HK1209427B

HK1209427B - Azaheterocycles as bir2 and/or bir3 inhibitors

Info

Publication number: HK1209427B
Application number: HK15110299.0A
Authority: HK
Inventors: Joan Heather Hogg; Kang Le; Yan Lou; Steven Gregory Mischke; Stacy Remiszewski
Original assignee: 霍夫曼-拉罗奇有限公司
Priority date: 2012-08-15
Filing date: 2013-08-06
Publication date: 2018-05-04

Description

Aza-heterocycles as BIR2 and/or BIR3 inhibitors

Technical Field

The present invention relates to substituted isoindolines and tetrahydro-isoquinolines as inhibitors of SMAC protein binding to Inhibitors of Apoptotic Proteins (IAPs), and/or activated caspase (caspase) protein binding to IAPs. These molecules are useful for ameliorating, treating or controlling cancer, particularly solid tumors.

These compounds bind to BIR2 and/or BIR3 regions of IAP proteins (including XIAP and cIAP) resulting in the activation or reactivation of the caspase cascade, and are therefore useful in the treatment of proliferative diseases, including cancer.

Background

Cancer is a disease of uncontrolled cell growth, which leads to local enlargement of the tumor and possibly distant metastasis. One mechanism of cancer cell growth is by avoiding apoptosis or programmed cell death. Alterations in apoptotic pathways have been linked to cancer cell tolerance to standard therapies (e.g., chemotherapy or radiation), and to cancer development and progression. See, e.g., E.dean et al, "X-linked inhibitor of apoptosis protein as a therapeutic target," Expert Optin. Ther. targets (2007)11(11): 1459-.

The two basic pathways of apoptotic cell death are the internal and external pathways. The internal apoptotic pathway can be initiated by a variety of mechanisms including cellular stress and drug-induced DNA damage. The external pathway may be initiated by chemokine activation of death receptors. Initiation of either pathway results in activation of a family of proteases known as caspases. Once activated, the caspase can cleave multiple substrates, creating a cascade of reaction events, further leading to activation of effector caspases 3 and 7, and ultimately cell death. The IAP family of proteins can bind and inhibit the activity of caspases, thus inhibiting apoptosis. See, e.g., Dean, supra, 1460.

IAPs can include up to three identical domains, called Baculovirus IAP Repeat (BIR) domains: BIR1, BIR2 and BIR 3. The BIR3 domain of typical IAPs (cIAP and XIAP) can bind to and inhibit activated caspase 9. In contrast, the BIR2 domain binds to and inhibits caspases 3 and 7. Pro-apoptotic protein Smac (also known as DIABLO) can block BIR2 and BIR3 domains of IAPs from competing with activated caspases, resulting in the release of the activated caspases from the IAPs and completion of the apoptotic program. See, e.g., S.Wang, "Design of Small-Mobile Smac metrics as IAP oligonucleotides," Current diagnostics in Microbiology and diagnostics 348, DOI 10.1007/82_2010_111, pp.89-113.

Peptides and small molecules have been reported to bind to the BIR3 region of XIAP and cIAP, mimicking the effect of Smac proteins, releasing activated caspases that are inhibited by IAPs. See, e.g., Dean, supra; and M.Gyrd-Hanse et al, "IAPs From caspase inhibitors to modulators of NF-. kappa.B, inflammation and Cancer," Nature Review/Cancer, August 2010, Vol10: 561-.

Summary of The Invention

One aspect of the present invention is a compound of formula I or a pharmaceutically acceptable salt thereof

Wherein R is¹、R²、R³M, n and q are as described in the present application.

The invention also relates to pharmaceutical compositions comprising one or more compounds of the invention, or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier or excipient.

The present invention also relates to a method of alleviating, controlling or treating cancer, including particularly solid tumors, such as lung, pancreatic, colon, breast, bone and prostate cancer, in a mammal, particularly a human, comprising administering to said mammal a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof.

Detailed Description

Definition of

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

The following terms used in the present application have the following definitions.

"alkyl" refers to a monovalent straight or branched chain saturated hydrocarbon group having 1 to 12 carbon atoms. In particular embodiments, the alkyl group has 1 to 6 carbon atoms, and in more particular embodiments, the alkyl group has 1 to 4 carbon atoms. As used herein, "lower alkyl" refers to an alkyl (C) group having 1 to 6 carbon atoms_1-6-an alkyl group). Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl (also known as n-butyl), isobutyl, sec-butyl, tert-butyl, pentyl, hexyl and the like. The alkyl group may optionally be enriched with deuterium, e.g. -CD₃、-CD₂CD₃And the like.

"aryl" refers to a monovalent aromatic carbocyclic mono-, bi-, or tricyclic ring system containing from 6 to 19 carbon ring atoms. Examples of aryl moieties include, but are not limited to, phenyl, naphthyl, tolyl, xylyl, pyridyl, quinolinyl, pyrimidinyl, imidazolyl, thiazolyl, anthracenyl, tetrazolyl, and fluorenyl. A specific aryl group is phenyl.

"cycloalkyl" refers to a substituted or unsubstituted stable monovalent saturated monocyclic, bicyclic, or tricyclic ring system consisting of 3 to 10 ring carbon atoms. In a specific embodiment, cycloalkyl refers to a ring having 3 to 8 ring carbon atoms, more specifically 3 to 7 carbon atoms ("C)_3-7-cycloalkyl ") monovalent saturated monocyclic hydrocarbon radicals. Specific cycloalkyl groups are monocyclic cycloalkyl groups. Examples of monocyclic cycloalkyl are cyclopropyl, cyclobutyl, cyclopentylPhenyl, cyclohexyl or cycloheptyl. Bicyclic means consisting of two saturated carbocyclic rings having one or more common carbon atoms. An example of bicycloalkyl is bicyclo [2.2.1]Heptyl or bicyclo [2.2.2]And (4) octyl. Tricyclic means consisting of three saturated carbocyclic rings having one or more common carbon atoms. Examples of tricyclic cycloalkyl groups include adamantane.

When two or more rings are involved, such as aryl fused with cycloalkyl, "fused" refers to rings having at least two atoms in common. An example of aryl fused to cycloalkyl is tetrahydronaphthyl.

"halogen" or "halo" refers to an atom selected from F, Cl, Br or I. In particular embodiments, halogen refers to F and Cl.

"heteroatom" means an atom selected from N, O or S.

"heteroaryl" refers to a substituted or unsubstituted aromatic heterocyclic ring system containing up to two rings, wherein at least one ring contains 1,2, or 3 heteroatoms and the remaining ring atoms are carbon. Examples of heteroaryl groups include, but are not limited to, thienyl, furyl, indolyl, pyrrolyl, pyridyl, pyrazinyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, quinolinyl, isoquinolinyl, indazolyl, pyrimidinyl, imidazolyl, triazolyl, tetrazolyl, triazinyl, pyrazolyl, benzo [ d ] isoxazolyl, 2-oxo-2H-chromen (chromen) -4-yl, benzo [ d ] isoxazolyl, benzo [ b ] thienyl, naphthyridinyl, and cinnolinyl.

Where the heteroaryl group is bicyclic, it is understood that one ring can be aryl and the other ring heteroaryl, and both rings can be independently substituted or unsubstituted.

"Heterocyclyl" or "heterocycle" refers to a substituted or unsubstituted monovalent saturated or partially unsaturated mono-or bicyclic non-aromatic hydrocarbon system having from 3 to 9 ring atoms, including 1,2, or 3 ring heteroatoms selected from N, O and S, with the remaining ring atoms being carbon. In particular embodiments, heterocycloalkyl is a monovalent saturated monocyclic ring system having 4 to 7 ring atoms, including 1,2, or 3 ring heteroatoms selected from N, O and S, with the remaining ring atoms being carbon. Examples of monocyclic saturated heterocycloalkyl groups are aziridinyl, oxetanyl, azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydrofuranyl, piperazinyl, morpholinyl, thiomorpholinyl, 1-dioxo-thiomorpholin-4-yl, azepanyl, diazepanyl, homopiperazinyl or oxazepanyl. Examples of partially unsaturated heterocycloalkyl groups are dihydrofuranyl, imidazolinyl, dihydro-oxazolyl, dihydro-oxadiazolyl, dihydro-triazolyl, tetrahydro-pyridyl, tetrahydro-triazinyl or dihydropyranyl.

Where the heterocyclyl is bicyclic, it is understood that one ring may be heterocyclyl and the other ring cycloalkyl, and either or both rings may be independently substituted. Examples of bicyclic saturated heterocycloalkyl are 8-aza-bicyclo [3.2.1] octyl, quinuclidinyl, 8-oxa-3-aza-bicyclo [3.2.1] octyl, 9-aza-bicyclo [3.3.1] nonyl, 3-oxa-9-aza-bicyclo [3.3.1] nonyl or 3-thia-9-aza-bicyclo [3.3.1] nonyl.

“IC₅₀By "is meant the concentration of a particular compound required to inhibit 50% of a particular measured activity. IC (integrated circuit)₅₀Can be measured specifically as in example 48 that follows.

"oxo" or ("oxy") refers to ═ O.

By "pharmaceutically acceptable" (e.g., pharmaceutically acceptable carrier, excipient, etc.) is meant pharmacologically acceptable and substantially non-toxic to the subject to which the particular compound is administered.

"pharmaceutically acceptable salt" refers to conventional acid addition salts or base addition salts that retain the biological effects and properties of the compounds of the present invention and are formed from suitable non-toxic organic or inorganic acids or organic or inorganic bases. Examples of acid addition salts include those derived from inorganic acids such as hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfamic, phosphoric and nitric acids, and those derived from organic acids such as p-toluenesulfonic, salicylic, methanesulfonic, oxalic, succinic, citric, malic, lactic, fumaric, trifluoroacetic acids, and the like. Examples of base addition salts include those derived from ammonium, potassium, sodium, and quaternary ammonium hydroxides (e.g., tetramethylammonium hydroxide). Chemical modifications to convert a pharmaceutical compound (i.e., drug) into a salt are well known to pharmaceutical chemists to achieve improved physical and chemical stability, hygroscopicity, flowability, and solubility of the compound. See, e.g., Ansel et al, Pharmaceutical document Forms and Drug Delivery Systems (1995) atpgs.456-457.

"substituted" in, for example, substituted alkyl, aryl, or heteroaryl means that substitution (i.e., replacement of one hydrogen atom) may occur at one or more positions, and that the substituents at each substitution position are independently selected from the indicated options, unless otherwise indicated. The term "optionally substituted" refers to the following: one or more hydrogen atoms of a chemical group (having one or more hydrogen atoms) may, but need not, be substituted with another substituent.

The definitions set forth herein apply regardless of whether the terms appear alone or in combination. It is contemplated that the definitions described herein may be concatenated to form chemically related combinations such as "heterocycloalkylaryl", "haloalkylheteroaryl", or "arylalkyl heterocycloalkyl". The last member of the combination is the group attached to the rest of the molecule. The other members of the combination are attached to the linker in reverse order of the literal order, e.g., an arylalkyl-heterocycloalkyl combination means that the heterocycloalkyl group is substituted with alkyl and the alkyl group is substituted with aryl.

As used herein, a missing substituent is considered to be H if a formula or group appears to have one missing substituent, i.e., the valence is shown to be incomplete.

In the structural formulae presented herein, broken bond (a) indicates that the substituent is located below the paper surface, while wedge bond (b) indicates that the substituent is located above the paper surface.

In one embodiment, the invention relates to compounds of formula I

Wherein:

R¹selected from H and halogen;

R²is selected from

Aryl, which may optionally be substituted by lower alkyl, OR⁴And a substitution of a halogen,

an aryl radical, which is fused to a cycloalkyl radical, and

heteroaryl, which may be optionally substituted by lower alkyl;

R³is selected from

Lower alkyl, which may optionally be OR⁴And an aryl group for substitution, and a pharmaceutically acceptable salt thereof,

a cycloalkyl group,

a heterocyclic group, and

an aryl group;

R⁴selected from H and lower alkyl;

n is 1 or 2;

m is 0 or 1; and is

q is 0,1 or 2;

or a pharmaceutically acceptable salt thereof.

In one embodiment, the invention relates to compounds of formula I:

wherein:

R¹selected from H and halogen;

R²is selected from

Aryl, which may optionally be substituted by C_1-6-alkyl, OR⁴And a substitution of a halogen,

aryl, with C_3-7-cycloalkyl fused, and

heteroaryl, which may optionally be substituted by C_1-6-alkyl substitution;

R³is selected from

·C_1-6-alkyl, which may optionally be OR⁴And an aryl group for substitution, and a pharmaceutically acceptable salt thereof,

·C_3-7-a cycloalkyl group,

a heterocyclic group, and

an aryl group;

R⁴selected from H and C_1-6-an alkyl group;

n is 1 or 2;

m is 0 or 1; and is

q is 0,1 or 2;

or a pharmaceutically acceptable salt of the above compound.

In one embodiment, the present invention relates to a compound described herein, or a pharmaceutically acceptable salt thereof, wherein R¹Is halogen.

In one embodiment, the invention relates to compounds described herein, wherein R is¹Is H.

In one embodiment, the present invention relates to a compound described herein, or a pharmaceutically acceptable salt thereof, wherein R²Is aryl, which may optionally be OR⁴Halogen and C_1-6-alkyl substitution.

In one embodiment, the present invention relates to a compound described herein, or a pharmaceutically acceptable salt thereof, wherein R²Is phenyl or naphthyl.

In one embodiment, the present invention relates to a compound described herein, or a pharmaceutically acceptable salt thereof, wherein R²Is heteroaryl, which may optionally be substituted by C_1-6-alkyl substitution.

In one embodiment, the invention relates to compounds described herein, wherein R is²Selected from quinolyl, benzo [ b ]]Thienyl or indolyl.

In one embodiment, the present invention relates to a compound described herein, or a pharmaceutically acceptable salt thereof, wherein R³Is C_1-6-alkyl, which may optionally be OR⁴And aryl substitution.

In one embodiment, the present invention relates to a compound described herein, or a pharmaceutically acceptable salt thereof, wherein R³Is C substituted by phenyl_1-6-an alkyl group.

In one embodiment, the present invention relates to a compound described herein, or a pharmaceutically acceptable salt thereof, wherein R³Is OR⁴And R is⁴Is H.

In one embodiment, the present invention relates to a compound described herein, or a pharmaceutically acceptable salt thereof, wherein R³Is C_3-7-a cycloalkyl group.

In one embodiment, the present invention relates to a compound described herein, or a pharmaceutically acceptable salt thereof, wherein R³Cyclohexyl or cyclopentyl.

In one embodiment, the present invention relates to a compound described herein, or a pharmaceutically acceptable salt thereof, wherein R³Is an aryl group.

In one embodiment, the invention relates to compounds described herein, wherein R is³Is phenyl.

In one embodiment, the present invention relates to a compound described herein, or a pharmaceutically acceptable salt thereof, wherein R³Is a heterocyclic group.

In one embodiment, the invention relates to compounds described herein, wherein R is³Is tetrahydropyranyl.

In one embodiment, the present invention relates to a compound described herein, or a pharmaceutically acceptable salt thereof, wherein m is 0, n is 1, and q is 0.

In one embodiment, the invention relates to compounds described herein, wherein R is¹Is H, R²Is aryl, R³Is C_1-6-alkyl, n is 1 and m and q are 0.

In one embodiment, the invention relates to compounds described herein, wherein R is³Is C_1-6-an alkyl group, n is 1 and m is 0, said compound being selected from:

(S) -2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride;

((S) -N- (2, 6-dichlorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) isoindoline-1-carboxamide hydrochloride;

(S) -N- (2-chloro-6-fluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide hydrochloride;

(S) -N- (benzo [ b ] thiophen-4-yl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxylic acid amide hydrochloride;

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) pentanoyl) isoindoline-1-carboxamide hydrochloride;

(S) -N- (2, 6-difluorophenyl) -2- ((S) -3, 3-dimethyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxylic acid amide hydrochloride;

2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) -N-phenylisoindoline-1-carboxamide hydrochloride;

n- (2-methoxyphenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide hydrochloride;

n-benzyl-2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide hydrochloride;

2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) -N-phenethylisoindoline-1-carboxamide hydrochloride;

(S) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) -N- (naphthalen-1-yl) isoindoline-1-carboxylic acid amide hydrochloride;

(R) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) -N- (naphthalen-1-yl) isoindoline-1-carboxylic acid amide hydrochloride;

(S) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) -N- (naphthalen-2-yl) isoindoline-1-carboxylic acid amide hydrochloride;

(S) -N- (2-fluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide hydrochloride;

(S) -N- (2-chlorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide hydrochloride;

(S) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) -N- (5,6,7, 8-tetrahydronaphthalen-1-yl) isoindoline-1-carboxamide hydrochloride;

(S) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) -N- (quinolin-8-yl) isoindoline-1-carboxamide hydrochloride;

n- (isoquinolin-1-yl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide hydrochloride;

(S) -N- (1-methyl-1H-indol-4-yl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide hydrochloride;

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -3-methoxy-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide hydrochloride;

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3R) -3-hydroxy-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide hydrochloride;

(S) -N- (2, 6-difluorophenyl) -2- ((S) -4-methyl-2- ((S) -2- (methylamino) propionylamino) pentanoyl) isoindoline-1-carboxamide hydrochloride;

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -2- ((S) -2- (methylamino) propionylamino) -3-phenylbutyryl) isoindoline-1-carboxamide hydrochloride; and

(S) -N- (2-fluoro-6-methylphenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide hydrochloride;

or a pharmaceutically acceptable salt of any of the foregoing.

In one embodiment, the invention relates to compounds described herein, wherein R is³Is C_1-6-alkyl, n is 2 and m is 0, said compound being selected from:

(S) -2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2-chloro-6-fluoro-phenyl) -amide hydrochloride;

6-chloro-2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide;

6-fluoro-2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide;

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) pentanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride;

(S) -N- (2, 6-difluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride;

(S) -N- (2, 6-dichlorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride;

(S) -N- (2, 6-difluorophenyl) -2- ((S) -3, 3-dimethyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride;

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3R) -3-methoxy-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride;

n- (2, 6-difluorophenyl) -7-fluoro-2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide;

n- (2, 6-difluorophenyl) -6-fluoro-2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide;

7-chloro-N- (2, 6-difluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide;

7-chloro-N- (2, 6-difluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide; and

6-chloro-N- (2, 6-difluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide;

or a pharmaceutically acceptable salt of any of the foregoing.

In one embodiment, the invention relates to compounds described herein, wherein R is³Is C_1-6-alkyl, n is 1 and m is 1, said compound being

(S) -N- (2, 6-difluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-3-carboxamide;

or a pharmaceutically acceptable salt thereof.

In one embodiment, the invention relates to compounds described herein, wherein R is³Is C_3-7-cycloalkyl and n is 1, said compound being selected from:

(S) -2- [ (S) -2-cyclohexyl-2- ((S) -2-methylamino-propionylamino) -acetyl ] -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride; and

(S) -2- ((S) -2-cyclopentyl-2- ((S) -2- (methylamino) propionylamino) acetyl) -N- (2, 6-difluorophenyl) isoindoline-1-carboxylic acid amide hydrochloride;

or a pharmaceutically acceptable salt of any of the foregoing.

In one embodiment, the invention relates to compounds described herein, wherein R is³Is C_3-7-cycloalkyl and n is 2, said compound being selected from:

(S) -2- [ (S) -2-cyclohexyl-2- ((S) -2-methylamino-propionylamino) -acetyl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride; and

(S) -2- ((S) -2-cyclopentyl-2- ((S) -2- (methylamino) propionylamino) acetyl) -N- (2, 6-difluorophenyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride;

or a pharmaceutically acceptable salt of any of the foregoing.

In one embodiment, the invention relates to compounds described herein, wherein R is³Is a heterocyclic group, said compound being selected from:

(S) -N- (2, 6-difluorophenyl) -2- ((S) -2- (methylamino) propionylamino) -2- (tetrahydro-2H-pyran-4-yl) acetyl) isoindoline-1-carboxylic acid amide hydrochloride; and

(S) -N- (2, 6-difluorophenyl) -2- ((S) -2- (methylamino) propionylamino) -2- (tetrahydro-2H-pyran-4-yl) acetyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride;

or a pharmaceutically acceptable salt of any of the foregoing.

In one embodiment, the invention relates to compounds described herein, wherein R is³Is aryl, the compound is

(S) -N- (2, 6-difluorophenyl) -2- ((S) -2- (methylamino) propionylamino) -2-phenylacetyl) isoindoline-1-carboxamide hydrochloride;

or a pharmaceutically acceptable salt of the above compound.

In one embodiment, the present invention relates to a compound described herein selected from the group consisting of:

((S) -2- [ (S) -2-cyclohexyl-2- ((S) -2-methylamino-propionylamino) -acetyl ] -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride;

(S) -N- (2, 6-difluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide hydrochloride;

(S) -N- (2, 6-dichlorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) isoindoline-1-carboxamide hydrochloride;

(S) -N- (2, 6-difluorophenyl) -2- ((S) -2- (methylamino) propionylamino) -2- (tetrahydro-2H-pyran-4-yl) acetyl) isoindoline-1-carboxylic acid amide hydrochloride;

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) pentanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride; and

or a pharmaceutically acceptable salt of each of the foregoing compounds.

In one embodiment, the present invention relates to a pharmaceutical composition comprising as an active ingredient any of the compounds described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

In one embodiment, the invention relates to compounds as described herein for use as therapeutically active substances.

In one embodiment, the present invention relates to the use of a compound described herein, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the therapeutic and/or prophylactic treatment of cancer.

In one embodiment, the present invention relates to the use of a compound described herein, or a pharmaceutically acceptable salt thereof, in the therapeutic and/or prophylactic treatment of cancer.

In one embodiment, the invention relates to a method of treating or ameliorating cancer comprising administering to a subject in need of such treatment a therapeutically effective amount of a compound described herein.

In one embodiment, the invention relates to compounds of formula I

Wherein:

R¹selected from H, Cl and F;

R²is selected from

A phenyl group, in which the phenyl group is,

phenyl, which is selected from F, Cl, -CH₃and-OCH₃Is substituted once or twice with the substituent(s),

a naphthyl group,

5,6,7, 8-tetrahydronaphthyl,

a benzo [ b ] thienyl group,

a quinolyl group, which is a substituent of the quinoline group,

isoquinolinyl, and

indolyl, which may optionally be substituted by-CH₃Substitution;

R³is selected from

The isopropyl group is a group represented by,

a tert-butyl group,

·CH₃-CH₂-C(H,CH₃)-，

·CH(CH₃)₂-CH₂-，

·CH₃-CH₂-C(H,OCH₃)-，

·CH₃-C(H,OH)-，

·CH₃-C(H,OCH₃)-，

a phenyl group, in which the phenyl group is,

phenyl-C (H, CH)₃)-，

tetrahydro-2H-pyranyl radical,

cyclopentyl, and

cyclohexyl;

R⁴selected from H and lower alkyl;

n is 1 or 2;

m is 0 or 1; and is

q is 0,1 or 2;

or a pharmaceutically acceptable salt thereof.

In one embodiment, the invention relates to compounds of formula I

Wherein:

R¹is H;

R²is selected from

A phenyl group, in which the phenyl group is,

and

a naphthyl group,

R³is selected from

The isopropyl group is a group represented by,

a tert-butyl group,

·CH₃-CH₂-C(H,CH₃)-，

·CH(CH₃)₂-CH₂-，

·CH₃-CH₂-C(H,OCH₃)-，

·CH₃-C (H, OH) -, and

·CH₃-C(H,OCH₃)-；

R⁴selected from H and lower alkyl;

n is 1;

m is 0; and is

q is 0;

or a pharmaceutically acceptable salt thereof.

One embodiment of the present invention is directed to compounds of formula I, or pharmaceutically acceptable salts thereof, wherein R¹Is halogen. In specific embodiments, R¹Is F or Cl.

Another embodiment of the present invention is directed to compounds of formula I, or pharmaceutically acceptable salts thereof, wherein R¹Is H.

Another embodiment of the present invention is directed to compounds of formula I, or pharmaceutically acceptable salts thereof, wherein R²Is aryl, which may optionally be OR⁴Halogen and lower alkyl. In specific embodiments, R²Is phenyl or naphthyl, each of which may be optionally substituted as defined above.

Another embodiment of the present invention is directed to compounds of formula I, or pharmaceutically acceptable salts thereof, wherein R²Is heteroaryl, which may be optionally substituted with lower alkyl. In specific embodiments, R²Selected from quinolyl, benzo [ b ]]Thienyl or indolyl.

Another embodiment of the present invention is directed to compounds of formula I, or pharmaceutically acceptable salts thereof, wherein R³Lower alkyl, which may be optionally OR⁴And aryl substitution. In specific embodiments, R³Is lower alkyl substituted by phenyl. In other embodiments, R⁴Is H.

Another embodiment of the present invention is directed to compounds of formula I, or pharmaceutically acceptable salts thereof, wherein R³Is a cycloalkyl group. In specific embodiments, R³Cyclohexyl or cyclopentyl.

Another embodiment of the present invention is directed to compounds of formula I, or pharmaceutically acceptable salts thereof, wherein R³Is an aryl group. In specific embodiments, R³Is phenyl.

Another embodiment of the present invention is directed to compounds of formula I, or pharmaceutically acceptable salts thereof, wherein R³Is a heterocyclic group. In specific embodiments, R³Is tetrahydropyranyl.

Another embodiment of the present invention relates to compounds of formula I, or pharmaceutically acceptable salts thereof, wherein m is 0.

Another embodiment of the present invention is directed to compounds of formula I, or pharmaceutically acceptable salts thereof, wherein n is 1.

Another embodiment of the present invention is directed to compounds of formula I, or pharmaceutically acceptable salts thereof, wherein n is 2.

Another embodiment of the present invention is directed to compounds of formula I, or pharmaceutically acceptable salts thereof, wherein q is 0.

Another embodiment of the invention relates to compounds of formula I, wherein R¹Is H, R²Is aryl, R³Is lower alkyl, n is 1 and m and q are 0.

Compounds of the invention (wherein R³Is alkyl, which may be optionally substituted as defined above, n is 1, and m is 0) includes:

(S) -2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride (example 1);

((S) -N- (2, 6-dichlorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) isoindoline-1-carboxamide hydrochloride (example 7);

(S) -N- (2-chloro-6-fluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) isoindoline-1-carboxamide hydrochloride (example 8);

(S) -N- (benzo [ b ] thiophen-4-yl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) isoindoline-1-carboxamide hydrochloride (example 9);

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) pentanoyl) isoindoline-1-carboxamide hydrochloride (example 10);

(S) -N- (2, 6-difluorophenyl) -2- ((S) -3, 3-dimethyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) isoindoline-1-carboxylic acid amide hydrochloride (example 11);

2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) -N-phenylisoindoline-1-carboxamide hydrochloride (example 16);

n- (2-methoxyphenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide hydrochloride (example 17);

n- (2-methoxyphenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide hydrochloride (example 18);

n-benzyl-2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide hydrochloride (example 19);

2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) -N-phenethylisoindoline-1-carboxamide hydrochloride (example 20);

(S) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -N- (naphthalen-1-yl) isoindoline-1-carboxamide hydrochloride (example 21);

(R) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -N- (naphthalen-1-yl) isoindoline-1-carboxamide hydrochloride (example 22);

(S) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -N- (naphthalen-2-yl) isoindoline-1-carboxamide hydrochloride (example 23);

(S) -N- (2-fluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) isoindoline-1-carboxamide hydrochloride (example 24);

(S) -N- (2-chlorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) isoindoline-1-carboxamide hydrochloride (example 26);

(S) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -N- (5,6,7, 8-tetrahydronaphthalen-1-yl) isoindoline-1-carboxamide hydrochloride (example 27);

(S) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -N- (quinolin-8-yl) isoindoline-1-carboxamide hydrochloride (example 28);

n- (isoquinolin-1-yl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) isoindoline-1-carboxamide hydrochloride (example 29);

(S) -N- (1-methyl-1H-indol-4-yl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) isoindoline-1-carboxamide hydrochloride (example 30);

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -3-methoxy-2- ((S) -2- (methylamino) propionylamino) butanoyl) isoindoline-1-carboxamide hydrochloride (example 32);

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3R) -3-hydroxy-2- ((S) -2- (methylamino) propionylamino) butanoyl) isoindoline-1-carboxamide hydrochloride (example 33);

(S) -N- (2, 6-difluorophenyl) -2- ((S) -4-methyl-2- ((S) -2- (methylamino) propionylamino) pentanoyl) isoindoline-1-carboxamide hydrochloride (example 35);

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -2- ((S) -2- (methylamino) propionylamino) -3-phenylbutyryl) isoindoline-1-carboxamide hydrochloride (example 40); and

(S) -N- (2-fluoro-6-methylphenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) isoindoline-1-carboxamide hydrochloride (example 47);

or a pharmaceutically acceptable salt of any of the foregoing.

Compounds of the invention (wherein R³Is alkyl, which may be optionally substituted as defined above, n is 2 and m is 0) includes:

(S) -2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2-chloro-6-fluoro-phenyl) -amide hydrochloride (example 4);

6-chloro-2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide (example 5);

6-fluoro-2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide (example 6);

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) pentanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride (example 15);

(S) -N- (2, 6-difluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride (example 25);

(S) -N- (2, 6-dichlorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride (example 36);

(S) -N- (2, 6-difluorophenyl) -2- ((S) -3, 3-dimethyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride (example 38);

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3R) -3-methoxy-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride (example 39);

n- (2, 6-difluorophenyl) -7-fluoro-2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide (example 41);

n- (2, 6-difluorophenyl) -7-fluoro-2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide (example 42);

n- (2, 6-difluorophenyl) -6-fluoro-2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide (example 43);

7-chloro-N- (2, 6-difluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide (example 44);

7-chloro-N- (2, 6-difluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide (example 45); and

6-chloro-N- (2, 6-difluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide (example 46);

or a pharmaceutically acceptable salt of any of the foregoing.

Compounds of the invention (wherein R³Is alkyl, which may be optionally substituted as defined above, n is 1 and m is 1) includes:

(S) -2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -1,2,3, 4-tetrahydro-isoquinoline-3-carboxylic acid (2, 6-difluoro-phenyl) -amide (example 31);

or a pharmaceutically acceptable salt thereof.

Compounds of the invention (wherein R³Is cycloalkyl and n is 1) comprises:

(S) -2- [ (S) -2-cyclohexyl-2- ((S) -2-methylamino-propionylamino) -acetyl ] -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride (example 2); and

(S) -2- ((S) -2-cyclopentyl-2- ((S) -2- (methylamino) propionylamino) acetyl) -N- (2, 6-difluorophenyl) isoindoline-1-carboxamide hydrochloride (example 13);

or a pharmaceutically acceptable salt of any of the foregoing.

Compounds of the invention (wherein R³Is cycloalkyl and n is 2) comprises:

(S) -2- [ (S) -2-cyclohexyl-2- ((S) -2-methylamino-propionylamino) -acetyl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride (example 3); and

(S) -2- ((S) -2-cyclopentyl-2- ((S) -2- (methylamino) propionylamino) acetyl) -N- (2, 6-difluorophenyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride (example 37);

or a pharmaceutically acceptable salt of any of the foregoing.

Compounds of the invention (wherein R³Is a heterocyclic group and n is 1) includes:

(S) -N- (2, 6-difluorophenyl) -2- ((S) -2- ((S) -2- (methylamino) propionylamino) -2- (tetrahydro-2H-pyran-4-yl) acetyl) isoindoline-1-carboxylic acid amide hydrochloride (example 12);

or a pharmaceutically acceptable salt thereof.

Compounds of the invention (wherein R³Is heterocyclyl and n is 2)Comprises the following steps:

(S) -N- (2, 6-difluorophenyl) -2- ((S) -2- ((S) -2- (methylamino) propionylamino) -2- (tetrahydro-2H-pyran-4-yl) acetyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride (example 14);

or a pharmaceutically acceptable salt of the above compound.

Compounds of the invention (wherein R³Is aryl and n is 1) comprises:

(S) -N- (2, 6-difluorophenyl) -2- ((S) -2- ((S) -2- (methylamino) propionylamino) -2-phenylacetyl) isoindoline-1-carboxamide hydrochloride (example 34);

or a pharmaceutically acceptable salt of the above compound.

Another embodiment of the present invention relates to the following compounds:

((S) -2- [ (S) -2-cyclohexyl-2- ((S) -2-methylamino-propionylamino) -acetyl ] -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride (example 2);

(S) -2- [ (S) -2-cyclohexyl-2- ((S) -2-methylamino-propionylamino) -acetyl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride (example 3);

(S) -N- (2, 6-dichlorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) isoindoline-1-carboxamide hydrochloride (example 7);

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) pentanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride (example 15); and

or a pharmaceutically acceptable salt of any of the foregoing.

The compounds of formula I and their salts have at least one asymmetric carbon atom and may therefore exist as mixtures of different stereoisomers. The various isomers can be separated by known separation methods, such as chromatography.

The compounds disclosed herein and encompassed by formula I above may exhibit tautomerism or structural isomerism. The present invention is intended to encompass any tautomeric or structurally isomeric form of these compounds, or mixtures of such forms, and is not limited to any one tautomeric or structurally isomeric form described in the above formula.

Dosage form

The compounds of the invention preferably bind to the BIR domain of IAPs, preventing IAPs from binding to other proteins. Examples of Bir binding proteins include, but are not limited to, caspase 3, caspase 7, caspase 9, Smac, and the like. Examples of IAPs include, but are not limited to, XIAP, cIAP1, cIAP2, or NAIP. In one aspect. The compounds of the invention bind to the BIR2 and/or BIR3 domains of XIAP, cIAP1 and/or cIAP 2. In another aspect, the compounds of the invention bind to the BIR2 domain of XIAP, cIAP1 and/or cIAP 2.

The compounds of the invention are useful for inducing apoptosis in cells or sensitizing cells, particularly cancer cells, to apoptotic signals. Apoptotic signals may be induced in cancer cells by, for example, radiation therapy or anti-tumor chemotherapy. Alternatively, apoptotic signals may be induced in cancer cells by activation of death receptors by death receptor agonists. Death receptor agonists may be of natural origin, such as tumor necrosis factor alpha (TNF- α), or of non-natural origin, such as synthetic antibodies, such as DR4 or DR5 antibodies.

The compounds of the invention are useful for alleviating, controlling or treating cell proliferative disorders, such as in particular neoplastic disorders. These compounds, and formulations containing the compounds, may be used in the treatment or control of hematological cancers, such as acute myeloid leukemia, or solid tumors, such as breast, colon, lung, and prostate cancers.

A "therapeutically effective amount" or "effective amount" of a compound of the invention refers to an amount of a compound effective to prevent, alleviate or alleviate symptoms of a disease in a subject to be treated or to prolong the survival of the subject. Determination of a therapeutically effective amount is within the skill of the art.

The therapeutically effective amount or dosage of the invention may vary within wide limits and may be determined in a manner known in the art. The dosage is adjusted in each particular case to the individual need, including the particular compound administered, the route of administration, the condition to be treated and the patient to be treated. Generally, in the case of oral or parenteral administration to an adult human weighing about 70Kg, a daily dose of about 10mg to about 10,000mg, preferably about 200mg to about 1,000mg should be appropriate, although the upper limit may be exceeded. The daily dose may be administered as a single dose or divided doses, or for parenteral administration it may be administered as one or more bolus injections or as a continuous infusion.

Pharmaceutical formulations for use in the practice of the present invention (i.e., comprising a compound of the present invention) may be administered in vivo, such as orally (e.g., in the form of tablets, coated tablets, lozenges, hard and soft gelatin capsules, solutions, emulsions or suspensions), nasally (e.g., in the form of a nasal spray), or rectally (e.g., in the form of suppositories). However, the administration can also be administered parenterally, such as intramuscularly or intravenously (e.g. in the form of injectable solutions). Furthermore, the administration may be given topically (e.g., in the form of an ointment, cream, or oil).

Composition/formulation

In an alternative embodiment, the present invention encompasses a pharmaceutical composition comprising at least one compound of formula I, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient and/or carrier.

These pharmaceutical compositions may be adapted for oral, nasal, topical (including buccal and sublingual), rectal, vaginal and/or parenteral administration. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular form of administration. The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form will generally be that amount of the compound of formula I which produces a therapeutic effect. Typically, the amount will range from about 1% to about 99% of the active ingredient, preferably from about 5% to about 70%, most preferably from about 10% to about 30%, by percent.

Methods of preparing these formulations or compositions include the step of bringing into association a compound of the present invention with a carrier and, optionally, one or more accessory ingredients. Typically, the formulation is prepared by: the compound of the present invention is uniformly and thoroughly mixed with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, the product is molded.

The compounds of formula I and their pharmaceutically acceptable salts and esters can be processed with pharmaceutically inert, inorganic or organic adjuvants to produce tablets, coated tablets, dragees, and hard gelatin capsules. Lactose, polyvinylpyrrolidone, hydroxypropylmethylcellulose, hydroxypropyl-cellulose, microcrystalline cellulose, corn starch or derivatives thereof, talc, stearic acid or its salts and the like can be used as excipients for, for example, tablets, troches and hard gelatin capsules.

Suitable excipients for soft gelatine capsules are, for example, vegetable oils, waxes, fats, semi-solid substances, liquid polyols and the like. Suitable adjuvants for producing solutions and syrups are, for example, H₂O, polyols, sucrose, invert sugar, glucose, and the like. Suitable adjuvants for injectable solutions are, for example, H₂O, alcohol, polyhydric alcohol, glycerin, vegetable oil and the like. Suitable excipients for suppositories are, for example, natural or hardened oils, waxes, fats, semi-solid or liquid polyols and the like. Suitable adjuvants for topical preparations are glycerides, semi-synthetic and synthetic glycerides, hydrogenated oils, liquid waxes, liquid paraffins, liquid fatty alcohols, sterols, polyethylene glycols and cellulose derivatives.

In addition, the pharmaceutical preparations can contain preservatives, solubilizers, viscosity-increasing substances, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavorants, salts for varying the osmotic pressure, buffers, masking agents or antioxidants. They may also contain other therapeutic substances.

The compounds of the present invention (compounds of formula I) may be prepared using the general reaction scheme as set forth below in scheme 1.

Scheme 1

The amino group in the compound of formula 2 may be protected with a suitable protecting group PG1 to give the compound of formula 3. The compound of formula 3 may be reacted with a chlorinating agent (e.g., POCl)₃) The protecting group PG1 in the compound of formula 5 can be removed to obtain the compound of formula 6 can be treated with an appropriately protected α -amino acid of formula 7 under dehydration conditions to obtain the compound of formula 8 the protecting group PG2 in the compound of formula 8 can be removed to obtain the compound of formula 9 can be treated with an appropriately protected α -amino acid of formula 10 under dehydration conditions to obtain the compound of formula 11 the protecting group PG3 in the compound of formula 11 can be removed to obtain the compound of formula I.

Methods of performing the above reactions and processes will be apparent to those skilled in the art based on the present disclosure, or may be derived in analogy to the examples. The starting materials are either commercially available or can be prepared by methods analogous to those described in the examples below.

Crystal form

When the compounds of the present invention are solids, it will be understood by those skilled in the art that these compounds and their salts may exist in various crystalline or polymorphic forms, all of which are intended to fall within the scope of the present invention and specific chemical formulas.

Examples

The compounds of the invention can be synthesized according to known techniques. The following examples and references are provided to aid in the understanding of the present invention. These examples are not intended, however, to limit the invention, the true scope of which is set forth in the appended claims. The nomenclature of the final products in the examples was generated using the functions available from AutoNom 2000Add-in v4.0SP2(ISIS Draw, function in Elsevier/MDL), AutoNom 2000TT v4.01.305(Elsevier/MDL), ChemDraw Pro Control 11.0.2(Cambridge Soft Corp.), or the Struct Name function of the electronic laboratory notes.

Example 1

(S) -2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride

Step 1: to a solution of 2- (tert-butoxycarbonyl) isoindoline-1-carboxylic acid (500mg,1.9mmol, Eq:1.00) and 2, 6-difluoroaniline (294mg,2.28mmol, Eq:1.2) in pyridine (10mL) at 0 deg.C was added POCl₃(437mg, 266. mu.L, 2.85mmol, Eq: 1.5). The reaction mixture was warmed to room temperature and stirred for 2 hours. The mixture was evaporated and water was added and the resulting mixture was extracted with EtOAc. The combined organics were washed with water, brine, MgSO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give 1- (2, 6-difluoro-phenylcarbamoyl) -1, 3-dihydro-isoindole-2-carboxylic acid tert-butyl ester (462mg) as a pale yellow foam.

Step 2: to 1- (2, 6-difluoro-phenylcarbamoyl) -1, 3-dihydro-isoindole-2-tert-butyl formate (460mg,1.23mmol, Eq:1.00) in CH₂Cl₂To a solution in (6mL) was added TFA (2mL) dropwise. The reaction mixture was stirred at room temperature for 1 hour. The solvent was evaporated to give 2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide trifluoroacetate salt, which was used without further purification.

And step 3: to a solution of 2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide trifluoroacetate (477mg,1.23mmol, Eq:1.00), (S) -2- (tert-butoxycarbonylamino) -3-methylbutyric acid (267mg,1.23mmol, Eq:1.00), and HATU (514mg,1.35mmol, Eq:1.1) in DMF (1.85mL) at 0 deg.C was added DIEA (476mg, 644. mu.L, 3.69mmol, Eq: 3). The reaction mixture was stirred at room temperature for 1 hour. Water was added and the mixture was extracted with EtOAc. The combined organics were washed with water, brine, MgSO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give two diastereomers. The less polar diastereomer was named { (S) -1- [ (S) -1- (2, 6-difluoro-phenylcarbamoyl) -1, 3-dihydro-isoindole-2-carbonyl]-2-methyl-propyl } -carbamic acid tert-butyl ester (220mg), which was isolated as a white foam.

And 4, step 4: to { (S) -1- [ (S) -1- (2, 6-difluoro-phenylcarbamoyl) -1, 3-dihydro-isoindole-2-carbonyl]-2-methyl-propyl } -carbamic acid tert-butyl ester (210mg, 443. mu. mol, Eq:1.00) in CH₂Cl₂To a solution in (3mL) was added TFA (1mL) dropwise. The reaction mixture was stirred at room temperature for 1 hour. The solution was evaporated to give (S) -2- ((S) -2-amino-3-methyl-butyryl) -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide trifluoroacetate salt, which was used without further purification.

And 5: to a solution of (S) -2- ((S) -2-amino-3-methyl-butyryl) -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide trifluoroacetate (216mg, 443. mu. mol, Eq:1.00), (S) -2- (tert-butoxycarbonyl-methyl-amino) -propionic acid (83.8mg, 443. mu. mol, Eq:1.00), HATU (185mg, 487. mu. mol, Eq:1.1) in DMF (500. mu.L) at 0 ℃ was added DIEA (172mg, 232. mu.L, 1.33mmol, Eq: 3). The reaction mixture was stirred at room temperature for 1 hour, diluted with water and usedAnd (4) extracting the EtOAc. The combined organics were washed with water, brine, MgSO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give ((S) -1- { (S) -1- [ (S) -1- (2, 6-difluoro-phenylcarbamoyl) -1, 3-dihydro-isoindole-2-carbonyl]-2-methyl-propylcarbamoyl } -ethyl) -methyl-carbamic acid tert-butyl ester (220mg) as a white foam.

Step 6: a solution of acetyl chloride (552mg, 500. mu.L, 7.03mmol, Eq:18.0) in MeOH (2mL) was added to a vial containing ((S) -1- { (S) -1- [ (S) -1- (2, 6-difluoro-phenylcarbamoyl) -1, 3-dihydro-isoindole-2-carbonyl ] -2-methyl-propylcarbamoyl } -ethyl) -methyl-carbamic acid tert-butyl ester (218mg, 390. mu. mol, Eq: 1.00). The mixture was stirred at room temperature for 1 hour. The mixture was evaporated and the resulting solid was dissolved in MeCN (3mL) and water (1mL) and the solution was lyophilized to give (S) -2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride (175mg) as a white powder with M/z ═ 459(M + H).

Example 2

(S) -2- [ (S) -2-cyclohexyl-2- ((S) -2-methylamino-propionylamino) -acetyl ] -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride

Step 1: to a solution of (S) -2- (tert-butoxycarbonyl) isoindoline-1-carboxylic acid (300mg,1.14mmol, Eq:1.00) and 2, 6-difluoroaniline (177mg,1.37mmol, Eq:1.2) in pyridine (3.00mL) at 0 deg.C was added POCl₃(262mg, 159. mu.L, 1.71mmol, Eq: 1.5). The mixture was warmed to room temperature and stirred for 2 hours. The mixture was evaporated, water was added and the mixture was extracted with EtOAc. The combined organics were washed with water, brine, MgSO₄Dried and concentrated in vacuo. Purifying the crude material by flash chromatography to obtainTo (S) -1- (2, 6-difluoro-phenylcarbamoyl) -1, 3-dihydro-isoindole-2-carboxylic acid tert-butyl ester (298mg), which was a pale yellow foam.

Step 2: to (S) -1- (2, 6-difluoro-phenylcarbamoyl) -1, 3-dihydro-isoindole-2-carboxylic acid tert-butyl ester in CH₂Cl₂To a solution in (3mL) was added TFA (1mL) dropwise. The reaction mixture was stirred at room temperature for 1 hour and the mixture was evaporated to give (S) -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide, which was used without further purification.

And step 3: DIEA (160mg, 216. mu.L, 1.24mmol, Eq:3) was added to a solution of (S) -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide (160mg, 412. mu. mol, Eq:1.00), (S) -2- (tert-butoxycarbonylamino) -2-cyclohexylacetic acid (106mg, 412. mu. mol, Eq:1.00), and HATU (172mg, 453. mu. mol, Eq:1.1) in DMF (500. mu.L) at 0 ℃. The reaction mixture was stirred at rt for 1h, diluted with water and extracted with EtOAc. The combined organics were washed with water, brine, MgSO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give { (S) -1-cyclohexyl-2- [ (S) -1- (2, 6-difluoro-phenyl-carbamoyl) -1, 3-dihydro-isoindol-2-yl]-2-oxo-ethyl } -carbamic acid tert-butyl ester (174mg) as a white foam.

And 4, step 4: to { (S) -1-cyclohexyl-2- [ (S) -1- (2, 6-difluoro-phenylcarbamoyl) -1, 3-dihydro-isoindol-2-yl]-2-oxo-ethyl } -carbamic acid tert-butyl ester (174mg, 339. mu. mol, Eq:1.00) in CH₂Cl₂To a solution in (3mL) was added TFA (1mL) dropwise. The reaction mixture was stirred at room temperature for 1 hour. The mixture was evaporated to give (S) -2- ((S) -2-amino-2-cyclohexyl-acetyl) -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide trifluoroacetate salt, which was used without further purification.

And 5: to (S) -2- ((S) -2-amino-2-cyclohexyl-acetyl) -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide trifluoroacetate (179mg, 339. mu. mol, Eq:1.00), (S) -2- (tert-butoxycarbonyl-methyl-amino) -propionic acid (64.2mg, 339. mu. mol, Eq:1.00), HATU: (R) ((R))142mg, 373. mu. mol, Eq:1.1) in DMF (1mL) DIEA (132mg, 178. mu.L, 1.02mmol, Eq:3) was added at 0 ℃. The reaction mixture was stirred at rt for 1h, diluted with water and extracted with EtOAc. The combined organics were washed with water, brine, MgSO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give ((S) -1- { (S) -1-cyclohexyl-2- [ (S) -1- (2, 6-difluoro-phenylcarbamoyl) -1, 3-dihydro-isoindol-2-yl)]-2-oxo-ethylcarbamoyl } -ethyl) -methyl-carbamic acid tert-butyl ester (174mg) as a white foam.

Step 6: a solution of acetyl chloride (552mg, 500. mu.L, 7.03mmol, Eq:24.2) in MeOH (2mL) was added to a vial containing ((S) -1- { (S) -1-cyclohexyl-2- [ (S) -1- (2, 6-difluoro-phenylcarbamoyl) -1, 3-dihydro-isoindol-2-yl ] -2-oxo-ethylcarbamoyl } -ethyl) -methyl-carbamic acid tert-butyl ester (174mg, 291. mu. mol, Eq: 1.00). The mixture was stirred at room temperature for 1 hour. The solvent was evaporated and the resulting solid was redissolved in MeCN (3mL) and water (1 mL). This was lyophilized to give (S) -2- [ (S) -2-cyclohexyl-2- ((S) -2-methylamino-propionylamino) -acetyl ] -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride (148mg) as a white powder with M/z of 499(M + H).

Example 3

(S) -2- [ (S) -2-cyclohexyl-2- ((S) -2-methylamino-propionylamino) -acetyl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride

Step 1: to a solution of (S) -3, 4-dihydro-1H-isoquinoline-1, 2-dicarboxylic acid 2-tert-butyl ester (500mg,1.8mmol, Eq:1.00) and 2, 6-difluoroaniline (279mg,2.16mmol, Eq:1.2) in pyridine (10.0mL) at 0 deg.C was added POCl₃(415mg, 252. mu.L, 2.7mmol, Eq: 1.5). The mixture was warmed to room temperature and stirred for 2 hours. Will dissolveThe solvent was evaporated, water was added and the mixture was extracted with EtOAc. The combined organics were washed with water, brine, MgSO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give (S) -tert-butyl 1- (2, 6-difluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinoline-2-carboxylate (380mg) as a white foam.

Step 2: to (S) -1- (2, 6-difluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester (50mg, 129. mu. mol, Eq:1.00) in CH₂Cl₂To a solution in (6mL) was added TFA (2mL) dropwise. The reaction mixture was stirred at room temperature for 1 hour. The mixture was evaporated to give (S) -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide trifluoroacetate salt, which was used without further purification.

And step 3: DIEA (50.1mg, 67.7. mu.L, 388. mu. mol, Eq:3) was added to a solution of (S) -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide trifluoroacetate (52mg, 129. mu. mol, Eq:1.00), (S) -2- (tert-butoxycarbonylamino) -2-cyclohexylacetic acid (33.3mg, 129. mu. mol, Eq:1.00), and HATU (54.1mg, 142. mu. mol, Eq:1.1) in DMF (300. mu.L) at 0 ℃. The reaction mixture was stirred at rt for 1h, diluted with water and extracted with EtOAc. The combined organics were washed with water, brine, MgSO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give { (S) -1-cyclohexyl-2- [ (S) -1- (2, 6-difluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinolin-2-yl]-2-oxo-ethyl } -carbamic acid tert-butyl ester (62mg) as a white foam.

And 4, step 4: to { (S) -1-cyclohexyl-2- [ (S) -1- (2, 6-difluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinolin-2-yl]-2-oxo-ethyl } -carbamic acid tert-butyl ester (62mg, 118. mu. mol, Eq:1.00) in CH₂Cl₂To a solution in (3mL) was added TFA (1mL) dropwise. The reaction mixture was stirred at room temperature for 1 hour. The mixture was evaporated to give (S) -2- ((S) -2-amino-2-cyclohexyl-acetyl) -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide trifluoroacetate salt, which was used without further purification.

And 5: to a solution of (S) -2- ((S) -2-amino-2-cyclohexyl-acetyl) -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide trifluoroacetate (63.6mg, 117. mu. mol, Eq:1.00), (S) -2- (tert-butoxycarbonyl-methyl-amino) -propionic acid (22.2mg, 117. mu. mol, Eq:1.00), HATU (49.1mg, 129. mu. mol, Eq:1.1) in DMF (500. mu.L) was added DIEA (45.5mg, 61.5. mu.L, 352. mu. mol, Eq:3) at 0 ℃. The reaction mixture was stirred at rt for 1h, diluted with water and extracted with EtOAc. The combined organics were washed with water, brine, MgSO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give ((S) -1- { (S) -1-cyclohexyl-2- [ (S) -1- (2, 6-difluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinolin-2-yl) -2]-2-oxo-ethylcarbamoyl } -ethyl) -methyl-carbamic acid tert-butyl ester (58mg) as a white foam.

Step 6: a solution of acetyl chloride (552mg, 500. mu.L, 7.03mmol, Eq:74.3) in MeOH (2mL) was added to a vial containing ((S) -1- { (S) -1-cyclohexyl-2- [ (S) -1- (2, 6-difluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinolin-2-yl ] -2-oxo-ethylcarbamoyl } -ethyl) -methyl-carbamic acid tert-butyl ester (58mg, 94.7. mu. mol, Eq: 1.00). The mixture was stirred at room temperature for 1 hour. The solvent was evaporated and the resulting solid was dissolved in MeCN (3mL) and water (1mL) and the solution was lyophilized to give (S) -2- [ (S) -2-cyclohexyl-2- ((S) -2-methylamino-propionylamino) -acetyl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride (50mg) as a white powder (50mg) with M/z ═ 513(M + H).

Example 4

(S) -2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2-chloro-6-fluoro-phenyl) -amide hydrochloride

Step 1: at 0 ℃ towards (S)2-tert-butyl 3, 4-dihydro-1H-isoquinoline-1, 2-dicarboxylate (100mg, 361. mu. mol, Eq:1) and 2-chloro-6-fluoroaniline (63.0mg, 433. mu. mol, Eq:1.2) in pyridine (2.00mL) was added POCl₃(82.9mg, 50.4. mu.L, 541. mu. mol, Eq: 1.5). The reaction mixture was warmed to room temperature and stirred for 2 hours. The mixture was evaporated and water was added to the residue. The mixture was extracted with EtOAc and the combined organics were washed with water, brine and MgSO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give (S) -tert-butyl 1- (2-chloro-6-fluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinoline-2-carboxylate (79mg) as a white solid.

Step 2: to (S) -1- (2-chloro-6-fluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester (78mg, 193. mu. mol, Eq:1.00) in CH₂Cl₂To a solution in (6mL) was added TFA (2mL) dropwise. The reaction mixture was stirred at room temperature for 1 hour. The mixture was evaporated to give (S) -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2-chloro-6-fluoro-phenyl) -amide trifluoroacetate salt, which was used without further purification.

And step 3: DIEA (74.1mg, 100. mu.L, 573. mu. mol, Eq:3) was added to a solution of (S) -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2-chloro-6-fluoro-phenyl) -amide trifluoroacetate (80mg, 191. mu. mol, Eq:1.00), (S) -2- (tert-butoxycarbonylamino) -3-methylbutyric acid (41.5mg, 191. mu. mol, Eq:1.00), and HATU (79.9mg, 210. mu. mol, Eq:1.1) in DMF (500. mu.L) at 0 ℃. The reaction mixture was stirred at rt for 1h, diluted with water and extracted with EtOAc. The combined organics were washed with water, brine, MgSO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give { (S) -1- [ (S) -1- (2-chloro-6-fluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinoline-2-carbonyl]-2-methyl-propyl } -carbamic acid tert-butyl ester (47mg) as a white foam.

And 4, step 4: to { (S) -1- [ (S) -1- (2-chloro-6-fluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinoline-2-carbonyl]-2-methyl-propyl } -carbamic acid tert-butyl ester (47mg, 93.3. mu. mol, Eq:1.00) in CH₂Cl₂To a solution in (3mL) was added TFA (1mL) dropwise. Mixing the reaction mixtureStir at room temperature for 1 hour. The mixture was evaporated to give (S) -2- ((S) -2-amino-3-methyl-butyryl) -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2-chloro-6-fluoro-phenyl) -amide trifluoroacetate salt, which was used without further purification.

And 5: to a solution of (S) -2- ((S) -2-amino-3-methyl-butyryl) -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2-chloro-6-fluoro-phenyl) -amide trifluoroacetate (48mg, 92.7. mu. mol, Eq:1.00), (S) -2- (tert-butoxycarbonylamino) propionic acid (17.5mg, 92.7. mu. mol, Eq:1.00), HATU (38.8mg, 102. mu. mol, Eq:1.1) in DMF (300. mu.L) was added DIEA (35.9mg, 48.6. mu.L, 278. mol, Eq:3) at 0 ℃. The reaction mixture was stirred at rt for 1h, diluted with water and extracted with EtOAc. The combined organics were washed with water, brine, MgSO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give ((S) -1- { (S) -1- [ (S) -1- (2-chloro-6-fluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinoline-2-carbonyl]-2-methyl-propylcarbamoyl } -ethyl) -methyl-carbamic acid tert-butyl ester (36mg) as a white foam.

Step 6: a solution of acetyl chloride (552mg, 500. mu.L, 7.03mmol, Eq:115) in MeOH (2mL) was added to a vial containing ((S) -1- { (S) -1- [ (S) -1- (2-chloro-6-fluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinoline-2-carbonyl ] -2-methyl-propylcarbamoyl } -ethyl) -methyl-carbamic acid tert-butyl ester (36mg, 61.1. mu. mol, Eq: 1.00). The mixture was stirred at room temperature for 1 hour. The solvent was evaporated and the resulting solid was dissolved in MeCN (3mL) and water (1mL) and lyophilized to give (S) -2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2-chloro-6-fluoro-phenyl) -amide hydrochloride (30mg) as a white powder, M/z ═ 489(M + H).

Example 5

6-chloro-2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide

Step 1: in a 50mL round-bottom flask, 2- (tert-butoxycarbonyl) -6-chloro-1, 2,3, 4-tetrahydroisoquinoline-1-carboxylic acid (0.5g,1.6mmol, Eq:1.00), 2, 6-difluoroaniline (228mg, 190. mu.l, 1.76mmol, Eq:1.1), and TEA (487mg, 671. mu.l, 4.81mmol, Eq:3) were mixed with DCM (10mL) to give a colorless solution, and phosphoryl chloride (295mg, 179. mu.l, 1.92mmol, Eq:1.2) was added. After 2 days, the reaction mixture was diluted with DCM and washed with 1N HCl, water and brine. Subjecting the organic layer to Na₂SO₄Dried and concentrated in vacuo to give 0.6076g of tert-butyl 6-chloro-1- (2, 6-difluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinoline-2-carboxylate as a brown foam which is used without purification. 89.6% MS M/z322.9 (M-BOC).

Step 2: in a 50mL pear-shaped vial, tert-butyl 6-chloro-1- (2, 6-difluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinoline-2-carboxylate (0.6g,1.42mmol, Eq:1.00) is mixed with DCM (25mL) to give a light brown solution. TFA (4.85g,3.28mL,42.6mmol, Eq:30) was added. After 1h, the reaction mixture was concentrated, the residue was dissolved in EtOAc, and the mixture was washed with 1N NaOH and brine. Subjecting the organic layer to Na₂SO₄Dried and concentrated in vacuo to give 0.3937g of 6-chloro-1, 2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide as a brown oil, which is used without purification. 86.0% MS M/z 323.0(M + H).

And step 3: in a 100mL round bottom flask, BOC-VAL-OH (315mg,1.45mmol, Eq:1.2), 6-chloro-1, 2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide (0.39g,1.21mmol, Eq:1.00) and HATU (551mg,1.45mmol, Eq:1.2) were mixed with DMF (10mL) to give a brown solution and TEA (367mg, 505. mu.l, 3.63mmol, Eq:3) was added. After 3 hours, the reaction mixture was diluted with EtOAc and diluted with 1:1 saturated NaHCO₃Brine and brine wash. Subjecting the organic layer to Na₂SO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give 0.3579g of { (S) -1- [ 6-chloro-1- (2, 6-difluoro-phenylcarbamoyl)Yl) -3, 4-dihydro-1H-isoquinoline-2-carbonyl]-2-methyl-propyl } -carbamic acid tert-butyl ester, which is a light yellow foam. 56.7% MS M/z 522.0(M + H).

And 4, step 4: in a 100mL pear-shaped flask, (2S) -tert-butyl 1- (6-chloro-1- (2, 6-difluorophenylcarbamoyl) -3, 4-dihydroisoquinolin-2 (1H) -yl) -3-methyl-1-oxobutan-2-ylcarbamate (0.3579g, 686. mu. mol, Eq:1.00) was mixed with DCM (20mL) to give a pale yellow solution, and TFA (2.35g,1.58mL,20.6mmol, Eq:30) was added. After 1h, the reaction mixture was concentrated, the residue was dissolved in EtOAc, and the mixture was washed with 1N NaOH and brine. Subjecting the organic layer to Na₂SO₄Dried and concentrated in vacuo to give 0.2377g of 2- ((S) -2-amino-3-methyl-butyryl) -6-chloro-1, 2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide as a pale yellow foam, which is used without purification. 82.2% MS M/z 422.1(M + H).

And 5: in a 50mL flask, 2- ((S) -2-amino-3-methyl-butyryl) -6-chloro-1, 2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide (0.2377g, 563. mu. mol, Eq:1.00), BOC-N-ME-ALA-OH (137mg, 676. mu. mol, Eq:1.2), and HATU (257mg, 676. mu. mol, Eq:1.2) were mixed with DMF (6mL) to give a light yellow solution, and TEA (171mg, 236. mu.l, 1.69mmol, Eq:3) was added. After 2 hours, the reaction mixture was diluted with EtOAc and washed with brine. Subjecting the organic layer to Na₂SO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give 0.1976g ((S) -1- { (S) -1- [ 6-chloro-1- (2, 6-difluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinoline-2-carbonyl)]-2-methyl-propylcarbamoyl } -ethyl) -methyl-carbamic acid tert-butyl ester as white foam. 57.8% MS M/z607.1(M + H).

Step 6: in a 20mL vial, ((S) -1- { (S) -1- [ 6-chloro-1- (2, 6-difluoro-phenylcarbamoyl) -3, 4-dihydro-1H-isoquinoline-2-carbonyl]-2-methyl-propylcarbamoyl } -ethyl) -methyl-carbamic acid tert-butyl ester (0.1976g, 325. mu. mol, Eq:1.00) was mixed with DCM (6mL) to give a colorless solution and TFA (1.11g, 752. mu.l, 9.76mmol, Eq:30) was added. After 1 hour, the reaction mixture was concentrated and the residue was dissolved in EtOAIn c, wash with 1N NaOH and brine. Subjecting the organic layer to Na₂SO₄Dried and concentrated in vacuo to give 125.8mg of a diastereomeric mixture as a white foam. The diastereomers were separated by Supercritical Fluid Chromatography (SFC) (using a 220mm Diacel IA column, 25% EtOH,70 mL/min). Each diastereomer was isolated and separated from MeCN/H₂And (4) freeze-drying. The first compound eluted to give 50.2mg of a white solid. The second compound eluted to give 47.2mg of the title compound as a white solid, the structure of which was determined based on activity in the TR-FRET assay. 57.6% MS M/z 507.2(M + H).

Example 6

6-fluoro-2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide

Step 1: in a 50mL round-bottom flask, 6-fluoro-1, 2,3, 4-tetrahydroisoquinoline-1-carboxylic acid hydrochloride (500mg,2.16mmol, Eq:1.00) and 2N NaOH (3.24mL,6.48mmol, Eq:3) were mixed with t-BuOH (3.00mL) to give a light brown solution, and BOC-anhydride (565mg, 601. mu.l, 2.59mmol, Eq:1.2) was added. After stirring for 18 hours, the reaction mixture was diluted with water and extracted with diethyl ether. The aqueous layer was saturated with KHSO₄Acidified and extracted with EtOAc. The combined organic extracts were washed with brine, over Na₂SO₄Drying, filtration and concentration gave 0.6448g of 2-tert-butyl 6-fluoro-3, 4-dihydro-1H-isoquinoline-1, 2-dicarboxylate as a white foam which was used without purification. 100% MS M/z193.9 (M-BOC).

Step 2: to a solution of 2-tert-butyl 6-fluoro-3, 4-dihydro-1H-isoquinoline-1, 2-dicarboxylate (0.37g,1.25mmol, Eq:1.00) and 2, 6-difluoroaniline (194mg, 162. mu.l, 1.5mmol, Eq:1.2) in pyridine (7mL) was added phosphorus oxychloride (288mg, 175. mu.l, 1.88mmol, Eq:1.5) at 0 deg.C). The reaction mixture was warmed to room temperature, stirred for 18 h, diluted with water and extracted with EtOAc. The combined organics were washed with 1N HCl, water, brine, Na₂SO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give 0.2181g of tert-butyl 1- (2, 6-difluoro-phenylcarbamoyl) -6-fluoro-3, 4-dihydro-1H-isoquinoline-2-carboxylate as a white foam. 42.8% MS M/z 428.9(M + H).

And step 3: in a 50mL round bottom flask, tert-butyl 1- (2, 6-difluoro-phenylcarbamoyl) -6-fluoro-3, 4-dihydro-1H-isoquinoline-2-carboxylate (0.218g, 536. mu. mol, Eq:1.00) is mixed with DCM (7.00mL) to give a colorless solution and TFA (1.83g,1.24mL,16.1mmol, Eq:30) is added. After 1 hour, the mixture was concentrated to give 0.2867g of 6-fluoro-1, 2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide trifluoroacetate salt as a yellow oil which was used without purification. 127% MS M/z 307.1(M + H).

And 4, step 4: in a 50mL round bottom flask, BOC-VAL-OH (140mg, 643. mu. mol, Eq:1.2) and 6-fluoro-1, 2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide trifluoroacetate (225mg, 536. mu. mol, Eq:1.00) are mixed with DMF (3mL) to give a light yellow solution and TEA (217mg, 299. mu.l, 2.14mmol, Eq:4) is added. To this solution were added HBTU (244mg, 643. mu. mol, Eq:1.2) and HOBT. H₂A solution of O (98.5mg, 643. mu. mol, Eq:1.2) in DMF (3 mL). After 2 hours, the reaction mixture was diluted with EtOAc and diluted with 1:1 saturated NaHCO₃Brine, brine wash, and pass the organic layer over Na₂SO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give 97.8mg { (S) -1- [1- (2, 6-difluoro-phenylcarbamoyl) -6-fluoro-3, 4-dihydro-1H-isoquinoline-2-carbonyl]-2-methyl-propyl } -carbamic acid tert-butyl ester, which is a white foam. 36.1% MS M/z506.0(M + H).

And 5: in a 20mL vial, { (S) -1- [1- (2, 6-difluoro-phenylcarbamoyl) -6-fluoro-3, 4-dihydro-1H-isoquinoline-2-carbonyl ] -2-methyl-propyl } -carbamic acid tert-butyl ester (97.8mg, 193. mu. mol, Eq:1.00) was mixed with DCM (4mL) to give a colorless solution and TFA (662mg, 447. mu.l, 5.8mmol, Eq:30) was added. After 1 hour, the mixture was concentrated to give 0.1346g of 2- ((S) -2-amino-3-methyl-butyryl) -6-fluoro-1, 2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide trifluoroacetate salt as a light brown oil which was used without purification. 134% MS M/z406.1(M + H).

Step 6: in a 20mL vial, 2- ((S) -2-amino-3-methyl-butyryl) -6-fluoro-1, 2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide trifluoroacetate (100mg, 193. mu. mol, Eq:1.00), BOC-N-ME-ALA-OH (47.1mg, 232. mu. mol, Eq:1.2) and HATU (88.1mg, 232. mu. mol, Eq:1.2) were mixed with DMF (3mL) to give a light brown solution and TEA (97.6mg, 135. mu.L, 965. mu. mol, Eq:5) was added. After 2.5 h, the mixture was diluted with EtOAc and washed with brine. Subjecting the organic layer to Na₂SO₄Dried and concentrated in vacuo. The crude material was purified by flash chromatography to give 77.1mg of ((S) -1- { (S) -1- [1- (2, 6-difluoro-phenylcarbamoyl) -6-fluoro-3, 4-dihydro-1H-isoquinoline-2-carbonyl]-2-methyl-propylcarbamoyl } -ethyl) -methyl-carbamic acid tert-butyl ester as white solid. 67.6% MS M/z 591.1(M + H).

And 7: in a 20mL vial, ((S) -1- { (S) -1- [1- (2, 6-difluoro-phenylcarbamoyl) -6-fluoro-3, 4-dihydro-1H-isoquinoline-2-carbonyl]-2-methyl-propylcarbamoyl } -ethyl) -methyl-carbamic acid tert-butyl ester (77.1mg, 131. mu. mol, Eq:1.00) was mixed with DCM (3.00mL) to give a colorless solution and TFA (447mg, 302. mu.l, 3.92mmol, Eq:30) was added. After 30 min, the mixture was concentrated, the residue was dissolved in EtOAc and washed with 1N NaOH, water and Na₂SO₄Drying and concentration in vacuo gave 47.9mg of a mixture of diastereomers as a white foam. Each diastereomer was isolated and separated from MeCN/H₂And (4) freeze-drying. The first compound eluted to give 22.6mg of a white solid. The second compound elutes to give 13.7mg of the title compound as a white solid whose structure is determined based on activity in the TR-FRET assay. 21.4% MS M/z 491.3(M + H).

The compounds listed in table 1 below were prepared following the procedure described above for example 1, using the amines listed in the table in place of 2, 6-difluoroaniline in step 1.

TABLE 1

The compounds listed in table 2 below were prepared following the procedure described above for example 1, using the amines listed in the table instead of 2, 6-difluoroaniline in step 1, and omitting the separation of the diastereoisomers described in step 3.

TABLE 2

The compounds listed in Table 3 were prepared by following the procedure as described above for example 2, using the carboxylic acids listed in the Table in place of (S) -2- (tert-butoxycarbonylamino) -2-cyclohexylacetic acid in step 3.

TABLE 3

The compounds listed in Table 4 below were prepared by following the procedure as described above for example 3, using the carboxylic acids listed in the table in place of (S) -2- (tert-butoxycarbonylamino) -2-cyclohexylacetic acid in step 3.

TABLE 4

The compounds listed in Table 5 below were prepared following the procedure described above for example 4, using the amines listed in the table in place of 2-chloro-6-fluoroaniline in step 1.

TABLE 5

The compounds listed in Table 6 below were prepared according to the procedure described in example 5 above, using 1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid listed in the table instead of 2- (tert-butoxycarbonyl) -6-chloro-1, 2,3, 4-tetrahydroisoquinoline-1-carboxylic acid in step 1.

TABLE 6

The compounds listed in Table 7 below were prepared according to the procedure described in example 6 above, using 1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid listed in the table instead of 2- (tert-butoxycarbonyl) -6-chloro-1, 2,3, 4-tetrahydroisoquinoline-1-carboxylic acid in step 1.

TABLE 7

Example 31

(S) -2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -1,2,3, 4-tetrahydro-isoquinoline-3-carboxylic acid (2, 6-difluoro-phenyl) -amide

Step 1: to a solution of (S) -2- (tert-butoxycarbonyl) -1,2,3, 4-tetrahydroisoquinoline-3-carboxylic acid (200mg, 721. mu. mol, Eq:1.00) and 2, 6-difluoroaniline (121mg, 101. mu.L, 938. mu. mol, Eq:1.3) in pyridine (2mL) was added phosphorus oxychloride (166mg, 101. mu.L, 1.08mmol, Eq:1.5) dropwise at 0 ℃. The cooling bath was removed and the reaction mixture was stirred at room temperature overnight. The reaction mixture was concentrated in vacuo and water was added to the residue. The resulting mixture was extracted with EtOAc, and the combined organic layers were washed with 0.1M HCl, 0.1M NaOH, and brine, and concentrated in vacuo. The crude material was purified by flash chromatography to give (S) -3- (2, 6-difluorophenylcarbamoyl) -3, 4-dihydroisoquinoline-2 (1H) -carboxylic acid tert-butyl ester as a colorless oil (90mg), M/z ═ 411(M + Na).

Step 2: to a solution of tert-butyl (S) -3- (2, 6-difluorophenylcarbamoyl) -3, 4-dihydroisoquinoline-2 (1H) -carboxylate (90mg, 232. mu. mol, Eq:1.00) in DCM (2mL) was added TFA (2mL,26.0mmol, Eq:112), and the resulting solution was stirred at room temperature for 1 hour. The reaction mixture was concentrated in vacuo, and the residue was azeotroped with N-heptane to give (S) -N- (2, 6-difluorophenyl) -1,2,3, 4-tetrahydroisoquinoline-3-carboxamide trifluoroacetate salt, which was used without purification (93mg), M/z 289(M + H).

And step 3: to a solution of (S) -2- (tert-butoxycarbonylamino) -3-methylbutyric acid (60mg, 277. mu. mol, Eq:1.2) and DIEA (120. mu.L, 693. mu. mol, Eq:3) in DMF (1mL) was added HATU (105mg, 277. mu. mol, Eq:1.2), and the resulting solution was stirred at room temperature for 15 minutes. Then a solution of (S) -N- (2, 6-difluorophenyl) -1,2,3, 4-tetrahydroisoquinoline-3-carboxamide trifluoroacetate salt (93mg, 231. mu. mol, Eq:1.00) in DMF (1mL) was added and stirring continued at room temperature. After 1 hour, the reaction mixture was diluted with EtOAc and washed with 0.1M NaOH, 0.1M HCl and brine. The organic layer was concentrated in vacuo and the residue was azeotroped with n-heptane to give tert-butyl (S) -1- ((S) -3- (2, 6-difluorophenylcarbamoyl) -3, 4-dihydroisoquinolin-2 (1H) -yl) -3-methyl-1-oxobutan-2-ylcarbamate as a white solid which was used without purification (130mg), M/z ═ 488(M + H).

And 4, step 4: to a solution of tert-butyl (S) -1- ((S) -3- (2, 6-difluorophenylcarbamoyl) -3, 4-dihydroisoquinolin-2 (1H) -yl) -3-methyl-1-oxobutan-2-ylcarbamate (0.11g, 226. mu. mol, Eq:1.00) in DCM (2mL) was added TFA (2.96g,2mL,26.0mmol, Eq:115), and the resulting solution was stirred at room temperature for 1 hour. The reaction mixture was concentrated in vacuo, and the residue was azeotroped with N-heptane to give (S) -2- ((S) -2-amino-3-methylbutyryl) -N- (2, 6-difluorophenyl) -1,2,3, 4-tetrahydroisoquinoline-3-carboxamide trifluoroacetate salt, which was used without purification (0.11 g).

And 5: to a solution of (S) -2- (tert-butoxycarbonyl (methyl) amino) propionic acid (53.5mg, 263. mu. mol, Eq:1.2) and DIEA (190. mu.L, 1.1mmol, Eq:5) in DMF (2mL) was added HATU (100mg, 263. mu. mol, Eq: 1.2). The mixture was stirred for 20 minutes, and a solution of (S) -2- ((S) -2-amino-3-methylbutyryl) -N- (2, 6-difluorophenyl) -1,2,3, 4-tetrahydroisoquinoline-3-carboxamide trifluoroacetate (0.11g, 219. mu. mol, Eq:1.00) in DMF (1mL) was added, and the resulting solution was stirred at room temperature overnight.

The reaction mixture was diluted with EtOAc, washed with 0.1M HCl, 0.1M NaOH, and brine. The organic layer was concentrated in vacuo and the residue was azeotroped with n-heptane. The resulting material was purified by flash chromatography to give tert-butyl (S) -1- ((S) -3- (2, 6-difluorophenylcarbamoyl) -3, 4-dihydroisoquinolin-2 (1H) -yl) -3-methyl-1-oxobutan-2-ylamino) -1-oxoprop-2-yl (methyl) carbamate as a colorless oil (0.14g), M/z ═ 573(M + H).

Step 6: to a solution of tert-butyl (S) -1- ((S) -1- ((S) -3- (2, 6-difluorophenylcarbamoyl) -3, 4-dihydroisoquinolin-2 (1H) -yl) -3-methyl-1-oxobutan-2-ylamino) -1-oxoprop-2-yl (methyl) carbamate (138mg, 241. mu. mol, Eq:1.00) in DCM (2mL) was added TFA (2.96g,2mL,26.0mmol, Eq:108), and the resulting solution was stirred at room temperature for 1 hour. The mixture was concentrated in vacuo and saturated NaHCO₃To the residue was added and the resulting mixture was extracted with DCM. Subjecting the combined extracts to Na₂SO₄Dried and concentrated in vacuo. Treating the residue with diethyl ether/hexane to obtain (S) -2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl]-1,2,3, 4-tetrahydro-isoquinoline-3-carboxylic acid (2, 6-difluoro-phenyl) -amide as a white solid which was used without purification (70mg), M/z ═ 473(M + H).

Example 48

Biochemical assay

TR-FRET assays for BIR2 and BIR3

The ability of the test compound to inhibit binding of BIR2 and/or BIR3 domain of XIAP protein to peptide a (SMAC-derived peptides as described below) indicates that the test compound acts as a SMAC-mimetic, resulting in reactivation of the apoptotic pathway of the cell.

The peptide AVPIAQKSEK- (-biotin) -OH 1:2TFA ("peptide a") was identified as a substrate for the TR-FRET assay by screening the BIR2 domain and BIR3 domain of 6x histidine-tagged XIAP against a panel of 29 peptides synthesized from the sequences reported by Sweeny et al (Biochemistry,2006,45, 1474014748). The peptides were labeled with the fluorescent label FITC or TAMRA, and Kd values were determined by fluorescence polarization assay. The determination of the sequence AVPIAQKSEK is most suitable for use in assays. The peptide sequence was derivatized with biotin to provide AVPIAQKSEK- (-biotin) -OH 1:2TFA as a substrate for TR-FRET assays.

XIAP protein sequences were obtained from the SWISS-PROT protein sequence database, and BIR2 and BIR3 domains were derived therefrom. The sequence of the BIR2 domain used in the TR-FRET assay was MRHHHHHHRDHFALDRPSETHADYLLRTGQVVDISDTIYPRNPAMYSEEARLKSFQNWPDYAHLTPRELASAGLYYTGIGDQVQCFACGGKLKNWEPGDRAWSEHRRHFPNCFFVLGRNLNIRSE.

The BIR3 domain used in the TR-FRET assay has a sequence of MRHHHHHHRSDAVSSDRNFPNSTNLPRNPSMADYEARIFTFGTWIYSVNK

EQLARAGFYALGEGDKVKCFHCGGGLTDWKPSEDPWEQHAKWYPGCKYLL

EQKGQEYINNIHLTHSLEECLVRTT.

10nM of the 6 × histidine-tagged BIR2 domain (corresponding to amino acid 124. sup. 240 of XIAP), or BIR3 domain (corresponding to amino acid 241. sup. 356 of XIAP) was mixed with the 20nM peptide AVPIAQKSEK- (-biotin) -OH 1:2TFA in the presence of 50mM Tris-Cl, pH 7.5, 100mM NaCl, 1mM Dithiothreitol (DTT) and 0.1mg/mL Bovine Serum Albumin (BSA). After incubation at 37 ℃ for 45 min, europium-streptavidin and allophycocyanin-binding anti-histidine antibodies were added to final concentrations of 1.5nM and 15nM, respectively. The time resolved fluorescence resonance energy transfer (TR-FRET) signal was measured after 1 hour at room temperature. Test compound potency was assessed at 10-fold serial dilution concentrations. The percent inhibition at each concentration was determined to yield the IC of each test compound₅₀The value is obtained.

These values are listed in table 8 below.

TABLE 8

Claims

1. A process for the preparation of a compound having a structure,

the compound is selected from:

(S) -N- (benzo [ b ] thiophen-4-yl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxylic acid amide;

(S) -N- (2, 6-difluorophenyl) -2- ((S) -3, 3-dimethyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxylic acid amide;

2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) -N-phenylisoindoline-1-carboxamide;

n- (2-methoxyphenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide;

n-benzyl-2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide;

2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) -N-phenethylisoindoline-1-carboxamide;

(S) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) -N- (naphthalen-1-yl) isoindoline-1-carboxylic acid amide;

(S) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) -N- (naphthalen-2-yl) isoindoline-1-carboxylic acid amide;

(S) -N- (2-fluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide;

(S) -N- (2-chlorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide;

(S) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) -N- (5,6,7, 8-tetrahydronaphthalen-1-yl) isoindoline-1-carboxamide;

n- (isoquinolin-1-yl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide;

(S) -N- (1-methyl-1H-indol-4-yl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide;

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -3-methoxy-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxylic acid amide;

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3R) -3-hydroxy-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxylic acid amide;

(S) -N- (2, 6-difluorophenyl) -2- ((S) -4-methyl-2- ((S) -2- (methylamino) propionylamino) pentanoyl) isoindoline-1-carboxamide;

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -2- ((S) -2- (methylamino) propionylamino) -3-phenylbutyryl) isoindoline-1-carboxylic acid amide;

(S) -N- (2-fluoro-6-methylphenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxylic acid amide;

(S) -N- (2, 6-difluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide;

(S) -N- (2, 6-dichlorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide;

(S) -N- (2, 6-difluorophenyl) -2- ((S) -3, 3-dimethyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide;

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3R) -3-methoxy-2- ((S) -2- (methylamino) propionylamino) butanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide;

(S) -2- [ (S) -2-cyclohexyl-2- ((S) -2-methylamino-propionylamino) -acetyl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide;

(S) -2- ((S) -2-cyclopentyl-2- ((S) -2- (methylamino) propionylamino) acetyl) -N- (2, 6-difluorophenyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide;

(S) -N- (2, 6-difluorophenyl) -2- ((S) -2- (methylamino) propionylamino) -2-phenylacetyl) isoindoline-1-carboxamide;

(S) -2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide;

((S) -2- [ (S) -2-cyclohexyl-2- ((S) -2-methylamino-propionylamino) -acetyl ] -2, 3-dihydro-1H-isoindole-1-carboxylic acid (2, 6-difluoro-phenyl) -amide;

(S) -N- (2, 6-difluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxamide;

(S) -2- [ (S) -3-methyl-2- ((S) -2-methylamino-propionylamino) -butyryl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2-chloro-6-fluoro-phenyl) -amide;

(S) -N- (2, 6-dichlorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butanoyl) isoindoline-1-carboxamide;

(S) -N- (2-chloro-6-fluorophenyl) -2- ((S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) butyryl) isoindoline-1-carboxylic acid amide;

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) pentanoyl) isoindoline-1-carboxylic acid amide;

(S) -N- (2, 6-difluorophenyl) -2- ((S) -2- (methylamino) propionylamino) -2- (tetrahydro-2H-pyran-4-yl) acetyl) isoindoline-1-carboxylic acid amide;

(S) -2- ((S) -2-cyclopentyl-2- ((S) -2- (methylamino) propionylamino) acetyl) -N- (2, 6-difluorophenyl) isoindoline-1-carboxylic acid amide;

(S) -N- (2, 6-difluorophenyl) -2- ((S) -2- (methylamino) propionylamino) -2- (tetrahydro-2H-pyran-4-yl) acetyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide; and

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) pentanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide;

or a pharmaceutically acceptable salt of each of the foregoing compounds.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the pharmaceutically acceptable salt is selected from the group consisting of:

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -2- ((S) -2- (methylamino) propionylamino) -3-phenylbutyryl) isoindoline-1-carboxamide hydrochloride;

(S) -2- [ (S) -2-cyclohexyl-2- ((S) -2-methylamino-propionylamino) -acetyl ] -1,2,3, 4-tetrahydro-isoquinoline-1-carboxylic acid (2, 6-difluoro-phenyl) -amide hydrochloride;

(S) -N- (2, 6-difluorophenyl) -2- ((S) -2- (methylamino) propionylamino) -2- (tetrahydro-2H-pyran-4-yl) acetyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride; and

(S) -N- (2, 6-difluorophenyl) -2- ((2S,3S) -3-methyl-2- ((S) -2- (methylamino) propionylamino) pentanoyl) -1,2,3, 4-tetrahydroisoquinoline-1-carboxamide hydrochloride.

3. A pharmaceutical composition comprising a compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient, and a pharmaceutically acceptable carrier or excipient.

4. Use of a compound of claim 1, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the therapeutic and/or prophylactic treatment of cancer mediated by BIR2 and/or BIR 3.