326_0001
[001585] N-(2-(4-Methoxy-3-(3-methoxypropoxy)benzyl)spiro[3.3]heptan-2- yl)formamide (593 mg, 1 .71 mmol) in acetonitrile (15 mL) were cooled in an ice bath before phosphorus oxychloride (0.191 mL, 2.05 mmol) was added dropwise with stirring. The mixture was heated at 60 °C for 4 hours and concentrated. The residue was dissolved in ethyl acetate and cooled in an ice bath before water and 30% NH4OH were added to bring the pH = ~1 1 . The organic layer was collected and the aqueous layer extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated to give 7"-methoxy-6"-(3-methoxypropoxy)-4"H-dispiro[cyclobutane-1 ,1 '- cyclobutane-3',3"-isoquinoline] (562 mg, 1 .71 mmol, 100 % yield). LCMS (ESI) m/z: 330.3 (M+1)+.
[001586] Step 6: Ethyl 10"-methoxy-9"-(3-methoxypropoxy)-2"-oxo-2".7"- dihydrodispirofcyclobut -alisoquinolinel-3"-carboxylate
Figure imgf000326_0002
[001587] 7"-Methoxy-6"-(3-methoxypropoxy)-4"H-dispiro[cyclobutane-1 ,1 '- cyclobutane-3',3"-isoquinoline] (562 mg, 1 .71 mmol) and ethyl 2-(ethoxymethylene)-3- oxobutanoate (1 .48 mL, 8.53 mmol) in ethanol (6 mL) were heated at reflux overnight. The mixture was cooled, concentrated, and the residue dissolved in 1 ,2-dimethoxyethane (DME) (6 mL). p-Chloranil (419 mg, 1 .71 mmol) was added and the mixture heated at 85 °C for 2 hours. The mixture was cooled in an ice bath. The precipitate was collected by filtration, washed with cold DME, and dried to give ethyl 10"-methoxy-9"-(3-methoxypropoxy)-2"-oxo- 2",7"-dihydrodispiro[cyclobutane-1 ,1 '-cyclobutane-3',6"-pyrido[2,1 -a]isoquinoline]-3"- carboxylate (54 mg, 0.1 15 mmol, 7 % yield) as an off-white solid. LCMS (ESI) m/z: 468.4 (M+1)+. The filtrate was concentrated and the residue purified by silica chromatography eluting with a gradient of 0% to 100% ethyl acetate in dichloromethane. The column was further eluted with a gradient of 0% to 25% ethanol in ethyl acetate. The product eluted at ~15% ethanol. Fractions were concentrated to give ethyl 10"-methoxy-9"-(3- methoxypropoxy)-2"-oxo-2",7"-dihydrodispiro[cyclobutane-1 ,1 '-cyclobutane-3',6"-pyrido[2,1 - a]isoquinoline]-3"-carboxylate (240 mg, 0.513 mmol, 30 % yield) as an oil. LCMS (ESI) m/z:
468.4 (M+1)+. 37% overall yield.
[001588] Step 7: 10"-Methoxy-9"-(3-methoxypropoxy)-2"-oxo-2".7"- dihvdrodispiro[cvclobutane-1 ,1 '-cyclobutane-3', 6"-pyrido[2,1 -alisoquinolinel-3"-carboxylic acid
Figure imgf000327_0001
[001589] A solution of lithium hydroxide monohydrate (264 mg, 6.29 mmol) in water (1 .67 mL) was added to a solution of ethyl 10"-methoxy-9"-(3-methoxypropoxy)-2"- oxo-2", 7"-dihyd rod ispiro[cyclobutane-1 ,1 '-cyclobutane-3', 6"-pyrido[2,1 -a]isoquinoline]-3"- carboxylate (294 mg, 0.629 mmol) in methanol (5 mL) and the mixture heated at 60 °C for 2 hours. The mixture was allowed to cool to room temperature, diluted with water, acidified with 1 M hydrochloric acid, and extracted 3 times with dichloromethane. The combined organic layers were washed with brine and concentrated. The residue was purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient). Fractions were concentrated. The residue was azeotroped with acetonitrile to help remove water. The residue was dissolved in dichloromethane, hexanes added, and the mixture concentrated to give 10"-methoxy-9"-(3-methoxypropoxy)- 2"-oxo-2",7"-dihydrodispiro[cyclobutane-1 ,1 '-cyclobutane-3', 6"-pyrido[2,1 -a]isoquinoline]-3"- carboxylic acid (247 mg, 0.561 mmol, 89 % yield) as a tan solid. 1H NMR (400 MHz, DMSO-c/e) δ ppm 1 .71 - 1 .84 (m, 2 H) 1 .91 - 2.06 (m, 4 H) 2.1 1 - 2.24 (m, 4 H) 2.57 (d, J=13.28 Hz, 2 H) 3.1 1 (s, 2 H) 3.22 (s, 3 H) 3.45 (t, J=6.25 Hz, 2 H) 3.85 (s, 3 H) 4.08 (t, J=6.44 Hz, 2 H) 7.06 (s, 1 H) 7.38 (s, 1 H) 7.45 (s, 1 H) 8.68 (s, 1 H). 28 of 29 protons found. LCMS (ESI) m/z: 440.3 (M+1 )+.
[001590] Example 138
[001591] 1 '.10'-Dichloro-9'-(3-methoxypropoxy)-3.3-dimethyl-2'-oxo-2'.7'- dihydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000327_0002
Figure imgf000328_0001
[001593] Ethyl 10'-chloro-9'-(3-methoxypropoxy)-3,3-dimethyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (13 mg, 0.028 mmol) and NCS (4.53 mg, 0.034 mmol) in acetonitrile (0.5 ml_) were stirred at room temperature for 2 hours and then at 70 °C for 2 hours. The mixture was allowed to cool to room temperature and purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient). Fractions were lyophilized to give ethyl 1 ',10'-dichloro-9'-(3-methoxypropoxy)-3,3-dimethyl-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (10 mg, 0.020 mmol, 72 % yield). 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 8.75 (s, 1 H), 8.38 (s, 1 H), 6.83 (s, 1 H), 4.40 (q, J=7.16 Hz, 2 H), 4.21 (t, J=6.05 Hz, 2 H), 3.62 (t, J=5.86 Hz, 2 H), 3.37 (s, 3 H), 3.1 1 (s, 2 H), 2.37 (d, J=12.49 Hz, 2 H), 2.14 (quin, J=6.05 Hz, 2 H), 1.96 - 2.08 (m, 2 H), 1 .41 (t, J=7.03 Hz, 3 H), 1 .25 (s, 6 H). 29 of 29 protons found. LCMS (ESI) m/z: 494.3 (M+1)+.
[001594] Step 2: 1 ',10'-Dichloro-9'-(3-methoxypropoxy)-3,3-dimethyl-2'-oxo- 2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000328_0002
[001595] A solution of lithium hydroxide monohydrate (7.04 mg, 0.168 mmol) in water (1 ml_) was added to a solution of ethyl 1 ',10'-dichloro-9'-(3-methoxypropoxy)-3,3- dimethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylate (10 mg, 0.017 mmol) in methanol (1 ml_) and the mixture heated at 60 °C for 3 hours. The mixture was allowed to cool to room temperature and purified by medium pressure reverse phase chromatography (C18 / acetonitrile / water / 0.1 % formic acid / 10% to 100% gradient). Fractions were concentrated to give 1 ',10'-dichloro-9'-(3-methoxypropoxy)-3,3- dimethyl-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1-a]isoquinoline]-3'-carboxylic acid (5 mg, 10.6 μηιοΙ, 63.2 % yield). LCMS (ESI) m/z: 466.3 (M+1)+. Ή NMR (400 MHz,
DMSO-c/e) δ ppm 8.85 (s, 1 H), 8.27 (s, 1 H), 7.41 (s, 1 H), 4.23 (t, J=6.25 Hz, 2 H), 3.50 (t, J=6.05 Hz, 2 H), 3.24 (s, 3 H), 1 .88 - 2.08 (m, 4 H), 1 .20 (s, 3 H), 1 .15 (s, 3 H). 20 of 25 protons found. 4 protons under the water and DMSO peaks.
[001596] Example 139
[001597] l O-Cvclopropyl-g-CcvclopropylmethoxyVe-isopropyl-e-methyl^-oxo- 6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000329_0001
[001598] Step 1 : Ethyl 10-cvclopropyl-9-(cvclopropylmethoxy)-6-isopropyl-6- methyl-2-oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000329_0002
[001599] To a solution of ethyl 10-cyclopropyl-9-hydroxy-6-isopropyl-6- methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (38.4 mg, 0.1 mmol) in DMF (2 mL) was added K2C03 (27.4 mg, 0.2 mmol) and (bromomethyl)cyclopropane (20.5 mg, 0.15 mmol). After stirred at 80 °C overnight, the reaction mixture was cooled to room temperature, quenched with H20 and extracted with EtOAc three times. The combined organic layers were washed with brine and dried over Na2S04. The solvent was removed under reduced pressure and subjected to silica gel column purification to afford the title compound (20 mg, 46% yield). LCMS (ESI) m/z: 436.2 (M + 1)+.
[001600] Step 2: 10-Cvclopropyl-9-(cvclopropylmethoxy)-6-isopropyl-6-methyl- 2-0X0-6, 7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000329_0003
[001601] To a solution of ethyl 10-cyclopropyl-9-(cyclopropylmethoxy)-6- isopropyl-6-methyl-2-oxo-6,7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (20 mg, 0.05 mmol) in EtOH (2 mL), and water (1 mL) was added LiOH (6 mg, 0.25 mmol). The mixture was stirred at room temperature The reaction mixture was acidified to pH~1 and was purified by reverse phase reverse phase HPLC (C18 0-40% MeCN in H20 with 0.1 % formic acid) to afford the title compound (7.5 mg, 37% yield) as a white solid. LCMS (ESI) m/: 408.4 (M + 1)+. Ή NMR (400 MHz, DMSO-d6) δ 8.52 (s, 1 H), 7.43 (s, 1 H), 7.38 (s, 1 H), 6.99 (s, 1 H), 4.00 - 3.93 (m, 2H), 3.22 - 3.14 (m, 2H), 2.18 - 2.12 (m,1 H), 1 .80 - 1 .74 (m, 6.8 Hz, 1 H), 1 .65 (s, 3H), 1 .34 - 1 .21 (m, 2H), 0.93 - 0.89 (m, 2H), 0.87 - 0.84 (m, 1 H), 0.80 (dd, J = 6.8 Hz, 3H), 0.61 (dd, J = 6.6 Hz, 5H), 0.41- 0.35 (m, 2H).
[001602] Example 140
[001603] 6-lsopropyl-10-methoxy-9-(3-methoxypropoxy)-7,7-dimethyl-2-oxo-
Figure imgf000330_0001
[001604] Step 1 : 4-Methoxy-3-(3-methoxypropoxy)benzaldehvde
Figure imgf000330_0002
[001605] A mixture of 3-hydroxy-4-methoxybenzaldehyde (1 1 .5 g, 75.6 mmol), 1 -bromo-3-methoxypropane (17.3 g, 1 13.4 mmol) and K2C03 (26.1 g, 189.0 mmol) in MeCN (150 ml) was heated at reflux overnight. The reaction mixture was cooled to room temperature, filtered through a pad of celite, washed with ethyl acatate (50ml x 3) and the filtrate was concentrated under reduced pressure to afford the title compound (16.0 g, 71 .3 mmol, 94 % yield) as a white solid, and was used in the next step directly. LCMS (ESI) m/z: 225.3 (M + 1)+.
[001606] Step 2: -Methoxy-3-(3-methoxypropoxy)phenyl)methanol
Figure imgf000330_0003
[001607] A solution of 4-methoxy-3-(3-methoxypropoxy)benzaldehyde (16.0 g, 71 .3 mmol) in MeOH (150 ml) was cooled to 0°C, NaBH4 (2.71 g, 71 .3 mol) was added portion wise during a period of 30 min. After addition, this mixture was stirred at room temperature for 1 hr, quenched by addition of H20 (30 ml). The organic layer was concentrated under reduced pressure, the residue was dissolved in dichloromethane (180 ml), acidified with 2 N HCI to PH= 6. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to afford 15 g of the title compound as a white solid, which was used as is in the next step. LCMS (ESI) m/z: 227.1 (M + 1)+. [001608] Step 3: 4-(Bromomethyl)-1 -methoxy-2-(3-methoxypropoxy)benzene
Figure imgf000331_0001
[001609] TMSBr (12.6 mL, 99.5 mmol) was added dropwise to a 0°C cooled solution of (4-methoxy-3-(3-methoxypropoxy)phenyl)methanol (15.0 g, 66.3 mmol) in CHCI3 (80 mL). The mixture was stirred at rt for 30 minutes and stood for 1 h. The upper liquid was collected and concentrated. The residue was purified via silica gel chromatography (5-30% EtOAc/PE) to give 19 g of the title compound as a white solid. 1H NMR (400 MHz, CDCI3) δ 6.87 (m, 2H), 6.75 (d, J = 8.0 Hz, 1 H), 4.42 (s, 2H), 4.06 (t, J = 6.5 Hz, 2H), 3.79 (s, 3H), 4.06 (t, J = 6.1 Hz, 2H), 3.29 (s, 3H), 4.06 (p, J = 6.3 Hz, 2H).
[001610] Step 4: -(4-Methoxy-3-(3-methoxypropoxy)phenyl)acetonitrile
Figure imgf000331_0002
[001611] To a solution of 4-(bromomethyl)-1 -methoxy-2-(3- methoxypropoxy)benzene (8.0 g, 27.6 mmol) in DCM (80 mL) was added TMSCN (5.5 g, 55.2 mmol) in one portion and then TBAF (14.4 g, 55.2 mmol) portion wise at 0°C over 10min. The reaction mixture was stirred at rt overnight. The reaction mixture was absorbed on silica gel. (100-200 mesh) and was purified via silica gel chromatography (0-10%
EtOAc/PE) to give the title compound (5.0 g, 21 .25 mmol, 77.0 % yield) as a colorless liquid. LCMS (ESI) m/z: 236.4 (M+H)+.
[001612] Step 5: 2-(4-Methoxy-3-(3-methoxypropoxy)phenyl)-2- methylpropanenitrile
Figure imgf000331_0003
[001613] To a suspension of NaH (1 .17 g, 31 .87 mmol) in THF (80 mL) was added 2-(4-methoxy-3-(3-methoxypropoxy)phenyl)acetonitrile (3.0 g, 12.75 mmol) dropwise at 0°C under N2. The reaction mixture was then stirred for 1 hr at room temperature, followed by the addition of iodomethane (4.5 g, 31 .87 mmol) over 30 min. The resulting mixture was stirred at room temperature overnight. The mixture was quenched with saturated aqueous NH4CI carefully at 0°C and extracted with ethyl acetate. The organic layer was evaporated and the residue was purified by silica gel with ethyl acetate in petroleum ether (0-10 % v/v) to give the title compound (3.0 g, 1 1 .39 mmol, 89.3.0 % yield) as a colorless liquid. LCMS (ESI) m/z: 264.5. (M + 1 )+. [001614] Step 6: 2-(4-Methoxy-3-(3-methoxypropoxy)phenyl)-2,4- dimethylpentan-3-imine
Figure imgf000332_0001
[001615] To a solution of 2-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2- methylpropanenitrile (2.0 g, 7.59 mmol) in THF (30 mL) was added dropwise a solution of Lithium isopropyl (1 .0 M in Hexane, 8.35 mL, 8.35 mmol) at -50°C. Upon completion of addition, the reaction mixture was stirred at -50°C for 30min and -20 °C for another 1 hr. The mixture was quenched with a saturated aqueous NH4CI carefully at 0°C and extracted with ethyl acetate. The organic layer was evaporated and the residue was purified by silica gel with ethyl acetate in petroleum ether (0-50 % v/v) to give the title compound (1 .1 g, 3.58 mmol, 47.1 % yield) as a white solid. LCMS (ESI) m/z: 308.5. (M + 1)+.
[001616] Step 7: 2-(4-Methoxy-3-(3-methoxypropoxy)phenyl)-2,4- dimethylpentan-3-amine
Figure imgf000332_0002
[001617] A solution of 2-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,4- dimethylpentan-3-imine (1 .1 g, 3.58 mmol) in MeOH (30 ml) was cooled to 0°C. NaBH4 (272 mg, 7.2 mol) was added in three portions during a period of 2 min. After addition, this mixture was stirred at room temperature for 1 hr and was quenched by addition of water (5 ml). The reaction mixture was concentrated under reduced pressure, the residue was dissolved in dichloromethane (50 ml) and was acidified with 2 N HCI to pH = 6. The organic layer was washed with brine, dried over Na2S04, filtered and concentrated to afford 1 .1 g of the title compound as a colorless oil, which was used as is in the next step. LCMS (ESI) m/z: 310.5 (M + 1)+.
[001618] Step 8: N-(2-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,4- dimethylpentan-3-yl) formamid
Figure imgf000332_0003
[001619] Acetic formic anhydride (3.18 g, 35.8 mmol) was added dropwise to a solution of 2-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,4-dimethylpentan-3-amine (1 .1 g, 3.58 mmol) and TEA (1 .1 g, 10.7 mmol) in THF (30.0 mL) at 0°C. The mixture was stirred at room temperature for 3 h. The mixture was poured into cool water (50 mL), made basic with 30 % aq. solution of ammonia and extracted with EtOAc (3 x 30 mL). The combined organic extracts were dried over Na2S04 and concentrated under reduced pressure. The residue was purified via silica gel chromatography (0-15% MeOH/DCM) to give the title compound (840 mg, 2.49 mmol, 69.5 % yield) as colorless oil. LCMS (ESI) m/z: 338.5 (M +
[001620] Step 9: 3-lsopropyl-7-methoxy-6-(3-methoxypropoxy)-4,4-dimethyl- 3,4- dihydroisoquinoline
Figure imgf000333_0001
[001621] N-(2-(4-methoxy-3-(3-methoxypropoxy)phenyl)-2,4-dimethylpentan- 3-yl) formamide (840 mg, 2.49 mmol) was dissolved in acetonitrile (20 mL) and was heated with POC (1 .5 mL, 15.9 mmol) at 90 °C for one hour and was concentrated under reduced pressure to give crude, which was dissolved in ethyl acatate (50 ml) and basified with NH4OH until pH>1 1 . The organic layer was separated and was dried over Na2S04. The solvent was evaporated to give 795 mg of the title compound as a gray solid, which was used as is in the next step. LCMS (ESI) m/z: 320.5 (M + 1)+.
[001622] Step 10: Ethyl 6-isopropyl-10-methoxy-9-(3-methoxypropoxy)-7,7- dimethyl-2- oxo-1 ,6,7,1 1 b-tetrahvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000333_0002
[001623] The crude 3-isopropyl-7-methoxy-6-(3-methoxypropoxy)-4,4- dimethyl-3,4- dihydroisoquinoline (795.4 mg, 2.49 mmol) was dissolved in ethanol (30.0 mL) and was treated with ethyl (Z)-2-(ethoxymethylene)-3-oxobutanoate (2.3 g, 12.5 mmol) at 84 °C overnight and was then stirred for two days at 100 °C to drive the reaction to completion. The reaction mixture was concentrated, and was diluted with water and EtOAc. The organic layer was separated, dried over Na2S04, concentrated and the residue was purified via silica gel chromatography (5-100% EtOAc/PE) to give the title compound (289 mg, 0.63 mmol, 25.2 % yield) as a brown oil. LCMS (ESI) m/z: 460.6 (M + 1)+.
[001624] Step 1 1 : Ethyl 6-isopropyl-10-methoxy-9-(3-methoxypropoxy)-7,7- dimethyl-2- oxo-6,7-dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylate
Figure imgf000334_0001
[001625] Ethyl 6-isopropyl-l 0-methoxy-9-(3-methoxypropoxy)-7,7-dimethyl-2- oxo-1 ,6,7,1 1 b-tetrahydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (289 mg, 0.63 mmol) was dissolved in DME (20 mL) and was treated with 2,3,5,6-tetrachlorocyclohexa-2,5-diene- 1 ,4-dione (124 mg, 0.50 mmol) for 16 hr at 80 °C The reaction mixture was concentrated to dryness and the residue was purified via silica gel chromatography (0-5% MeOH/DCM) to give the title compound (1 10 mg, 0.24 mmol, 38.1 % yield) as a black oil. LCMS (ESI) m/z: 458.6 (M + 1)+.
[001626] Step 12: 6-lsopropyl-10-methoxy-9-(3-methoxypropoxy)-7,7- dimethyl-2-oxo-6,7- dihvdro-2H-pyrido[2,1 -alisoquinoline-3-carboxylic acid
Figure imgf000334_0002
[001627] Ethyl 6-isopropyl-10-methoxy-9-(3-methoxypropoxy)-7,7-dimethyl-2- oxo-6, 7-dihydro-2H-pyrido[2,1 -a]isoquinoline-3-carboxylate (1 10 mg, 0.24 mmol) was dissolved in THF (30 ml), Water (5.0 ml) and was treated with LiOH (60 mg, 2.5 mmol) at rt for 1 hr. The reaction mixture was acidified with 1 N HCI to pH=4, and the mixture was concentrated. Water and EtOAc were added and the organic layer was separated. The solvent was concentrated, and the crude product was purified by reverse phase reverse phase HPLC (C18, 0~90 acetonitrile in H20 with 0.1 % formic acid) to provide 6-isopropyl- 10-methoxy-9-(3-methoxypropoxy)-7,7-dimethyl-2-oxo-6,7- dihydro-2H-pyrido [2,1 - a]isoquinoline-3-carboxylic acid (63 mg, 0.15 mmol, 61 .1 % yield) as a white solid. LCMS (ESI) m/z: 430.2 (M + 1)+. 1 H NMR (400 MHz, DMSO-d6) δ 8.70 (s, 1 H), 7.48 (s, 1 H), 7.47 (s, 1 H), 6.99 (s, 1 H), 4.38 (d, J = 3.9 Hz, 1 H), 4.19-4.1 1 (m, 2H), 3.88 (s, 2H), 3.48 (t, J = 6.2 Hz, 2H), 3.26 (s, 3H), 2.09 - 2.01 (m, 1 H), 1 .97 (dd, J = 12.6, 6.3 Hz, 2H), 1 .51 (s, 3H), 1 .08 (s, 3H), 0.94 (d, J = 6.8 Hz, 3H), 0.21 (d, J = 6.9 Hz, 3H).
[001628] Example 141
[001629] Compound 141 may be prepared as shown in Scheme 4, and as detailed below: 9',10'-dimethoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane-1 ,6'-pyrido[2,1 - alisoquinolinel-3'-carboxylic acid
Figure imgf000335_0001
[001630] Step 1 : 4-(chloromethvD-1 ,2-dimethoxybenzene
Figure imgf000335_0002
[001631] To a 0 °C cooled solution of (3,4-dimethoxyphenyl)methanol (10 g,
59.5 mmol) and pyridine (4.81 mL, 59.5 mmol) in dichloromethane (300 mL) was added drop wise thionyl chloride (4.34 mL, 59.5 mmol) and the reaction mixture was stirred at 0 °C for 2 hours. Water was added and the organic layer was separated and then dried over MgS04. The solvent was evaporated and the crude material was purified via silica gel chromatography to give 4-(chloromethyl)-1 ,2-dimethoxybenzene (2.66 g, 14.27 mmol, 24 % yield). Ή NMR (400 MHz, CHLOROFORM-d) δ ppm 6.91 - 6.99 (m, 2 H), 6.84 (d, J=8.1 Hz, 1 H), 4.58 (s, 2 H), 3.90 (d, J=7.3 Hz, 6 H).
Figure imgf000335_0003
[001632] Step 2: ethyl 1 -(3,4-dimethoxybenzyl)cvclobutane-1 -carboxylate
[001633] LDA (1 .0M in hexanes) (19.29 mL, 19.29 mmol) was added drop wise to a -78 °C cooled solution of ethyl cyclobutanecarboxylate (2.472 g, 19.29 mmol) in Tetrahydrofuran (THF) (100 mL) and the reaction mixture was continued with stirring at -78 °C for one hour. Next, 4-(chloromethyl)-1 ,2-dimethoxybenzene (in 10mL THF) (2.4 g, 12.86 mmol) was added, and the resulting reaction mixture was stirred at -78 °C for another hour. The cooling bath was removed and the mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with water and EtOAc, transferred to separatory funnel and the organic layer was separated. The organics were dried over MgS04, concentrated and was purified via silica gel chromatography to give ethyl 1 -(3,4- dimethoxybenzyl)cyclobutane-1 -carboxylate (687 mg, 2.468 mmol, 19.19 % yield). 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 6.74 - 6.81 (m, 1 H), 6.62 - 6.72 (m, 2 H), 4.05 - 4.21 (m, 2 H), 3.76 - 3.91 (m, 6 H), 3.04 (s, 2 H), 2.31 - 2.50 (m, 2 H), 2.00 - 2.13 (m, 2 H), 1 .79 - 1 .96 (m, 2 H), 1 .23 (t, J=7.1 Hz, 3 H), LCMS(ES+)(m/z):279.35 (M+1 ).
[001634] Step 3: 1 -(3,4-dimethoxybenzyl)cvclobutane-1 -carboxylic acid
Figure imgf000336_0001
[001635] To a solution of ethyl 1 -(3,4-dimethoxybenzyl)cyclobutane-1 - carboxylate (659 mg, 2.368 mmol) dissolved in methanol (10 mL), tetrahydrofuran (THF) (5 mL) and water (2.500 mL) was added lithium hydroxide (397 mg, 9.47 mmol). The reaction mixture was stirred overnight at 55 °C, then cooled to 0 °C and acidified to pH 2 with 1 N HCI. The mixture was concentrated, diluted with water and EtOAc, and the organic layer was separated. The organic layer was washed with brine, dried over MgS04 and concentrated. The crude material was purified via silica gel chromatography to give 1 -(3,4- dimethoxybenzyl)cyclobutane-1 -carboxylic acid (582 mg, 2.325 mmol, 98 % yield). 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 6.76 - 6.81 (m, 1 H), 6.70 - 6.76 (m, 2 H), 3.86 (s, 3 H), 3.83 (s, 3 H), 3.07 (s, 2 H), 2.41 - 2.51 (m, 2 H), 2.10 (m, J=1 1 .9, 9.2, 6.0 Hz, 2 H), 1 .88 - 1 .98 (m, 2 H), LCMS(ES+)(m/z):251 .31 (M+1).
[001636]
Figure imgf000336_0002
[001637] A solution of 1 -(3,4-dimethoxybenzyl)cyclobutane-1 -carboxylic acid (557 mg, 2.225 mmol), DPPA (575 μΙ, 2.67 mmol) and Et3N (372 μΙ, 2.67 mmol) in tert- butanol (8366 μΙ) in a sealed tube was purged with nitrogen, and stirred with heating at 90 °C for 4 days. The reaction mixture was cooled to room temperature, EtOAc and water were added and the organic layer was separated. The organic layer was dried over MgS04, then concentrated and purified via silica gel chromatography to give tert-butyl (1 -(3,4- dimethoxybenzyl)cyclobutyl)carbamate (556 mg, 1 .730 mmol, 78 % yield). 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 6.81 (d, J=8.0 Hz, 1 H), 6.56 - 6.75 (m, 2 H), 3.79 - 3.94 (m, 6 H), 3.03 (s, 2 H), 2.01 - 2.21 (m, 4 H), 1 .78 - 1 .99 (m, 2 H), 1 .47 (s, 9 H)
LCMS(ES+)(m/z):322.39 (M+1).
[001638] Step 5: 1 -(3,4-dimethoxybenzyl)cvclobutan-1 -amine hydrochloride
Figure imgf000336_0003
[001639] A solution of tert-butyl (1 -(3,4-dimethoxybenzyl)cyclobutyl)carbamate (527 mg, 1 .640 mmol) and HCI (4.0M in dioxane) (20 mL, 80 mmol) in dioxane (5 mL) was stirred overnight at room temperature. The solvent was evaporated to give 420 mg of crude 1 -(3,4-dimethoxybenzyl)cyclobutan-1 -amine hydrochloride as a white residue, which was used as is in the next step without purification. LCMS(ES+)(m/z):222.30 (M+1).
[001640] Step 6: ybenzyl)cvclobutyl)formamide
Figure imgf000337_0001
[001641] 1 -(3,4-dimethoxybenzyl)cyclobutan-1 -amine hydrochloride (130 mg, 0.504 mmol) was dissolved in dioxane (5.0 ml_) and was treated with a pre-mixed solution consisting of formic acid (5 ml_, 130 mmol) and acetic anhydride (5 ml_, 53.0 mmol) dissolved in dioxane (5.0 ml_), and the mixture was heated at 90 °C with stirring for one hour. The reaction mixture was treated another 3 times with the pre-mixed acetic anhydride (5 ml_, 53.0 mmol) and formic acid (5 ml_, 130 mmol) mixture, with stirring at 90 °C. The reaction mixture was cooled to room temperature and the solvent was evaporated. Water and EtOAc were added and the organic layer was separated, and was dried over MgS04. The solvent was evaporated to give 95 mg of crude N-(1 -(3,4- dimethoxybenzyl)cyclobutyl)formamide as a tan oil, which was used directly in the next step without purification. LCMS(ES+)(m/z):250.32 (M+1 ).
[001642] Step 7: 6',7'-dimethoxy-4'H-spiro[cvclobutane-1 ,3'-isoquinolinel
Figure imgf000337_0002
[001643] N-(1 -(3,4-dimethoxybenzyl)cyclobutyl)formamide (88 mg, 0.353 mmol) was dissolved in acetonitrile (5.00 ml_) and was heated with POCI3 (0.033 ml_, 0.353 mmol) at 90 °C for one hour. The reaction mixture was cooled to 0 °C and was basified with NH4OH until pH>1 1 . EtOAc and water were added, and the organic layer was separated and was dried over MgS04. The solvent was evaporated to give 77 mg of crude 6', 7'- dimethoxy-4'H-spiro[cyclobutane-1 ,3'-isoquinoline] a dark solid, which was used in the next step without purification. LCMS(ES+)(m/z):232.3 (M+1).
[001644] Step 8: ethyl g'.I O'-dimethoxy^'-oxo-l '^'J'.H b1- tetrahydrospirofcyclobutane-1 ,6'-pyrido[2,1 -alisoquinolinel-3'-carboxylate
Figure imgf000337_0003
[001645] The crude 6',7'-dimethoxy-4'H-spiro[cyclobutane-1 ,3'-isoquinoline] was dissolved in ethanol (5.00 ml_) and was treated with ethyl (Z)-2-(ethoxymethylene)-3- oxobutanoate (263 mg, 1 .412 mmol) at 84 °C overnight and was then stirred for three more days at 100 °C to drive the reaction to completion. The reaction mixture was concentrated, and was diluted with water and EtOAc. The organic layer was separated, dried over MgS04 and was concentrated to give 154 mg of crude ethyl 9',10'-dimethoxy-2'-oxo-1 ',2',7',1 1 b'- tetrahydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate, which was used directly in the next step without purification. LCMS(ES+)(m/z):372.4 (M+1 ).
[001646] Step 9: ethyl 9',10'-dimethoxy-2'-oxo-2',7'-dihvdrospiro[cyclobutane- 1 ,6'-pyrido[2,1 -alisoquinolinel- '-carboxylate
Figure imgf000338_0001
[001647] The crude ethyl 9',10'-dimethoxy-2'-oxo-1 ',2',7',1 1 b'- tetrahydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate was dissolved in DME (5 ml_) and was treated with 2,3,5,6-tetrachlorocyclohexa-2,5-diene-1 ,4-dione (87 mg, 0.353 mmol) for three hours at 80 °C. The reaction mixture was concentrated, then diluted with water and EtOAc, and the organic layer was separated. The organic layer was dried over MgS04 and the solvent was evaporated. The resulting residue was suspended in cold DME and a dark tan solid was filtered off to give 83 mg of crude ethyl 9',10'-dimethoxy-2'- oxo-2', 7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (~90% clean by LCMS) which was used directly in the next step without purification.
LCMS(ES+)(m/z):370.4 (M+1 ).
[001648] Step 10: 9',10'-dimethoxy-2'-oxo-2',7'-dihvdrospiro[cvclobutane-1 ,6'- pyrido[2,1 -alisoquinolinel-3'-carboxylic acid
Figure imgf000338_0002
[001649] To a solution of crude ethyl 9',10'-dimethoxy-2'-oxo-2',7'- dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylate (40 mg, 0.108 mmol) in MeOH (3.00 mL) and Water (3 mL) was added LiOH (38.72 mg, 0.923 mmol) and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was neutralized to pH4 with HCI, and the mixture was diluted with water and EtOAc. The organic layer was separated and was dried over MgS04. The solvent was evaporated and the crude material was purified via reverse phase HPLC (10-90% MeCN/H20, 0.1 %FA) to give 9', 10'- dimethoxy-2'-oxo-2',7'-dihydrospiro[cyclobutane-1 ,6'-pyrido[2,1 -a]isoquinoline]-3'-carboxylic acid (28 mg, 0.078 mmol, 72.0 % yield) as a tan solid. Ή NMR (400 MHz, CHLOROFORM- d) δ ppm 8.97 (s, 1 H), 7.19 (s, 1 H), 7.1 1 (s, 1 H), 6.83 (s, 1 H), 4.02 (s, 3 H), 3.99 (s, 3 H), 3.26 (s, 2 H), 2.61 - 2.74 (m, 2 H), 2.22 (ddt, J=10.4, 7.9, 3.0, 3.0 Hz, 2 H), 2.06 (dd, J=9.2, 6.4 Hz, 2 H). LCMS(ES+)(m/z):342.3 (M+1 ).
[001650] Example 142 - Other Compounds
[001651] Compounds 142-207, 215-222 and 237-276 as disclosed in Table 1 B may be made according to Scheme 1.
[001652] Example 143 - Diastereomers
[001653] Compounds 142-207, 215-222 and 237-278 as disclosed in Table 1 B may also be made as a mixture of diastereomers, as shown in Scheme 2.
[001654] Example 144
[001655] Compounds 208-214 and 223-236 as disclosed in Table 1 B may be prepared as generally shown in Scheme 3.
[001656] Biological Examples
[001657] Example 145 Compound treatment in primary human hepatocvtes infected with HBV
[001658] HBV-Aq inhibitors
[001659] Vials of cryopreserved primary human hepatocytes were placed in a 37°C water bath just until thawed. The cells were pooled, resuspended gently in differentiation medium (Williams medium containing differentiation supplement, GlutaMax- 1™, and penicillin/streptomycin) and counted using a hemacytometer. The cells were pelleted by centrifugation at 1000 X g for 10 min and resuspended to a density of 5.5 X 105 cells/mL in differentiation medium. 100 μΙ_ of cell suspension were plated in each well of collagen-coated 96-well plates. The plates were incubated at 37°C and 5% C02 for 2 days prior to infection.
[001660] HBV stocks were prepared by ultrafiltration of media from
HepG2.2.15 cell cultures. To prepare a working virus stock with a multiplicity of infection (MOI) of 100, HBV stock was added to differentiation medium containing 4% polyethylene glycol to achieve a concentration of 5.5 X 107 HBV DNA copies/mL. The cell media was replaced with 100 μΙ_ of the working virus stock in columns 1 -1 1 and with differentiation medium in column 12. The plates were incubated at 37°C and 5% C02 for approximately 24 hr.
[001661] Compounds were resuspended in DMSO and serially diluted 3-fold in DMSO to make a 10-point dilution series at 200X the final desired concentrations.
Columns 1 1 and 12 contained DMSO. Using a Biomek FX, 2.5 μΙ_ of each compound dilution was stamped into 96-well U-bottom plates, making 3 copies. The compound plates were sealed and stored at -20°C. After equilibration to room temperature, the compound plates were diluted 200 fold with assay medium (differentiation medium plus 1 mM ABT). The media on the cell plates was replaced with 150 μΙ_ of the diluted compounds. The final highest compound concentration was 20 μΜ. The plates were incubated at 37°C and 5% C02. The compound treatments were repeated at days 4 and 9 following the initial treatment. Readouts were done of HBsAg and HBeAg ELISAs at days 9 and 14.
[001662] At Days 9 after the initial treatment, the media was transferred from each cell plate to a U-bottom plate and stored at -80°C. At Day 14 after the initial treatment, the cell plates were washed once with PBS and stored at -80°C.
[001663] Hbs Aa ELISA
[001664] Frozen plates containing the collected media were equilibrated to room temperature in a biosafety cabinet for approximately 30 min. The HBsAg ELISA kit was used according to the manufacturer's directions. Briefly, the ELISA plates and solutions were equilibrated to room temperature for approximately 1 hr and the plates were washed once with 300 1 X wash buffer. 100 1X enzyme conjugate solution, 120 PBS containing 10% FBS, and 30 collected media were placed in each well. The assay plates were sealed and incubated at 37°C for approximately 2 hr. The plates were washed 4 times with 195 mL 1 X wash buffer and dried thoroughly inverted on paper towels. 195 of chromagen/substrate solution were added to each well and incubated at room temperature for approximately 5 min. 100 of stop solution were added to each well and the plates were read on a Molecular Devices SpectraMax 384 Plus Microplate Reader at 450 nm. IC50S were determined using GraphPad Prism: Four-parameter logistic curve with equation Y=Bottom + (Top-Bottom)/(1 +10A((LoglC50-X)*HillSlope))
[001665] HBeAq ELISA
[001666] Frozen plates containing the collected media were equilibrated to room temperature in a biosafety cabinet for approximately 30 min. The HBeAg ELISA kit was used according to the manufacturer's directions. Briefly, the ELISA plates and solutions were equilibrated to room temperature for approximately 1 hr and the plates were washed once with 300 μΐ 1 X wash buffer. 80 μΐ PBS containing 10% FBS, and 20 μΐ collected media were placed in each well. The assay plates were sealed and incubated at 37°C for approximately 1 hr. The plates were washed twice with 300 mL 1X wash buffer and dried thoroughly inverted on paper towels. 100 μί 1X enzyme conjugate solution was placed in each well. The assay plates were sealed and incubated at 37°C for at least 1 hr. The plates were washed 3 times with 300 mL 1X wash buffer and dried thoroughly inverted on paper towels. 100 μί chromagen/substrate solution were added to each well and incubated at room temperature for approximately 5 min. 100 μί of stop solution were added to each well and the plates were read on a Molecular Devices SpectraMax 384 Plus Microplate Reader at 450 nm.
[001667] Data analysis
[001668] IC5oS were determined using GraphPad Prism: Four-parameter logistic curve with equation Y=Bottom + (Top-Bottom)/(1 +10A((LoglC50-X)*HillSlope)). IC50 values for 50% reduction in HBs and HBe antigens for tested compounds are shown in Table 1 for days 9 and 14. As can be seen from the IC50 values, the tested compounds exhibited 50% inhibition of HBe and HBs antigens at values between less than ~0.22 μΜ and 0.0069 μΜ.
[001669] The embodiments of the invention described above are intended to be merely exemplary; numerous variations and modifications will be apparent to those skilled in the art. All such variations and modifications are intended to be within the scope of the present invention as defined in any appended claims.
0
TABLE 1A
Figure imgf000342_0001
0
Figure imgf000343_0001
0
Figure imgf000344_0001
0
Figure imgf000345_0001
0
Figure imgf000346_0001
0
Figure imgf000347_0001
0
Figure imgf000348_0001
0
Figure imgf000349_0001
0
Figure imgf000350_0001
0
Figure imgf000351_0001
0
Figure imgf000352_0001
0
Figure imgf000353_0001
0
Figure imgf000354_0001
0
Figure imgf000355_0001
0
Figure imgf000356_0001
0
Hep AD 38,
Example
Structure Name HBsAg No.
EC50 (uM)
7-cyclohexyl-6-isopropyl-9/10-dimethoxy-2-oxo-6/7-dihydro-2H-
185 pyrido[2,l-a]isoquinoline-3-carboxylic acid -
6-isopropyl-9,10-dimethoxy-7-(oxetan-3-yl)-2-oxo-6/7-dihyd ro-2H-
186 pyrido[2,l-a]isoquinoline-3-carboxylic acid -
6-isopropyl-9,10-dimethoxy-2-oxo-7-(tetrahydrofuran-3-yl)-6/7-
187 dihydro-2H-pyrido[2,l-a]isoquinoline-3-carboxylic acid -
6-isopropyl-9,10-dimethoxy-2-oxo-7-(tetrahydro-2H-pyran-4-yl)-
188 6,7-dihydro-2H-pyrido[2/l-a]isoquinoline-3-carboxylic acid -
6-isopropyl-9,10-dimethoxy-2-oxo-7-(tetrahydrofuran-2-yl)-6/7-
189 dihydro-2H-pyrido[2,l-a]isoquinoline-3-carboxylic acid -
6-isopropyl-9,10-dimethoxy-2-oxo-7-(tetrahydro-2H-pyran-3-yl)-
190 6,7-dihydro-2H-pyrido[2/l-a]isoquinoline-3-carboxylic acid -
6-isopropyl-9,10-dimethoxy-2-oxo-7-(tetrahydro-2H-pyran-2-yl)-
191 6,7-dihydro-2H-pyrido[2/l-a]isoquinoline-3-carboxylic acid -
7-(dimethylamino)-6-isopropyl-9/10-dimethoxy-2-oxo-6/7-dihydro-
192 2H-pyrido[2,l-a]isoquinoline-3-carboxylic acid -
6-isopropyl-9,10-dimethoxy-2-oxo-7-(piperazin-l-yl)-6/7-dihydro-
193 2H-pyrido[2,l-a]isoquinoline-3-carboxylic acid -
6-isopropyl-9,10-dimethoxy-2-oxo-7-phenyl-6/7-dihydro-2H-
194 pyrido[2,l-a]isoquinoline-3-carboxylic acid - 0
Figure imgf000358_0001
0
Figure imgf000359_0001
0
Figure imgf000360_0001
0
Figure imgf000361_0001
0
Figure imgf000362_0001
0
Figure imgf000363_0001
0
Figure imgf000364_0001

Claims

0 What is claimed is:
1 . A compound of Formula I
Figure imgf000365_0001
Formula I wherein
R1 and R4 are independently selected from hydrogen; halogen; hydroxy; alkoxy; amino; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci_ 6alkoxy or halo- or alkyl-substituted Ci-6alkyoxy; unsubstituted C3-7cycloalkyl or halo- or alkyl- substituted C3-7cycloalkyl; unsubstituted C3-7cycloalkenyl or halo- or alkyl-substituted C3. ycycloalkenyl; unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl; and unsubstituted -(Ci-6alkyl)N- CxH2x-phenyl or halo- or alkyl-substituted -(Ci-6alkyl)N- CxH2x- phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
R2 and R3 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR20 R21 ; COR19; COOR19; CONR20 R21 ; pyrrolidinyl; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; Ci-6alkylCOOR19; -BOR19OR19'; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_6alkyoxy; unsubstituted C3-7cycloalkyl or halo- or alkyl- substituted C3.7cycloalkyl; unsubstituted C3.7cycloalkenyl or halo- or alkyl-substituted C3. ycycloalkenyl; unsubstituted or halo- or alkyl-substituted monocyclic heterocycloalkyi ring wherein a heteroatom in the heterocycloalkyi ring is O, N or S; unsubstituted or halo- or alkyl-substituted heteroaryl ring wherein a heteroatom in the heteroaryl ring is O, N or S; pyrrolidinyl; furanyl; pyrrolyl; imidazolyl; unsubstituted -CxH2x-phenyl or halo- or alkyl- substituted -CxH2x-phenyl; unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -O- CxH2x-phenyl; unsubstituted -(Ci-6alkyl)N- CxH2x-phenyl or halo- or alkyl-substituted -(Ci_ 6alkyl)N- CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;and -OR12; or R2 and R3 together form a 3 to 8 membered cycloalkyl ring; or R2 and R3 together form a 3 to 8 membered heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O, N, NR22, S, SR22 or SR22R22'; or R2 and R3 together form a 3 to 8 member aryl ring; or R2 and R3 together 0 form a 3 to 8 member heteroaryl ring, wherein a heteroatom in the heteroaryl ring is O, N, NR22, S, SR22 or SR22R22';
R5 and R6 are independently selected from halogen; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci_6alkoxy or halo- or alkyl-substituted Ci_ 6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3-8cycloalkyl; unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl and heteroaryl; or R5 and R6 together form a 3- to 8-membered cycloalkyl ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R13, R13', R14 and/or R14', or R5 and R6 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R13, R13', R14 and/or R14', wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
R7 and R8 are selected from independently hydrogen; halogen; cyano; unsubstituted Ci-6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3.
ecycloalkyl; unsubstituted CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R7 and R8 together form a 3- to 8-membered cycloalkyl ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18', or R7 and R8 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R17, R17', R18 and/or R18', wherein a heteroatom in the heterocycloalkyi ring is O, N, NR22, S, SR22 or SR22R22';
R9 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_ 6alkoxy; unsubstituted C3.8cycloalkyl or halo- or alkyl-substituted C3.8cycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted or halo- or alkyl substituted - 0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
R10 is selected from COOR19 and CONR20 R21 ;
R11 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_ 6alkyoxy; unsubstituted or halo- or alkyl-substituted C3.7cycloalkyl; unsubstituted CxH2x- phenyl or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
R12 is selected from hydrogen; unsubstituted Ci_6alkyl or Ci_6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy,
heterocycloalkyi, heteroaryl, C3.8cycloalkyl and C2.6alkenyl; Ci-6alkylCi-6alkoxy; Ci_6alkylCi- 0
6alkoxyCi-6alkoxy; Ci-8alkylamino; Ci-8alkylaminocarbonylCi-6alkyl; Ci-6alkylaminosulfonylCi- 6alkyl; Ci_6alkylsulfonylCi-6alkyl; Ci-6alkylcyano; Ci-6alkylC3-8cycloalkyl; Ci-6alkylC3- ecycloalkylcyano; Ci-6alkylphenyl; Ci-6alkylcarbonylpyrrolidinyl; C2-6alkynyl; C2.6alkynylCi- 6hydroxy; Ci-6alkylCi-6alkoxyamino; Ci-6alkylCi-6alkoxyaminoCi-6alkyl; Ci-6alkylCi- 6alkoxyaminodiCi-6alkyl; Ci-6alkylcarboxy; Ci-6alkylaminocarbonylCi-6alkoxy; Ci_
6alkylheteroaryl, wherein the heteroaryl is a monocyclic heteroaryl comprising O and/or ; and Ci-6alkylheterocycloalkyl, wherein the heterocycloalkyi is a monocyclic heterocycloalkyi comprising O and/or N;
R13 , R13', R14 and R14' are independently selected from hydrogen,; halogen; amino; aminoalkyl; cyano; Ci_6alkyl; Ci-6alkoxy; carbonyl; carboxamide; and amide; or R13 and R13' or R14 and R14' together form a 3- to 8-membered cycloalkyi; or R14 and R14' together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
R17 , R17', R18 and R18' are independently selected from hydrogen; halogen; amino; cyano; Ci_6alkyl; Ci-6alkoxy; carbonyl; carboxamide; and amide; or R17 and R18 or R17' and R18' together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3.
ecycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
R19, R19' and R19" are independently selected from hydrogen; Ci_6alkyl; C3.8cycloalkyl; C2.6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci.6alkyltetrazole; Ci_ 6alkylthiazole; Ci-6alkyloxazole; Ci.6alkyldioxazole; and Ci-6alkyloxazolidone; and
R20 and R21 are independently selected from hydrogen; Ci_6alkyl; C3.8cycloalkyl; C2. 6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci-6alkyltetrazole; Ci_
6alkylthiazole; Ci-6alkyloxazole; Ci.6alkyldioxazole; and Ci-6alkyloxazolidone; or R20 and R21 together with the nitrogen to which they are attached form unsubstituted pyrrolidinyl, unsubstituted piperidinyl, or unsubstituted morpholinyl; or R20 and R21 together with the nitrogen to which they are attached form carboxyl-substituted pyrrolidinyl, carboxyl- substituted piperidinyl or carboxyl-substituted morpholinyl;
R22 and R22' are independently selected from hydrogen; oxygen; Ci_6alkyl; Ci_
6haloalkyl; C3.8cycloalkyl; Ci-6alkylC3-8cycloalkyl; C2.6alkenyl; phenyl; Ci-6alkylimidizole; Ci_ 6alkoxy; Ci-6alkyltriazole; Ci-6alkyltetrazole; Ci-6alkylthiazole; Ci-6alkyloxazole; Ci_
ealkyldioxazole; Ci-6alkyloxazolidone; -COR19; -COOR19'; -CSOR19 "; and -CONR20R21 ; and
R23 is selected from hydrogen, Ci_6alkyl; C3.8cycloalkyl; C2.6alkenyl; phenyl; Ci_
6alkylimidizole; Ci.6alkoxy; Ci-6alkyltriazole; Ci.6alkyltetrazole; Ci.6alkylthiazole; Ci_ 0
6alkyloxazole; Ci-6alkyldioxazole; Ci-6alkyloxazolidone; aryl; heteroaryl; and benzyl, or a pharmaceutically acceptable salt thereof.
2. A compound of Formula I
Figure imgf000368_0001
Formula I wherein
R1 and R4 are independently selected from hydrogen; halogen; hydroxy; alkoxy; amino; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci_ 6alkoxy or halo- or alkyl-substituted Ci-6alkyoxy; unsubstituted C3-7cycloalkyl or halo- or alkyl- substituted C3-7cycloalkyl; unsubstituted C3-7cycloalkenyl or halo- or alkyl-substituted C3. ycycloalkenyl; unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl; and unsubstituted -(Ci-6alkyl)N- CxH2x-phenyl or halo- or alkyl-substituted -(Ci-6alkyl)N- CxH2x- phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
R2 and R3 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR20 R21 ; COR19; COOR19; CONR20 R21 ; pyrrolidinyl; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; Ci-6alkylCOOR19; -BOR19OR19'; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci-6alkyoxy; unsubstituted C3.7cycloalkyl or halo- or alkyl- substituted C3.7cycloalkyl; C3.7cycloalkenyl or halo- or alkyl-substituted C3.7cycloalkenyl; unsubstituted or halo- or alkyl-substituted monocyclic heterocycloalkyi ring wherein a heteroatom in the heterocycloalkyi ring is O, N or S; unsubstituted or halo- or alkyl- substituted heteroaryl ring wherein a heteroatom in the heteroaryl ring is O, N or S, pyrrolidinyl, furanyl, pyrrolyl, imidazolyl, substituted -CxH2x-phenyl or unsubstituted -CxH2x- phenyl, substituted -0-CxH2x-phenyl or unsubstituted -0-CxH2x-phenyl, substituted -(Ci_ 6alkyl)N- CxH2x-phenyl or unsubstituted -(Ci-6alkyl)N- CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; and -OR12; or R2 and R3 together form a 3 to 8 membered cycloalkyl ring; or R2 and R3 together form a 3 to 8 membered heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O, N, NR22, S, SR22 or SR22R22'; or R2 and R3 together form a 3 to 8 0 member aryl ring; or R2 and R3 together form a 3 to 8 member heteroaryl ring, wherein a heteroatom in the heteroaryl ring is O, N, NR22, S, SR22 or SR22R22';
R5 is selected from hydrogen, unsubstituted Ci_6alkyl, and halo- or alkyl-substituted Ci-6alkyl;
R6 and R7 together form a 3- to 8-membere cycloalkyl ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R15, R15', R16 and/or R16'; or R6 and R7 together form a 3- to 8- membered heterocycloalkyi ring comprising one heteroatom or two or more heteroatoms, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R15, R15', R16 and/or R16', wherein the one heteroatom in the heterocycloalkyi ring is NR20 and the two or more heteroatoms are selected from N, NR22, O, S, SR22 and SR22R22';
R8 is selected from hydrogen, unsubstituted Ci_6alkyl, and halo- or alkyl-substituted Ci-6alkyl;
R9 is selected from hydrogen, hydroxy, halogen, cyano, unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl, unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_ 6alkoxy, unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3-8cycloalkyl, unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl an unsubstituted -0-CxH2x- phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
R10 is selected from COOR19 and CONR20 R21;
R11 is selected from hydrogen, hydroxy, halogen, cyano, unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl, unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_ 6alkyoxy; unsubstituted C3-7cycloalkyl or halo- or alkyl substituted C3-7cycloalkyl;
unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
R12 is hydrogen; unsubstituted Ci_6alkyl or Ci-6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, heterocycloalkyi, heteroaryl, C3.8cycloalkyl and C2.6alkenyl; Ci-6alkylCi-6alkoxy; Ci-6alkylCi-6alkoxyCi-6alkoxy; Ci_
8alkylamino; Ci-8alkylaminocarbonylCi-6alkyl; Ci-6alkylaminosulfonylCi-6alkyl; Ci_
6alkylsulfonylCi-6alkyl; Ci-6alkylcyano; Ci-6alkylC3-8cycloalkyl; Ci-6alkylC3-8cycloalkylcyano; Ci_ 6alkylphenyl; Ci-6alkylcarbonylpyrrolidinyl; C2.6alkynyl; C2-6alkynylCi-6hydroxy; Ci_6alkylCi- 6alkoxyamino; Ci-6alkylCi-6alkoxyaminoCi-6alkyl; Ci-6alkylCi-6alkoxyaminodiCi-6alkyl; Ci_ 6alkylcarboxy; Ci-6alkylaminocarbonylCi-6alkoxy; Ci-6alkylheteroaryl, wherein the heteroaryl is an O-containing or N-containing monocyclic heteroaryl; or Ci-6alkylheterocycloalkyl, wherein the heterocycloalkyi is monocyclic heterocycloalkyi;
R15, R15', R16 and R16' are independently hydrogen, halogen, amino, cyano, Ci_6alkyl, or Ci-6alkoxy, wherein any of R15, R15', R16 or R16', together with R5 or R8, form a 3- to 8- 0 membered cycloalkyi ring or a 3-8 membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci_ 6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22'; or any of R15, R15', R16 and/or R16' together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
R19, R19' and R19 'are independently hydrogen, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl, phenyl, Ci-6alkylimidizole, Ci-6alkoxy, Ci-6alkyltriazole, Ci-6alkyltetrazole, Ci-6alkylthiazole, Ci_ 6alkyloxazole, Ci-6alkyldioxazole; Ci-6alkyloxazolidone;
R20 and R21 are independently hydrogen, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl, phenyl, Ci-6alkylimidizole, Ci-6alkoxy, Ci-6alkyltriazole,Ci-6alkyltetrazole, Ci-6alkylthiazole, Ci_ 6alkyloxazole, Ci-6alkyldioxazole; Ci-6alkyloxazolidone, -CONR22R22'; or R20 and R21 together with the nitrogen to which they are attached form unsubstituted pyrrolidinyl, unsubstituted piperidinyl, or unsubstituted morpholinyl; or form carboxyl-substituted pyrrolidinyl, carboxyl- substituted piperidinyl or carboxyl-substituted morpholinyl;
R22 and R22' are independently selected from hydrogen, oxygen, Ci_6alkyl, Ci_
6haloalkyl, C3-8cycloalkyl, Ci-6alkylC3-8cycloalkyl, C2-6alkenyl, phenyl, Ci-6alkylimidizole, Ci_ 6alkoxy, Ci-6alkyltriazole,Ci-6alkyltetrazole, Ci-6alkylthiazole, Ci-6alkyloxazole, Ci_
ealkyldioxazole; Ci-6alkyloxazolidone, -COR19, -COOR19', -CSOR19 ", -CONR20R21 ; and
R23 is hydrogen, Ci_6alkyl, C3.8cycloalkyl, C2-6alkenyl, phenyl, Ci-6alkylimidizole, Ci_ 6alkoxy, Ci-6alkyltriazole, Ci.6alkyltetrazole, Ci.6alkylthiazole, Ci.6alkyloxazole, Ci_
6alkyldioxazole; Ci.6alkyloxazolidone; aryl, heteroaryl or benzyl, or a pharmaceutically acceptable salt thereof.
3. A compound of Formula I
Figure imgf000370_0001
Formula I wherein 0
R1, and R4 are independently selected from hydrogen; halogen; hydroxy; alkoxy; amino; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci_ 6alkoxy or halo- or alkyl-substituted Ci-6alkyoxy; unsubstituted C3-7cycloalkyl or halo- or alkyl- substituted C3-7cycloalkyl; unsubstituted C3-7cycloalkenyl or halo- or alkyl-substituted C3. ycycloalkenyl; unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl; and unsubstituted -(Ci-6alkyl)N- CxH2x-phenyl or halo- or alkyl-substituted -(Ci-6alkyl)N- CxH2x- phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
R2 and R3 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR20 R21 ; COR19; COOR19; CONR20 R21 ; pyrrolidinyl; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; Ci-6alkylCOOR19'; -BOR19OR19'; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci-6alkyoxy; C3-7cycloalkyl or halo- or alkyl-substituted C3.
ycycloalkyl; unsubstituted C3-7cycloalkenyl or halo- or alkyl-substituted C3-7cycloalkenyl; unsubstituted or halo- or alkyl-substituted monocyclic heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O, N or S; unsubstituted or halo- or alkyl- substituted heteroaryl ring wherein a heteroatom in the heteroaryl ring is O, N or S;
pyrrolidinyl; furanyl; pyrrolyl; imidazolyl; unsubstituted -CxH2x-phenyl or halo- or alkyl- substituted -CxH2x-phenyl; unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -O- CxH2x-phenyl; unsubstituted -(Ci-6alkyl)N- CxH2x-phenyl or halo- or alkyl-substituted -(Ci_ 6alkyl)N- CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; and -OR12; or R2 and R3 together form a 3 to 8 member cycloalkyl ring; or R2 and R3 together form a 3 to 8 member heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O, N, NR22, S, SR22 or SR22R22'; or R2 and R3 together form a 3 to 8 member aryl ring; or R2 and R3 together form a 3 to 8 member heteroaryl ring, wherein a heteroatom in the heteroaryl ring is O, N, NR22, S, SR22 or SR22R22';
R5 and R6 are independently selected from hydrogen; halogen; cyano; unsubstituted Ci-6alkyl or halo-or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3.8cycloalkyl or halo- or alkyl-substituted C3.
ecycloalkyl; unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R5 and R6 together form a 3- to 8-membered cycloalkyl ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R13, R13', R14 and/or R14', or R5 and R6 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R13, R13', R14 and/or R14', wherein a heteroatom in the heterocycloalkyi ring is O, N, NR22, S, SR22 or SR22R22'; 0
R7 and R8 are independently selected from halogen; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_ 6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3-8cycloalkyl; unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; and heteroaryl; or R7 and R8 together form a 3- to 8-membered cycloalkyl ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18', or R7 and R8 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R17 , R18', R17 and/or R18', wherein a heteroatom in the heterocycloalkyi ring is O, N, NR22, S, SR22 or SR22R22';
R9 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_6alkyl or halo-or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_ 6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3-8cycloalkyl; unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted -0-CxH2x- phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
R10 is selected from COOR19and CONR20 R21 ;
R11 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_ 6alkyoxy; unsubstituted or halo- or alkyl-substituted C3.7cycloalkyl; unsubstituted -CxH2x- phenyl or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6;
R12 is selected from hydrogen; unsubstituted Ci_6alkyl or Ci-6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, C3.8cycloalkyl and C2.6alkenyl; Ci-6alkylCi-6alkoxy; Ci-6alkylCi-6alkoxyCi-6alkoxy; Ci-8alkylamino; Ci_ 8alkylaminocarbonylCi-6alkyl; Ci-6alkylaminosulfonylCi-6alkyl; Ci-6alkylsulfonylCi_6alkyl; Ci_ 6alkylcyano; Ci-6alkylC3-8cycloalkyl; Ci-6alkylC3-8cycloalkylcyano; Ci-6alkylphenyl; Ci_ 6alkylcarbonylpyrrolidinyl; C2.6alkynyl; C2-6alkynylCi-6hydroxy; Ci-6alkylCi-6alkoxyamino; Ci_ 6alkylCi-6alkoxyaminoCi-6alkyl; Ci-6alkylCi-6alkoxyaminodiCi-6alkyl; Ci-6alkylcarboxy; Ci_ 6alkylaminocarbonylCi-6alkoxy; Ci-6alkylheteroaryl, wherein the heteroaryl is a monocyclic heteroaryl comprising O and/or ; and Ci-6alkylheterocycloalkyl, wherein the heterocycloalkyi is a monocyclic heterocycloalkyi comprising O and/or N;
R13 , R13', R14 and R14' are independently selected from hydrogen; halogen; amino; aminoalkyl; cyano; Ci_6alkyl; Ci-6alkoxy; carbonyl; carboxamide; and amide; or R13 and R14 or R13' and R14' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_ 0
6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci.6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
R17, R17', R18 and R18' are independently selected from hydrogen; halogen; amino; cyano; Ci_6alkyl; and Ci_6alkoxy; or R17 and R18 or R17' and R18' together form a 3- to 8- membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci_ 6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
R19, R19' and R19" are independently selected from hydrogen; Ci_6alkyl; C3-8cycloalkyl; C2-6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci.6alkyltetrazole; Ci_ 6alkylthiazole; Ci-6alkyloxazole; Ci.6alkyldioxazole; and Ci-6alkyloxazolidone; and
R20 and R21 are independently selected from hydrogen; Ci_6alkyl; C3-8cycloalkyl; C2. 6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci-6alkyltetrazole; Ci_
6alkylthiazole; Ci-6alkyloxazole; Ci.6alkyldioxazole; and Ci-6alkyloxazolidone; or R20 and R21 together with the nitrogen to which they are attached form unsubstituted pyrrolidinyl, unsubstituted piperidinyl, or unsubstituted morpholinyl; or form carboxyl-substituted pyrrolidinyl, carboxyl-substituted piperidinyl or carboxyl-substituted morpholinyl; and
R22 and R22' are independently selected from hydrogen; oxygen; Ci_6alkyl; Ci_ 6haloalkyl, C3-8cycloalkyl; Ci-6alkylC3-8cycloalkyl; C2.6alkenyl; phenyl; Ci-6alkylimidizole; Ci_ 6alkoxy; Ci-6alkyltriazole; Ci-6alkyltetrazole; Ci-6alkylthiazole; Ci-6alkyloxazole; Ci_
ealkyldioxazole; Ci-6alkyloxazolidone; -COR19; -COOR19'; -CSOR19 "; and -CONR20R21 ; and
R23 is selected from hydrogen; Ci_6alkyl; C3.8cycloalkyl; C2.6alkenyl; phenyl; Ci_ 6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci.6alkyltetrazole; Ci.6alkylthiazole; Ci_
6alkyloxazole; Ci.6alkyldioxazole; Ci-6alkyloxazolidone; aryl; heteroaryl and benzyl, or a pharmaceutically acceptable salt thereof.
4. A compound of Formula IA or Formula IB:
Figure imgf000373_0001
Formula I Formula IA Formula IB wherein C* is a carbon atom stereocenter which has a configuration which is (R) or (S); 0
R1, R2, R3 and R4 are independently selected from hydrogen; hydroxy; alkoxy;
halogen; cyano; NR20 R21 COR19; COOR19; CONR20 R21 ; pyrrolidinyl; unsubstituted Ci_6alkyl or halo- or alkyl substituted Ci_6alkyl; Ci-6alkylCOOR19; -BOR19OR19'; unsubstituted Ci- 6alkoxy or halo-or alkyl-substituted Ci_6alkyoxy; unsubstituted C3-7cycloalkyl or halo-or alkyl- substituted C3-7cycloalkyl; unsubstituted C3-7cycloalkenyl or halo- or alkyl-substituted C3. ycycloalkenyl; unsubstituted or halo- or alkyl-substituted monocyclic heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O, N or S; unsubstituted or halo- or alkyl-substituted heteroaryl ring wherein a heteroatom in the heteroaryl ring is O, N or S; pyrrolidinyl; furanyl; pyrrolyl; imidazolyl; unsubstituted or halo- or alkyl-substituted -CxH2x- phenyl; unsubstituted or halo- or alkyl-substituted -0-CxH2x-phenyl; unsubstituted or halo- or alkyl-substituted -(Ci-6alkyl)N- CxH2x-phenyl wherein x is 0, 1 , 2, 3, 4, 5 or 6; and -OR12; or R2 and R3 together form a 3 to 8 member cycloalkyi ring; or R2 and R3 together form a 3 to 8 member heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O, N, NR22, S, SR22 or SR22R22'; or R2 and R3 together form a 3 to 8 member aryl ring; or R2 and R3 together form a 3 to 8 member heteroaryl ring, wherein a heteroatom in the heteroaryl ring is O, N, NR22, S, SR22 or SR22R22';
and
(A) R5 and R6 are independently selected from halogen; cyano; unsubstituted Ci_ 6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3.
ecycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl; unsubstituted or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; and heteroaryl; or R5 and R6 together form a 3- to 8-membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R13, R13', R14 and/or R14', or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R13, R13', R14 and/or R14', wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; and
R7 and R8 are independently hydrogen; halogen; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_ 6alkoxy; unsubstituted C3.8cycloalkyl or halo- or alkyl-substituted C3.8cycloalkyl; unsubstituted or halo- or alkyl-substituted-CxH2x-phenyl; unsubstituted or halo- or alkyl-substituted -O- CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R7 and R8 together form a 3- to 8- membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18', or R7 and R8 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, 0 cyano, R17, R17', R18 and/or R18', wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; or
(B) R5 and R8 are independently selected from hydrogen, unsubstituted Ci_6alkyl, and halo- or alkyl-substituted Ci_6alkyl; and
R6 and R7 together form a 3- to 8-membered cycloalkyl ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R15, R15', R16 and/or R16'; or R6 and R7 together form a 3- to 8- membered heterocycloalkyi ring comprising one heteroatom or two or more heteroatoms, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R15, R15', R16 and/or R16', wherein the one heteroatom in the heterocycloalkyi ring is NR20 and the two or more heteroatoms are selected from N, NR22, O, S, SR22 and SR22R22'; or
(C) R5 and R6 are independently selected from hydrogen; halogen; cyano;
unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3. ecycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl; unsubstituted or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R5 and R6 together form a 3- to 8-membered cycloalkyl ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18', or R5 and R6 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R13 , R13', R14 and/or R14', wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; and
R7 and R8 are independently selected from halogen; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_ 6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3-8cycloalkyl; unsubstituted or halo- or alkyl-substituted
Figure imgf000375_0001
unsubstituted or halo- or alkyl-substituted -O- CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl; and heteroaryl; or R7 and R8 together form a 3- to 8-membered cycloalkyl ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18', or R7 and R8 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R17 , R18', R17 and/or R18', wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; OF
and wherein
R9 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_ 6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3-8cycloalkyl; unsubstituted 0 or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted or halo- or alkyl-substituted-
Figure imgf000376_0001
wherein x is 0, 1 , 2, 3, 4, 5 or 6;
R10 is selected from COOR19and CONR20 R21 ;
R11 is selected from hydrogen; hydroxy; halogen; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_ 6alkyoxy; unsubstituted or halo- or alkyl-substituted C3-7cycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted or halo- or alkyl-substituted -0-CxH2)r- phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6;
R12 is selected from hydrogen; unsubstituted Ci_6alkyl or Ci-6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy,
heterocycloalkyi, heteroaryl, C3-8cycloalkyl and C2-6alkenyl; Ci-6alkylCi-6alkoxy; Ci-6alkylCi- 6alkoxyCi-6alkoxy; Ci.8alkylamino; Ci-8alkylaminocarbonylCi-6alkyl; Ci-6alkylaminosulfonylCi- 6alkyl; Ci_6alkylsulfonylCi-6alkyl; Ci-6alkylcyano; Ci-6alkylC3-8cycloalkyl; Ci-6alkylC3- ecycloalkylcyano; Ci-6alkylphenyl; Ci-6alkylcarbonylpyrrolidinyl; C2-6alkynyl; C2-6alkynylCi- 6hydroxy; Ci-6alkylCi-6alkoxyamino; Ci-6alkylCi-6alkoxyaminoCi-6alkyl; Ci_6alkylCi- 6alkoxyaminodiCi-6alkyl; Ci-6alkylcarboxy; Ci-6alkylaminocarbonylCi-6alkoxy; Ci_
6alkylheteroaryl, wherein the heteroaryl is an O-containing or N-containing monocyclic heteroaryl; or Ci-6alkylheterocycloalkyl, wherein the heterocycloalkyi is a monocyclic heterocycloalkyi;
R13 , R13', R14 and R14' are independently selected from hydrogen; halogen; amino; aminoalkyl; cyano; Ci_6alkyl; Ci-6alkoxy; carbonyl; carboxamide; and amide; or R13 and R13' or R14 and R14' together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_ 6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci.6alkoxy, wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22';
R15, R15', R16 and R16' are independently hydrogen, halogen, amino, cyano, Ci_6alkyl, or Ci-6alkoxy, wherein any of R15, R15', R16 or R16', together with R5 or R8, form a 3- to 8- membered cycloalkyi ring or a 3-8 membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci_ 6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22'; or any of R15, R15', R16 and/or R16' together form a 3- to 8-membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3.8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
R17 , R17', R18 and R18' are independently selected from hydrogen; halogen; amino; cyano; Ci-6alkyl; and Ci-6alkoxy; or R17 and R18 or R17' and R18' together form a 3- to 8- 0 membered cycloalkyi ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci_ 6alkoxy, wherein a heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22';
R19, R19' and R19" are independently selected from hydrogen; Ci_6alkyl; C3-8cycloalkyl; C2-6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci.6alkyltetrazole; Ci_ 6alkylthiazole; Ci-6alkyloxazole; Ci.6alkyldioxazole; and Ci-6alkyloxazolidone; and
R20 and R21 are independently selected from hydrogen; Ci_6alkyl; C3-8cycloalkyl; C2. 6alkenyl; phenyl; Ci-6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci-6alkyltetrazole; Ci_ 6alkylthiazole; Ci-6alkyloxazole; Ci.6alkyldioxazole; and Ci-6alkyloxazolidone; or R20 and R21 together with the nitrogen to which they are attached form unsubstituted pyrrolidinyl, unsubstituted piperidinyl, or unsubstituted morpholinyl; or form carboxyl-substituted pyrrolidinyl, carboxyl-substituted piperidinyl or carboxyl-substituted morpholinyl; and
R22 and R22' are independently selected from hydrogen; oxygen; Ci_6alkyl; Ci_ 6haloalkyl; C3-8cycloalkyl; Ci-6alkylC3-8cycloalkyl; C2-6alkenyl; phenyl; Ci-6alkylimidizole; Ci_ 6alkoxy; Ci-6alkyltriazole; Ci-6alkyltetrazole; Ci-6alkylthiazole; Ci-6alkyloxazole; Ci_ ealkyldioxazole; Ci-6alkyloxazolidone; -COR19; -COOR19'; -CSOR19 "; and -CONR20R21 ; and
R23 is selected from hydrogen; Ci_6alkyl; C3.8cycloalkyl; C2.6alkenyl; phenyl; Ci_ 6alkylimidizole; Ci-6alkoxy; Ci-6alkyltriazole; Ci-6alkyltetrazole; Ci-6alkylthiazole; Ci_
6alkyloxazole; Ci.6alkyldioxazole; Ci-6alkyloxazolidone; aryl; heteroaryl; and benzyl, or a pharmaceutically acceptable salt thereof.
5. The compound or pharmaceutically acceptable salt according to claim 1 , wherein:
R1, R4, R6, R9 and R11 are independently selected from hydrogen, halogen, hydroxy; alkoxy; amino; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; and phenyl;
R2 and R3 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR20 R21 ; COR19; COOR19; CONR20 R21 ; pyrrolidinyl; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; Ci-6alkylCOOR19; -B(OH)2; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci-6alkyoxy; unsubstituted C3.7cycloalkyl or halo- or alkyl-substituted C3. ycycloalkyl; unsubstituted C3.7cycloalkenyl or halo- or alkyl-substituted C3.7cycloalkenyl; unsubstituted or halo- or alkyl-substituted monocyclic heterocycloalkyi ring wherein a heteroatom in the heterocycloalkyi ring is O or N; unsubstituted or halo- or alkyl-substituted heteroaryl ring wherein a heteroatom in the heteroaryl ring is O or N; pyrrolidinyl; furanyl; pyrrolyl; imidazolyl; unsubstituted or halo- or alkyl-substituted -CxH2)^phenyl; unsubstituted or halo- or alkyl-substituted -0-CxH2)^phenyl; unsubstituted or halo- or alkyl-substituted - 0
(Ci-6alkyl)N- CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; and -OR12; or R2 and R3 together form a 3 to 8 membered cycloalkyl ring; or R2 and R3 together form a 3 to 8 membered heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O or N;
R5 and R6 are independently selected from halogen; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_ 6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3-8cycloalkyl; unsubstituted -CxH2x-phenyl or halo- or alkyl-substituted -CxH2x-phenyl; unsubstituted -0-CxH2x-phenyl or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; aryl and heteroaryl; or R5 and R6 together form a 3- to 8-membered cycloalkyl ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R13, R13', R14 and/or R14', or R5 and R6 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R13, R13', R14 and/or R14', wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
R7 and R8 are independently selected from hydrogen, halogen; cyano; unsubstituted Ci-6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl- substituted Ci-6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3.
ecycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted or halo- or alkyl-substituted -0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R7 and R8 together form a 3- to 8-membered cycloalkyl ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18', or R7 and R8 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R17, R17', R18 and/or R18', wherein a heteroatom in the heterocycloalkyi ring is O or N;
R10 is COOR19; and
R12 is hydrogen; unsubstituted Ci_6alkyl or Ci_6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, heterocycloalkyi, heteroaryl, C3.8cycloalkyl and C2.6alkenyl; Ci-6alkylCi-6alkoxy; Ci-6alkylphenyl; Ci-6alkylcarboxy; Ci_ 6alkylheteroaryl, wherein the heteroaryl is a monocyclic heteroaryl comprising O and/or N; or Ci-6alkylheterocycloalkyl, wherein the heterocycloalkyi is a monocyclic heterocycloalkyi comprising O and/or N; and
R19, R19' and R19" are independently selected from hydrogen; Ci_6alkyl; and phenyl.
6. The compound or pharmaceutically acceptable salt according to claim 5, wherein R4 and R11 are independently selected from H, Br, CI and F; and
R1, R7, R8 and R9 are independently selected from H, unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; and 0
R10 is COOH.
7. The compound or pharmaceutically acceptable salt according to claim 2, wherein: R1 , R4 R5, R8, R9 and R11 are independently selected from hydrogen, unsubstituted Ci_6alkyl, and halo- or alkyl-substituted Ci_6alkyl;
R2 and R3 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR20 R21 ; COR19; COOR19; CONR20 R21 ; pyrrolidinyl; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; Ci-6alkylCOOR19; -B(OH)2; and -OR12;
R6 and R7 together form a 3- to 8-membered cycloalkyl ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R15 , R15', R16 and/or R16'; or R6 and R7 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R15 , R15', R16 and/or R16', wherein a heteroatom in the heterocycloalkyi ring is selected from O, N NR22, S, SR22 and SR22R22';
R12 is hydrogen; unsubstituted Ci_6alkyl or Ci_6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, heterocycloalkyi, heteroaryl, C3-8cycloalkyl and C2-6alkenyl; Ci-6alkylCi-6alkoxy; Ci-6alkylphenyl; Ci-6alkylcarboxy; Ci_ 6alkylheteroaryl, wherein the heteroaryl is a monocyclic heteroaryl comprising O and/or N; or Ci-6alkylheterocycloalkyl, wherein the heterocycloalkyi is a monocyclic heterocycloalkyi comprising O and/or N;
R15, R15', R16 and R16' are independently hydrogen, halogen, amino, cyano, Ci_6alkyl, or Ci-6alkoxy, wherein any of R15, R15', R16 or R16', together with R5 or R8, form a 3- to 8- membered cycloalkyl ring or a 3-8 membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci_ 6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22'; or any of R15, R15', R16 and/or R16' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein a heteroatom in the heteroalkyl ring is O, N, NR22, S, SR22 or SR22R22';
and
R19, R19' and R19" are independently selected from hydrogen; Ci_6alkyl; and phenyl.
8. The compound or pharmaceutically acceptable salt according to claim 7, wherein R1 , R4, R5, R8, R9 and R11 are independently selected from H, Br, CI, F;
R10 is COOH; and
R12 is selected from unsubstituted Ci_6alkyl or Ci_6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, heterocycloalkyi, heteroaryl, 0
C3-8cycloalkyl and C2-6alkenyl.
9. The compound or pharmaceutically acceptable salt according to claim 3, wherein:
R1, R4, R6, R9 and R11 are independently selected from hydrogen, halogen, hydroxy; alkoxy; amino; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; and phenyl;
R2 and R3 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR20 R21 ; COR19; COOR19; CONR20 R21 ; pyrrolidinyl; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; Ci-6alkylCOOR19; -B(OH)2; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci-6alkyoxy; unsubstituted C3-7cycloalkyl or halo- or alkyl-substituted C3- 7cycloalkyl; unsubstituted C3-7cycloalkenyl or halo- or alkyl-substituted C3-7cycloalkenyl; unsubstituted or halo- or alkyl-substituted monocyclic heterocycloalkyi ring wherein a heteroatom in the heterocycloalkyi ring is O or N; unsubstituted or halo- or alkyl-substituted heteroaryl ring wherein a heteroatom in the heteroaryl ring is O or N; pyrrolidinyl; furanyl; pyrrolyl; imidazolyl; unsubstituted or halo- or alkyl-substituted -CxH2)^phenyl; unsubstituted or halo- or alkyl-substituted -0-CxH2x-phenyl; unsubstituted or halo- or alkyl-substituted - (Ci-6alkyl)N- CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; and -OR12; or R2 and R3 together form a 3 to 8 membered cycloalkyi ring; or R2 and R3 together form a 3 to 8 membered heterocycloalkyi ring, wherein a heteroatom in the heterocycloalkyi ring is O or N;
R5 and R6 are independently selected from hydrogen, halogen; cyano; unsubstituted Ci-6alkyl or halo- or alkyl-substituted Ci_6alkyl;
R7 and R8 are independently selected from halogen; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; unsubstituted Ci-6alkoxy or halo- or alkyl-substituted Ci_ 6alkoxy; unsubstituted C3-8cycloalkyl or halo- or alkyl-substituted C3-8cycloalkyl; unsubstituted or halo- or alkyl-substituted -CxH2x-phenyl; and unsubstituted or halo- or alkyl-substituted - 0-CxH2x-phenyl, wherein x is 0, 1 , 2, 3, 4, 5 or 6; or R7 and R8 together form a 3- to 8- membered cycloalkyi ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18'; or R7 and R8 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R17, R17', R18 and/or R18', wherein a heteroatom in the heterocycloalkyi ring is O or N;
R10 is COOH;
R12 is selected from hydrogen; unsubstituted Ci_6alkyl or Ci_6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy,
heterocycloalkyi, heteroaryl, C3-ecycloalkyl and C2-6alkenyl; Ci-6alkylCi-6alkoxy; Ci_
6alkylphenyl; Ci.6alkylcarboxy; Ci-6alkylheteroaryl, wherein the heteroaryl is a monocyclic 0 heteroaryl comprising O and/or N; or Ci-6alkylheterocycloalkyl, wherein the heterocycloalkyi is a monocyclic heterocycloalkyi comprising O and/or N; and
R19, R19' and R19" are independently selected from hydrogen; Ci_6alkyl; and phenyl.
10. The compound or pharmaceutically acceptable salt according to claim 9, wherein
R1 and R9 are hydrogen;
R4 and R11 are independently selected from hydrogen, Br, CI, F; unsubstituted Ci_ 6alkyl or halo- or alkyl-substituted Ci_6alkyl; and phenyl;
R5 and R6 are independently selected from hydrogen, halogen; unsubstituted Ci_ 6alkyl and alkyl-substituted Ci_6alkyl;
R12 is selected from unsubstituted Ci-ealkyl or Ci_6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, heterocycloalkyi, heteroaryl, C3-8cycloalkyl and C2-6alkenyl. and
R19, R19' and R19" are independently selected from hydrogen; Ci-6alkyl.
1 1 . The compound or pharmaceutically acceptable salt according to any preceding claim, wherein
R4 and R11 are independently selected from hydrogen, Br, CI, and F.
12. Thecompound or pharmaceutically acceptable salt according to any of the preceding claims, wherein:
R10 is COOH;
R12 is independently selected from hydrogen; unsubstituted Ci_6alkyl or Ci-6alkyl substituted with one or more substituents independently selected from halo, hydroxy, alkoxy, heterocycloalkyi, heteroaryl, wherein the heteroaryl is a monocyclic heteroaryl comprising O and/or N; and Ci-6alkylheterocycloalkyl, wherein the heterocycloalkyi is a monocyclic heterocycloalkyi comprising O and/or N; and
R19 is hydrogen or Ci-6alkyl.
13. The compound or pharmaceutically acceptable salt according to any of the preceding claims, wherein:
R1, R4, R5, R8, R9 and R11 is each hydrogen;
R2 and R3 are independently selected from hydrogen; hydroxy; alkoxy; halogen; cyano; NR20 R21 ; COR19; COOR19; CONR20 R21 ; pyrrolidinyl; unsubstituted Ci-6alkyl or halo- or alkyl-substituted Ci_6alkyl; Ci-6alkylCOOR19; -B(OH)2; and -OR12; 0
R7 and R8 are independently selected from halogen; cyano; unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci_6alkyl; or R7 and R8 together form a 3- to 8-membered cycloalkyl ring optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, OR23, R17, R17', R18 and/or R18'; or R7 and R8 together form a 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxo, halogen, hydroxy, alkoxy, amino, cyano, R17 , R18', R17 and/or R18', wherein a heteroatom in the heterocycloalkyi ring is selected from O and N;'
R10 is COOH; and
R12 is independently selected from unsubstituted Ci_6alkyl or halo- or alkyl-substituted Ci-6alkyl.
14. The compound according to any preceding claim:
R2 and R3 are independently OR12;
R5 and R6 together form a 3- to 8-membered ring, optionally substituted with R13 , R14, R13' and/or R14' ;
R10 is COOH;
R11 is hydrogen;
R12 is unsubstituted Ci_6alkyl;
R13 and R14 or R13' and R14' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-8cycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein the heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; and
R19 is hydrogen or a pharmaceutically acceptable salt thereof.
15. The compound or pharmaceutically acceptable salt according to any preceding claim wherein:
R2 and R3 are independently OR12;
R7 and R8 together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with R17 and R18, wherein the heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22';
R10 is COOH;
R4 and R11 are indepdently selected from hydrogen and halo;
R12 is unsubstituted Ci_6alkyl;
R17 and R18 or R17' and R18' together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyi ring, optionally substituted with oxygen, halogen, hydroxy, amino, cyano, Ci_6alkyl, C3-ecycloalkyl, C2-6alkenyl or Ci-6alkoxy, wherein the heteroatom in the heterocycloalkyi ring is selected from O, N, NR22, S, SR22 or SR22R22'; and 0
R19 is hydrogen ].
16. The compound of Formula I or pharmaceutical salt thereof according to claim 1 , wherein
R2 and R3 are independently OR12; and
R5 and R6 together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyl ring, optionally substituted with R13 , R14, R13' and/or R14', wherein the heteroatom in the heterocycloalkyl ring is selected from O, N, NR22, S, SR22 or SR22R22'
17. The compound or pharmaceutical salt thereof according to claim 3,
wherein
R2 and R3 are independently OR12; and
R7 and R8 together form a 3- to 8-membered cycloalkyl ring or 3- to 8- membered heterocycloalkyl ring, optionally substituted with R17, R18, R17' and/or R18', wherein the heteroatom in the heterocycloalkyl ring is selected from O, N, NR22, S, SR22 or SR22R22'.
18. A compound from Table 1 A or a pharmaceutically acceptable salt thereof.
19 A compound from Table 1 B or a pharmaceutically acceptable salt thereof.
20. A method of treating or preventing a virus infection in a subject susceptible to or suffering from the virus infection comprising administering to the subject an inhibitor of a HBe or HBs antigen wherein the inhibitor is a compound of Formula I, Formula IA or Formula IB according to any one of claims 1 -19.
21 . A method of treating or preventing a hepatitis B virus infection in a subject susceptible to or suffering from the hepatitis B virus infection, comprising administering to the subject an inhibitor of a HBe or HBs antigen wherein the inhibitor is a compound of Formula I, Formula IA or Formula IB according to any one of claims 1 -19.
22. A method of treating or preventing a hepatitis B virus infection in a subject susceptible to or suffering from the hepatitis B virus infection comprising administering to the subject an inhibitor of a HBe or HBs antigen wherein the inhibitor comprises a compound from Table 1 A or Table 1 B
23. A method according to claim 22, wherein the inhibitor comprises a compound from 0
Table 1A.
24. A method of inhibiting levels of HBe or HBs antigens in a mammal, comprising administering to said mammal a therapeutically effective amount of a compound of Formula I, Formula IA or Formula IB selected from Table 1 A and Table 1 B, or a pharmaceutically acceptable salt, solvate or hydrate thereof.
25. A method according to claim 24, wherein the mammal is a human.
26. A pharmaceutical composition comprising a pharmaceutically acceptable diluent and a therapeutically effective amount of a compound as defined in Formula I, Formula IA or Formula IB according to any of claims 1 -19.
27. A compound of Formula I, Formula IA or Formula IB according to any of claims 1 -19, or a pharmaceutical composition according to claim 26, for use in therapy.
28. A compound of Formula I, Formula IA or Formula IB according to any of claims 1 -19, or a pharmaceutical composition according to claim 26, for use in the treatment of a viral infection.
29. A compound according to claim 28 for use in the treatment of a viral infection, wherein the viral infection is a hepatitis B viral infectio.
30. Use of a compound of Formula I, Formula IA or Formula IB according to any of claims 1 -19, 28 or 29 or a pharmaceutical composition according to claim 26, in the manufacture of a medicament for use in the treatment of a hepatitis B viral infection in a human.
PCT/IB2018/051080 2017-02-21 2018-02-21 Dihydroquinolizinones as antivirals Ceased WO2018154466A1 (en)

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