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NZ726105B2 - Dipicolylamine derivatives and their pharmaceutical uses - Google Patents

Dipicolylamine derivatives and their pharmaceutical uses Download PDF

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Publication number
NZ726105B2
NZ726105B2 NZ726105A NZ72610515A NZ726105B2 NZ 726105 B2 NZ726105 B2 NZ 726105B2 NZ 726105 A NZ726105 A NZ 726105A NZ 72610515 A NZ72610515 A NZ 72610515A NZ 726105 B2 NZ726105 B2 NZ 726105B2
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New Zealand
Prior art keywords
radical
monovalent
compound
bivalent
aryl
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NZ726105A
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NZ726105A (en
Inventor
Yusheng Chao
Chiungtong Chen
Kakshan Shia
Lunkelvin Tsou
Chienhuang Wu
Yu Sheng Chao
Chiung Tong Chen
Kak Shan Shia
Lun Kelvin Tsou
Chien Huang Wu
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Molecular Targeting Technologies Inc
National Health Research Institutes
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Priority claimed from PCT/US2015/031386 external-priority patent/WO2015179299A1/en
Publication of NZ726105A publication Critical patent/NZ726105A/en
Publication of NZ726105B2 publication Critical patent/NZ726105B2/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/436—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having oxygen as a ring hetero atom, e.g. rapamycin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/444—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring heteroatom, e.g. amrinone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D305/00—Heterocyclic compounds containing four-membered rings having one oxygen atom as the only ring hetero atoms
    • C07D305/14—Heterocyclic compounds containing four-membered rings having one oxygen atom as the only ring hetero atoms condensed with carbocyclic rings or ring systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
    • C07D491/14—Ortho-condensed systems
    • C07D491/147—Ortho-condensed systems the condensed system containing one ring with oxygen as ring hetero atom and two rings with nitrogen as ring hetero atom
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/22—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains four or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains three hetero rings
    • C07D498/18—Bridged systems

Abstract

Dipicolylamine compounds of Formula (I) set forth herein, that exhibit binding to phosphatidylserine. Also disclosed are pharmaceutical compositions containing metal ions and these compounds. Further disclosed is a method for treating a condition associated with cells containing inside-out phosphatidylserine, such as cancer, with these compounds dylserine, such as cancer, with these compounds

Description

WO 79299 DIPICOLYLAMINE DERIVATIVES AND THEIR PHARMACEUTICAL USES CROSS REFERENCE TO RELATED APPLICATIONS This application claims the t of US Provisional Application No. ,725, filed on May 22, 2014, the content of which is hereby incorporated by nce in its entirety BACKGROUND Phosphatidylserine (PS), a membrane phospholipid, is typically zed to the internal surface of the membrane of a healthy cell. Under certain circumstances, PS is also found on the external surface. See Leventis et al., Annu. Rev. s. 2010, 39, 407-27.
More specifically, PS is exposed on the external surface of cancer cells. See Thorpe et al., Breast Cancer Res Treat 1995, 36(2), 237-51; Ran et al., Int. J. Radiat. Oncol. Biol.
Phys. 2002, 54(5), 1479-84; and Thorpe, Thromb. Res. 2010, 125 Suppl 2, S134-137.
Further, recent studies found that PS exists in tumor vasculatures and tumor-derived microvessels. See Stafford et al., Neoplasz'a. 2011, 13, 299-308; and Yin et al., Cancer Immunology Research 2013, 1, 1-13. Moreover, in many enic particles such as bacteria and viruses, PS is exposed at high levels on the external surface. See Huang et al., Cancer Res. 2005, 65(10), 4408-16; and White et al., Bioconjug. Chem. 2010, 21(7), 1297- 1304. Finally, PS has been found on the ourter e of cells in which cell death pathways have been dysregulated. For example, in addition to cancer, conditions such as neurodegenerative disorders, cardiovascular disease, autoimmune diseases, and metabolic disorders demonstrate e localization of PS. See Smith et al., Mol. Pharm. 2011, 8(2), 583-90. Thus, PS provides a valuable target for delivery of therapeutic agents for treating the conditions mentioned above.
Protein Annexin V is currently used to deliver therapeutic agents via binding to PS.
However, this binding requires high levels of Ca2+, which might activate “scramblases” that could extemalize PS in nearby normal cells, resulting in red targeting of the normal cells.
There is a need to develop a delivery agent that selectively associates with disease- relevant PS to achieve site-specific ry of a therapeutic agent.
SUMMARY This invention is based on an cted discovery that n lylamine derivatives are effective in delivery a therapeutic agent to a target disease site that has phosphatidylserine on the external surfaces of cell membranes.
In one aspect, this invention relates to compounds of formula (1) shown below: B1 82 Z x/ \L1/ \L2/ In this formula, each of A1, A2, A3, A4, A5, A6, and B1, independently, is a C1-C6 bivalent tic radical, a C1-C6 bivalent heteroaliphatic radical, a bivalent aryl l, or a nt heteroaryl radical; B2 is a bond, a C1-C6 nt aliphatic radical, a C1-C6 bivalent heteroaliphatic radical, a bivalent aryl radical, a bivalent heteroaryl radical, Dl-NRl-C(O)-D2, D1-C(O)NR1-D2-NR1’-C(O)-D3, D1-D2-C(O)-NR1-C(O)-D3, or D1-D2-D3, each of D1, D2, D3, independently, being a C1-C6 bivalent aliphatic radical, a C1-C6 bivalent heteroaliphatic radical, a bivalent aryl radical, a bivalent aryl radical, a C1-C10 bivalent aralkyl radical, or a C1-C10 bivalent heteroaralkyl radical, and each of R1 and R1’, independently, being H, a C1-C6 bivalent heteroaliphatic radical, a bivalent aryl l, a bivalent heteroaryl radical, or a C1-C10 bivalent aralkyl l; each of L1 and L2, independently, is a bond, NR2, NR2C(O), NR2C(S), NR2CR3R4, NR2SO2, NR2C(O)NR3 or )NR3, each of R2, R3, and R4, independently, being H, a C1-C6 monovalent aliphatic radical, a C1-C6 monovalent heteroaliphatic radical, a monovalent aryl radical, a monovalent heteroaryl radical, a C1-C14 monovalent aralkyl radical, a C1-C14 monovalent heteroaralkyl radical, C(s)R’ or , in which R’ is a C1-C6 monovalent aliphatic radical, a C1-C6 monovalent heteroaliphatic radical, a monovalent aryl radical, a monovalent heteroaryl radical, a C1-C14 monovalent aralkyl radical, or a C1-C14 monovalent heteroaralkyl radical, provided that at least one of L1 and L2 is not a bond; each of W1, W2, W3, W4, W5, W6, W7, and W8, independently, is N or CR5, R5 being H, halo, cyano, amino, hydroxyl, nitro, sulfhydryl, a C1-C6 aliphatic radical, a C1-C6 heteroaliphatic l, or a iphatic radical; X is a bond, O, S, or NR6, R6 being H, a C1- C6 monovalent aliphatic l, a C1-C6 monovalent heteroaliphatic radical, a monovalent aryl radical, a monovalent heteroaryl radical, a C1-C14 lent aralkyl radical, or a C1-C14 monovalent heteroaralkyl radical; Y is a aryl ring or a heteroaryl ring; and Z is a therapeutic moiety.
In a particular aspect, the present invention provides a compound of formula (I) in which each of A1, A2, A3, A4, A5, and A6, independently, is a C1-C6 bivalent aliphatic radical; B1 is a C1-C6 bivalent aliphatic radical, a C1-C6 bivalent heteroaliphatic radical, a bivalent aryl l, or a bivalent heteroaryl radical; B2 is a bond, a C1-C6 nt aliphatic radical, a C1-C6 bivalent heteroaliphatic radical, a nt aryl radical, a bivalent heteroaryl radical, D1-NR 1-C(O)-D2, D1-C(O)NR 1-D2-NR 1’-C(O)-D3, D1-D2-C(O)-NR 1-C(O)-D3, or D1-D2-D3, each of D1, D2, D3, ndently, being a C1-C6 bivalent aliphatic radical, a C1-C6 bivalent heteroaliphatic radical, a bivalent aryl radical, a bivalent heteroaryl radical, a C1-C10 bivalent aralkyl radical, or a C1-C10 bivalent heteroaralkyl radical, and each of R1 and R1’, independently, being H, a C1-C6 bivalent heteroaliphatic radical, a bivalent aryl radical, a bivalent heteroaryl radical, or a C1-C10 bivalent aralkyl radical; L1 is a bond, NR2, NR2C(O), NR 2C(S), NR2CR 3R4, NR2SO 2, NR2C(O)NR 3, or NR2C(S)NR 3, each of R2, R3, and R4, independently, being H, a C1-C6 monovalent aliphatic radical, a C1-C6 monovalent aliphatic radical, a monovalent aryl l, a monovalent heteroaryl radical, a C1-C14 monovalent aralkyl radical, a C1-C14 monovalent aralkyl radical, C(s)R’, or C(O)R’, in which R’ is a C1-C6 lent aliphatic radical, a C1-C6 monovalent heteroaliphatic radical, a lent aryl radical, a monovalent heteroaryl radical, a C1-C14 monovalent l radical, or a C1-C14 monovalent heteroaralkyl radical; L2 is NR2, NR2C(O), NR2C(S), NR 2CR 3R4, NR2SO 2, )NR 3, NR2C(S)NR 3, or C(O), each of R2, R3, and R4, independently, being H, a C1-C6 monovalent aliphatic radical, a C1-C6 lent heteroaliphatic radical, a monovalent aryl l, a monovalent heteroaryl radical, a C1-C14 monovalent aralkyl radical, a C1-C14 monovalent heteroaralkyl radical, C(s)R’, or C(O)R’, in which R’ is a C1-C6 monovalent tic radical, a C1-C6 monovalent heteroaliphatic radical, a monovalent aryl radical, a monovalent heteroaryl radical, a C1-C14 monovalent aralkyl radical, or a C1-C14 monovalent heteroaralkyl radical; each of W1, W2, W3, W4, W5, W6, W7, and W8, independently, is N or CH; X is a bond, O, S, or NR6, R6 being H, a C1-C6 monovalent aliphatic radical, a C1-C6 monovalent heteroaliphatic radical, a monovalent aryl l, a monovalent heteroaryl radical, a C1-C14 monovalent aralkyl radical, or a C1-C14 monovalent heteroaralkyl radical; Y is an aryl ring or a heteroaryl ring; and Z is an anticancer (followed by page 3a) therapeutic moiety connected to L2 via a cleavable bond; wherein each of the aliphatic radical, the heteroaliphatic radical, the aralkyl radical, and the heteroaralkyl radical is unsubstituted or substituted with halo, cyano, amino, hydroxyl, nitro, sulfhydryl, C1-C6 alkoxy, C1-C6 alkylamino, C1-C12 dialkylamino, and C1-C6 haloalkyl; and each of the aryl radical and the heteroaryl radical is tituted or substituted with halo, cyano, amino, yl, nitro, sulfhydryl, a C1-C6 tic radical, a C1-C6 heteroaliphatic radical, or a haloaliphatic radical.
Each of the aliphatic radical, the heteroaliphatic radical, the aralkyl l, and the heteroaralkyl radical is unsubstituted or substituted with halo, cyano, amino, hydroxyl, nitro, sulfhydryl, C1-C6 alkoxy, C1-C6 alkylamino, C1-C12 dialkylamino, and C1-C6 haloalkyl; and each of the aryl radical and the aryl radical is unsubstituted or tuted with halo, cyano, amino, hydroxyl, nitro, sulfhydryl, a C1-C6 tic radical, a C1-C6 heteroaliphatic radical, or a haloaliphatic radical.
A subset of the compounds described above are those of formula (I), in which Y is Referring to formula (I) again, another subset are those, in which each of W1, W2, W3, and W4 is N, and each of W5, W6, W7, and W8 is CH; or in which each of W1, W2, W3, and W8 is N, and each of W4, W5, W6, and W7 is CH.
Still another subset are those of formula (I), in which each of A1, A2, A3, A5, A5, and A6 is methylene.
Further, in the compounds of formula (I), B1 can be ethylene, propylene, butylene, or ne; X can be O or NH; L2 can be C(O).
Also, in the above-described compounds, L1 can be a bond, NH, NHCH2, NHC(O), NHSO2, NHC(O)NH, [FOLLOWED BY PAGE 4] ONE?N\ W0 W0 0Y0 ){L v.32. ’ ){LNV’ , 3;va 052/ \/ ’ / ,N z ,N ’, / ‘7; /,N\}{, )2le , ,3 \/ IO CI / \ 0 S O/N\ O O o :QL/NVL‘Q ‘ / O EN “f N=N\ O O WN H 55; a0N / ’1‘;O 9% a” ‘5‘ ”E ”AGE O ’ ,or o a; ~91@NNOwONNH O In compounds of formula (I), Z, a therapeutic moiety, is formed from a eutic drug. It is ted to L2 Via a bond, e.g., an amide bond or an ester or thioester bond.
Upon release from formula (1) Via enzymatic hydrolysis, Z is converted to a therapeutic drug that exerts a cytotoxic effect, e.g., anti-proliferation.
Preferably, Z is an anticancer therapeutic moiety, an rus therapeutic moiety, an antibiotic therapeutic moiety, an immuno-stimulatory therapeutic moiety, an immuno- ssive therapeutic moiety, a therapeutic moiety for treating a cardiovascular disease, or a cytotoxic moiety. Examples of Z include, but are not limited to, The term “aliphatic” herein refers to a saturated or unsaturated, linear or branched, acyclic, cyclic, or polycyclic hydrocarbon moiety. Examples include, but are not limited to, alkyl, alkylene, l, alkenylene, alkynyl, alkynylene, cycloalkyl, cycloalkylene, cycloalkenyl, cycloalkenylene, cycloalkynyl, and cycloalkynylene es. The term “heteroaliphatic” herein refers to an aliphatic moiety containing at least one heteroatom selected from N, O, P, B, S, Si, Sb, Al, Sn, As, Se, and Ge. The term “haloaliphatic” herein refers to an aliphatic moiety substituted with one or more n atoms. The term “alkyl” WO 79299 herein refers to a straight or branched hydrocarbon group, containing 1—20 (e. g., 1—10 and l— 6) carbon atoms. Examples include methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, and t- butyl. The term “alkylene” refers to bivalent alkyl. Examples include —CH2—, —CH2CH2—, —CH2CH2CH2—, —CH2(CH3)CH2—, and —CH2CH2CH2CH2—. The term lkyl” refers to alkyl substituted with one or more halogen (chloro, , bromo, or idodo) atoms.
Examples include trifluoromethyl, bromomethyl, and 4,4,4-trifluorobutyl. The term “haloalkylene” refers to bivalent haloalkyl. The term “heteroalkylene” refers to a bivalent alkyl group, in which one or more carbon atoms are replaced by a heteroatom (e. g., 0, N, P, and S). The term “alkoxy” refers to an —O—alkyl group. Examples include methoxy, ethoxy, propoxy, and isopropoxy. The term “haloalkoxy” refers to alkoxy substituted with one or more halogen atoms.
The term “alkenyl” refers to a ht or branched hydrocarbon group, containing 2— (e. g., 2—10 and 2—6) carbon atoms and one or more double bonds.
The term “cycloalkyl” refers to a ted and lly unsaturated monocyclic, bicyclic, tricyclic, or tetracyclic hydrocarbon group having 3 to 12 carbons. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl. The term alkylene” refers to bivalent cycloalkyl.
The term “heterocycloalkyl” refers to a nonaromatic 5—8 membered monocyclic, 8—12 membered bicyclic, or 11—14 membered tricyclic ring system having one or more atoms (e. g., 0, N, P, and S). Examples of heterocycloalkyl groups include, but are not limited to, piperazinyl, imidazolidinyl, yl, pyrrolidinyl, dihydrothiadiazolyl, dioxanyl, morpholinyl, tetrahydropuranyl, and tetrahydrofuranyl. The term “heterocycloalkylene” refers to bivalent heterocycloalkyl.
The term “aryl” refers to a 6-carbon monocyclic, lO-carbon bicyclic, l4-carbon tricyclic aromatic ring system wherein each ring may have 1 to 5 substituents. Examples of aryl groups include phenyl, naphthyl, and anthracenyl. The term “arylene” refers to bivalent aryl. The term “aralkyl” refers to alkyl substituted with an aryl group. The term enyl” refers to alkenyl tuted with an aryl group.
The term oaryl” refers to an aromatic 5—8 membered monocyclic, 8—12 membered bicyclic, or 11—14 membered tricyclic ring system having one or more heteroatoms (e. g., 0, N, P, and S). Examples include triazolyl, oxazolyl, thiadiazolyl, tetrazolyl, pyrazolyl, pyridyl, furyl, olyl, benzimidazolyl, dinyl, thienyl, quinolinyl, indolyl, thiazolyl, and hiazolyl. The term “heteroaralkyl” refers to an alkyl group substituted with a heteroaryl group. The term “heteroaralkenyl” refers to an alkenyl group substituted with a aryl group. The term “heteroarylene” refers to bivalent heteroaryl.
The term “halo” refers to a fluoro, chloro, bromo, or iodo radical. The term “amino” refers to a radical derived from amine, which is tituted or mono-/di-substituted with alkyl, aryl, cycloalkyl, heterocycloalkyl, or heteroaryl. The term “alkylamino” refers to alkyl—NH—. The term “dialkylamino” refers to alkyl—N(alkyl)—.
The term “acyl” refers to C(O) alkyl, C(O) aryl, C(O) cycloalkyl, —C(O)— heterocycloalkyl, or heteroaryl.
Alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, alkoxy, and aryloxy mentioned herein include both substituted and unsubstituted moieties. Examples of substituents include, but are not limited to, halo, hydroxyl, amino, cyano, nitro, mercapto, alkoxycarbonyl, amido, carboxy, alkanesulfonyl, alkylcarbonyl, carbamido, carbamyl, carboxyl, thioureido, thiocyanato, sulfonamido, alkyl, alkenyl, alkynyl, alkyloxy, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl, in which alkyl, l, alkynyl, alkyloxy, aryl, heteroaryl cycloalkyl, and heterocycloalkyl may further substituted. , the term “compound” refers to the compounds bed above, as well as their salts, solvates, and metal complexes. A salt can be formed between an anion and a positively d group (e. g., amino) on a compound; examples of a suitable anion include chloride, bromide, iodide, sulfate, e, ate, citrate, methanesulfonate, trifluoroacetate, acetate, malate, tosylate, tartrate, fumurate, glutamate, glucuronate, lactate, ate, and maleate. A salt can also be formed between a cation and a negatively charged group; examples of a suitable cation include sodium ion, potassium ion, magnesium ion, calcium ion, and an ammonium cation such as tetramethylammonium ion. A salt further includes those containing quaternary nitrogen atoms. A solvate refers to a complex formed n an active compound and a pharmaceutically acceptable solvent. Examples of a pharmaceutically acceptable solvent include water, ethanol, panol, ethyl acetate, acetic acid, and ethanolamine. A metal complex can be formed of a compound and a metal ion.
The metal ion is a cation having two or more charges. The metal complex is typically formed via ion of a metal ion and a compound of formula (1). Examples of the metal ion include Zn2+, Cu2+, Ca2+, Mg2+, Mn2+, Ni2+, Coz+, Fe2+, Cd2+, and a combination thereof.
This invention also es use of one of the above-described compounds for the manufacture of a medicament for treating PS-related conditions. Thus, this invention also relates to use of such a compound for treating a PS-related condition by administering to a subject in need of the treatment an effective amount of a compound of this invention and an effective amount of one or more other active agents.
Also within the scope of the present invention is a pharmaceutical composition containing a pharmaceutically acceptable carrier and a complex of a metal ion and one of the compounds of formula (I) described above.
The pharmaceutical composition can further contain another eutic agent for treating PS-related conditions. Active agents include, but are not d to, immunomodulatory agents, such as interferons 0t, B, and y; antiviral agents such as ribavirin and dine; therapeutic agents target in any PS-related conditions. Such an active agent and a compound of this invention may be d to a subject at two separate times or simultaneously but in two dosage forms. Alternatively, they can be combined in a composition as described above for use as a single dosage form.
A composition for oral administration can be any orally acceptable dosage form including capsules, tablets, emulsions and aqueous sions, dispersions, and solutions.
In the case of tablets, commonly used carriers include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions or emulsions are administered orally, the active ingredient can be suspended or dissolved in an oily phase combined with emulsifying or ding . If desired, certain sweetening, flavoring, or coloring agents can be added. Oral solid dosage forms can be prepared by spray dried techniques; hot melt extrusion strategy, micronization, and nano milling technologies.
A nasal aerosol or inhalation composition can be ed according to techniques well known in the art of pharmaceutical formulation. For example, such a composition can be prepared as a solution in saline, employing benzyl l or other le preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing agents known in the art. A composition having an active compound can also be administered in the form of suppositories for rectal administration.
The carrier in the ceutical composition must be “acceptable” in the sense that it is compatible with the active ient of the ition (and preferably, capable of stabilizing the active ingredient) and not deleterious to the t to be treated. One or more solubilizing agents can be utilized as pharmaceutical excipients for delivery of an active compound. Examples of other carriers include colloidal silicon oxide, magnesium stearate, cellulose, sodium lauryl sulfate, and D&C Yellow #10.
Still within the scope of this invention is a method of treating a condition associated with cells containing inside-out phosphatidylserine. In this condition, normally intracellular phosphatidylserine is d on the outer e of the cells. The method includes stering an effective amount of one of the compounds described above to a subject in need thereof. The compound of formula (I) can be administered as a complex formed of the compound and a metal ion having two or more positive charges (e.g., Zn2+). The condition exists in viral infection, bacterial infection, inflammatory e, cancer, misregulation of cell death in organ transplant, ulation of cell death in neurodegenerative disease, and misregulation of cell death in cardiovascular disease.
The term “treating” refers to application or administration of the compound to a subject with the e to cure, alleviate, relieve, alter, remedy, improve, or affect the disease, the symptom, or the predisposition. “An effective amount” refers to the amount of the compound which is required to confer the desired effect on the subject. ive amounts vary, as recognized by those skilled in the art, depending on route of administration, excipient usage, and the possibility of co-usage with other therapeutic treatments such as use of other active agents.
The details of one or more ments of the invention are set forth in the description below. Other features, objects, and advantages of the invention will be apparent from the description and from the claims.
DETAILED DESCRIPTION Shown below are 51 exemplary compounds synthesized following the ures described in Examples 1—51 and tested following the procedures bed thereafter.
All compounds listed below include their racemates (i.e., equal amounts of left- and handed enantiomers of a chiral molecule) unless otherwise specified.
Compound Compound ures numbers WO 79299 WO 79299 WO 79299 WO 79299 WO 79299 WO 79299 WO 79299 WO 79299 WO 79299 WO 79299 WO 79299 N N /N/N/N/N \l\|\|\| N N PNPNPNPN / / / / WO 79299 WO 79299 The nds of this invention can be prepared by synthetic methods well known in the art. See R. Larock, Comprehensive Organic Transformations (2ndl Ed., VCH Publishers 1999); P. G. M. Wuts and T. W. Greene, Greene’s Protective Groups in Organic Synthesis (4th Ed., John Wiley and Sons 2007); L. Fieser and M. Fieser, Fieser and Fieser’s Reagents for Organic Synthesis (John Wiley and Sons 1994); and L. te, ed., Encyclopedia of ts for Organic Synthesis (2ndl ed., John Wiley and Sons 2009) and uent editions thereof.
The syntheses of Compounds l-51 and their analytical data are shown below.
S nthesis of Di icol lamine Intermediate DPA structure shown below O/\/\/NH2 N N /N/N/N/N | I l I \ \ \ \ A dipicolylamine intermediate DPA was ed according to the following steps: OH OH TsOH + ”x“ —> HO OH MeOH 60°C /0 O\ O O 2,2-dimethoxypropane O O -hydroxyisophthalic acid dimethyl 5-hydroxyisophthalate To a stirred solution of 5-hydroxyisophthalic acid (25 g, 137 mmol, 1 eq.) in 200 mL of methanol at room temperature, 2,2-dimethoxypropane (1.2 eq.) and p-toluenesulfonic acid (TsOH, 0.2 eq.) were added. After stirring at 60 0C, methanol was removed by evaporation.
The resultant crude mixture was partitioned in ethyl e (EtOAc)/water. The organic layer was dried with magnesium sulfate (MgSO4) and concentrated to give a crude product, which was purified by -column chromatography to yield 2,2-dimethoxypropane ( 23 g, 79% ). 1H NMR (300 MHz, CDC13) 8 3.96 (s, 6H), 7.77 (s, 2H), 8.26 (s, 1H). 0“ OH /O 0\ dry THF 0 O OH OH dimethyl 5- (5-hydroxy—1,3-phenylene)dimethanol hydroxyisophthalate To a solution of dimethyl 5-hydroxyisophthalate (28 g, 133 mmol, 1 eq.) in 1400 mL of dry tetrahydrofuran (THF) at ice-bath temperature, lithium aluminum hydride (LiAlH4 or LAH, 4 eq.) was slowly added with stirring. The resultant mixture was allowed warm to 40 °C, stirred at this temperature for 16 hours, and then was added an ammonium chloride (NH4Cl) aqueous solution to quench the on. After stirring for 1.5 hours, the e was filtered with celite, washed with THF. Organic volatiles were evaporated and the residue was partitioned between water and EtOAc. The aqueous phase was extracted with EtOAc three times. The combined EtOAc layers were dried over MgSO4 to give product (5- hydroxy-l,3-phenylene)dimethanol (17 g, 82%). 1H NMR (400 MHz, CDC13) 8 4.57 (s, 6H), 6.71 (s, 2H), 6.80 (s, 1H).
Step 3 K2C03 N N — ’ O Br/VV CHgCN OH OH _ _ _O 2-(4—bromobutyl)Isomdollne-1,3-dlone_ (5-hydroxy-1,3- OH OH pheny'ene)d'metha“°' 2-(4-(3,5-bis(hydroxymethy|) phenoxy)butyl)isoindoline -1,3-dione To a stirred solution of roxy-1,3-phenylene)dimethanol (7.27 g, 47 mmol, 1 eq.) in 200 mL of acetonitrile at room temperature, 2-(4-bromobutyl)isoindoline-1,3-dione (1.2 eq.) and potassium carbonate (K2CO3, 2 eq.) were slowly added. The resultant mixture was then heated at reflux for 8 hours, after which the volatiles were removed, and the e was partitioned between EtOAc and water. The EtOAc layer was washed with brine and dried over MgSO4. ation of EtOAc gave 2-(4-(3,5- bis(hydroxymethyl)phenoxy)butyl)isoindoline-1,3-dione (13 g, 78% yield). 1H NMR (400 MHz, CDC13) a 1.67-1.65 (m, 2H), 1.86-1.81 (m, 2H), 3.75 (t, J = 5.6 Hz, 2H), 4.00 (t, J = 6.0 Hz, 2H), 4.63 (s, 2H), 4.64 (s, 2H), 6.81 (s, 2H), 6.91 (s, 1H), 7.71— 7.69 (m, 2H), 7.84-7.82 (m, 2H).
Step 4 O O O/\/\/N CBr4 pph3 N o —> o CH2C|2 r.t.
OH OH Br Br 2-(4-(3,5-bis(bromomethyl)phenoxy) 2-(4-(3,5-biS(hydr0Xymethyl) butyl)isoindoline-1,3-dione y)butyl)isoindo|ine-1,3-dione To a stirred solution of 2-(4-(3,5-bis(hydroxymethyl)phenoxy)butyl)isoindoline-1,3- dione (7.44 g, 21 mmol, 1 eq.) in 420 mL of anhydrous dichloromethane (CHzClz) in an ice bath, triphenylphosphine (PPh3, 2.3 eq.) and carbon tetrabromide (CBr4, 4.5 eq.) were slowly added. The resultant reaction e was allowed to warm to room temperature, stirred for 16, and added methanol (MeOH) to quench the reaction. After CHzClz and MeOH were evaporated, the residue was partitioned in /water. The CHzClz layer was dried over MgSO4 and concentrated by evaporation to give a crude product, which was purified by silica-column chromatography to yield product (5.2 g, 51%). 1H NMR (300 MHz, CDC13) 8 1.90-1.81 (m, 4H), 3.77 (t, J = 6.6 Hz, 2H), 4.00 (t, J = .7 Hz, 2H), 4.41 (s, 4H), 6.82 (s, 2H), 6.98 (s, 1H), 7.74-7.70 (m, 2H), 7.86-7.84 (m, 2H).
Step 5 O O/\/\/ H O O/\/\/N K2C03 + —> I I N N \ \ Br Br bis(pyridinylmethy|)amine / N / N N / N 2-(4-(3,5- I I I , bis(bromomethyl)phenoxy) \ \ \ buty|)isoindoline-1,3-dione To a stirred solution of 2-(4-(3,5-bis(bromomethyl)phenoxy)butyl)isoindoline-1,3- dione (3.86 g, 8.06 mmol, 1 eq.) in 25 mL of dry dimethylformamide (DMF) at room temperature, bis(pyridinylmethyl)amine (2 eq.) and K2CO3 (5 eq.) were slowly added.
After stirring for 16 hours, DMF was evaporated. The resultant crude mixture was partitioned in CHzClz/water. The CHzClz layer was dried over MgSO4, and trated to give a crude product, which was purified by silica-column tography to yield product A (4.8 g, 83%). 1H NMR (400 MHz, CDC13) 8 1.89-1.82 (m, 4H), 3.61 (s, 4H), 3.78-3.74 (m, 10H), 3.96 (t, J = 6.0 Hz, 2H), 6.82 (s, 2H), 7.03 (s, 1H), .08 (m, 4H), 7.63-7.53 (m, 8H), 7.69-7.67 (m, 2H), 7.82-7.80 (m, 2H), 8.48 (d, J = 4.8 Hz, 4H).
Step 6 O/WNH2 O/VVN HzN-NH2 + H'.q EtOH 100°C N N //N/ I I IN I I” \ \\ \ To a stirred solution of compound A (5.9 g, 8.23 mmol, 1 eq.) in 200 mL of ethanol (EtOH) at room temperature, hydrazine (HzN-NHZ, 10 eq.) was slowly added. The resultant reaction mixture was stirred for 16 hours, heated at reflux for 2 hours, and then cooled to room temperature. Removal of EtOH gave a crude mixture, which was extracted by CHzClz twice. The CHzClz solutions were ed, dried over MgSO4, and concentrated to afford DPA (4.1 g, 85%). 1H NMR (300 MHz, CDC13) 8 1.65-1.55 (m, 2H), 1.84-1.75 (m, 2H), 2.75 (t, J = 6.9 Hz, 2H), 3.56 (s, 4H), 3.78 (s, 8H), 3.94 (t, J = 6.6 Hz, 2H), 6.84 (s, 2H), 7.04 (s, 1H), 7.13- 7.08 (m, 4H), 7.63-7.55 (m, 8H), 8.48 (d, J = 4.5 Hz, 4H).
S nthesis of nine DPA s DL-l to DL-9 DL-l aaaa DPA (400 mg, 0.681mmol, 1 eq.) and triethylamine (1 mL) were dissolved in CHzClz (40 mL), followed by addition of ethyl 4-chlorooxobutanoate at 0°C. The resultant solution, after stirring at 0°C for 2 hours, was washed with a saturated ammonium chloride aqueous solution three times (3 X 40 ml). The CHzClz layers were dried over MgSO4 and concentrated under reduced pressure to yield DL-l. 1H NMR (400 MHz, CDC13) 8 .19 (m, 3H), 1.70-1.67 (m, 2H), 1.84-1.78 (m, 2H), 2.44 (t, J = 6.4 Hz, 2H), 2.77 (t, J = 6.4 Hz, 2H), 3.43-3.29 (m, 2H), 3.64 (s, 4H), 3.79 (s, 8H), 3.96 (t, J = 6 Hz, 2H), 4.16-4.09 (m, 2H), 6.83 (s, 2H), 7.05 (s, 1H), 7.14-7.11 (m, 4H), 7.64-7.57 (m, 8H), 8.50 (d, J = 4.4 Hz, 4H).
DL-2 aaaa DL-l (487 mg, 0.68 mmol, 1 eq.) was dissolved in MeOH (4 mL) and a LiOH aqueous solution (4 mL, 0.5 N), and then stirred at room temperature for 15 hours. After removal of MeOH, the resultant residue was extracted with CHzClz (100 mL). The CHzClz solution was then washed with a saturated ammonium chloride aqueous solution twice (2 X 100 mL), dried over MgSO4, and trated under reduced pressure to yield DL-2 (380 mg). 1H NMR (300 MHz, CDC13) 8 .67 (m, 2H), 1.82-1.79 (m, 2H), 2.48 (m, 2H), 2.68 (m, 2H), 3.34 (m, 2H), 3.63 (s, 4H), 3.80 (s, 8H), 3.96 (t, J = 8 Hz, 2H), 6.83 (s, 2H), 6.90 (s, 1H), 7.15-7.11 (m, 4H), 7.66-7.54 (m, 8H), 8.51 (d, J = 4.8 Hz, 4H).
DL-3 N N To a solution of DPA (400 mg, 0.68 mmol, 1 eq.) in CHzClz (40 mL) were added triethylamine (2 mL) and methyl 3-(chlorocarbonyl) benzoate at 0°C. The resultant reaction mixture, after stirring at 0°C for 2 hours, was diluted with CHzClz (100 mL). The CHzClz on was washed with a saturated s solution of NH4Cl twice (2 X 100 mL), dried over MgSO4, and concentrated under reduced pressure to give a crude product, which is purified by column chromatography (silica gel; MeOH : CHzClz = 1 : 13) to yield DL-3 (280 mg, 55%). 1H NMR (400 MHz, CDC13) 8 1.91-1.84 (m, 4H), 3.58-3.54 (m, 2H), 3.63 (s, 4H), 3.78 (s, 8H), 3.91 (s, 3H), 4.02 (t, J = 5.2 Hz, 2H), 6.85 (s, 2H), 7.05 (s, 1H), 7.13-7.10 (m, 4H), 7.50-7.46 (m, 1H), 7.63-7.56 (m, 8H), 8.00 (d, J = 7.6 Hz, 1H), 8.12 (d, J = 8 Hz, 1H), 8.36 (s, 1H), 8.49 (d, J = 4.4 Hz, 4H). 2015/031386 DL-4 N N |\N|\N ‘N ‘N DPA (1 g, 1.7 mmol, 1 eq.) and methyl 4-formylbenzoate (840 mg, 5.12 mmol, 3 eq) were dissolved in MeOH (20 mL) and stirred at 65°C for 15 hours. After the solution was cooled to 0°C, sodium borohydride (1 g, 26 mmol, 15 eq) was added. The mixture was stirred at 0 0C for another hour. Removal of MeOH under d pressure gave a residue, which was extracted with CHzClz (100 mL). The CHzClz solution was washed with a saturated s solution of NH4Cl twice (2 X 100 mL), dried over MgSO4, and concentrated under reduced pressure. The resultant residue was purified by column chromatography (silica gel; MeOH : CHzClz = 1 : 9) to yield DL-4 (700 mg, 56%). 1H NMR (300 MHz, CDC13) 8 1.90-1.68 (m, 4H), 2.71 (t, J = 7.2 Hz, 2H), 3.63 (s, 4H), 3.79 (s, 8H), 3.82 (s, 2H), 3.86 (s, 3H), 3.95 (t, J = 6 Hz, 2H), 6.83 (s, 2H), 7.04 (s, 1H), 7.14-7.09 (m, 4H), 7.40 (d, J = 8.1 Hz, 2H), 7.63-7.55 (m, 8H), 7.98 (d, J = 8.1 Hz, 2H), 8.48 (d, J = 4.2 Hz, 4H).
DL-S N N |\N|\N ‘N ‘N DL-4 (600 mg, 0.82 mmol, 1 eq) and di-tert-butyl dicarbonate (360 mg, 1.65 mmol, 2 eq) were dissolved in CHzClz (60 mL) and stirred at room temperature for 15 hours. After CHzClz was removed, a residue was obtained and purified by column chromatography (silica gel; MeOH : CHzClz = 1 : 13) to yield DL-5 (550 mg, 81%). 1H NMR (300 MHz, CDC13) 8 1.45-1.40 (m, 9H), 1.72 (m, 2H), 1.89 (m, 2H), 3.31- 3.21 (m, 2H), 3.64 (s, 4H), 3.79 (s, 8H), 3.89 (s, 3H), 3.92 (m, 2H), 4.47 (m, 2H), 6.82 (s, 2H), 7.07 (s, 1H), .09 (m, 4H), 7.27 (d, J = 9 Hz, 2H), 7.63-7.56 (m, 8H), 7.98 (d, J = 8.7 Hz, 2H), 8.50 (d, J = 4.8 Hz, 4H).
DL-6 H O N N |\N|\N ‘N \N DL-4 (300 mg, 0.41 mmol) was dissolved in MeOH (3 mL) and an aqueous LiOH solution (3 mL, 0.5 N). The resultant mixture was stirred at room temperature for 15 hours.
Removal of MeOH gave a residue, which was ted with CHzClz (100 mL). The CHzClz solution was then washed with a saturated aqueous solution of NH4Cl twice (2 x 100 mL), dried over MgSO4, and concentrated under reduced re to yield DL-6 (260 mg, 88%).
DL-7 N N |\N|\N \N \N DL-5 (550 mg, 0.66 mmol) was dissolved in MeOH (6 mL) and an aqueous LiOH solution (6 mL, 0.5 N). The resultant mixture was stirred at room temperature 15 hours.
MeOH was removed to give a residue, which was extracted with CHzClz (100 mL). The CHzClz solution was washed with a saturated aqueous solution of NH4Cl twice (2 x 100 ml), dried over MgSO4, and concentrated under reduced pressure to yield DL-7 (480 mg, 89%). 1H NMR (300 MHz, CDC13) 8 1.50-1.26 (m, 13H), 3.29-3.23 (m, 2H), 3.65 (s, 4H), 3.81 (m, 10H), 4.53 (s, 2H), 6.77 (s, 2H), 6.93 (s, 1H), 7.15-7.11 (m, 4H), 7.38 (m, 2H), 7.63- 7.53 (m, 8H), 8.10 (d, J = 7.8 Hz, 2H), 8.54 (d, J = 4.2 Hz, 2H) DL-8 N N \N \N \N \N I I DL-6 (260 mg, 0.36 mmol, 1 eq), K2CO3 (745 mg, 5.40 mmol, 15 eq), 4-chloro nitrobenzo[c][1,2,5] oxadiazole (100 mg, 0.50 mmol), and CHzClz (30 mL) were mixed and stirred at 40°C for 15 hours. The resultant reaction mixture was then extracted with CHzClz (100 mL). uently, the CHzClz solution was washed with water twice (2 x 100 mL), dried over MgSO4, and concentrated under reduced re. The resultant residue was purified by column chromatography (silica gel; MeOH : CHzClz = 1 : 1) to yield DL-8 (200 mg, 63%) 1H NMR (400 MHz, DMSO) 8 1.36 (m, 1H), 1.69-1.51 (m, 3H), 3.13 (m, 2H), 3.54 (s, 4H), 3.66 (m, 10H), 3.98 (m, 2H), 6.68 (m, 1H), 6.79 (d, J = 4.4 Hz, 2H), 7.04 (d, J = 5.6 Hz, 1H), .17 (m, 4H), 7.35-7.25 (m, 2H), 7.54-7.51 (m, 4H), 7.71-7.63 (m, 4H), 7.86 (d, J = 8.4 Hz, 2H), 8.38 (m, 1H), 8.44 (d, J = 5.2 Hz, 4H).
DL-9 N N I‘NPN ‘N ‘N A solution of DL-3 (0.37 mmol) in MeOH (3 mL) and an aqueous LiOH on (3 mL, 0.5 N) was d at room temperature for 15 hours. MeOH was removed under reduced pressure to give a residue, which was extracted with CHzClz (100 mL). The CHzClz solution was then washed with a saturated aqueous solution of NH4Cl twice (2 x 100 mL), dried over MgSO4, and concentrated under reduced pressure to yield DL-9 (240 mg, 88%). 1H NMR (300 MHz, CDC13) 8 1.97-1.86 (m, 4H), 3.64 (m, 6H), 3.80 (s, 8H), 4.14 (m, 2H), 7.08 (s, 2H), 7.20-7.12 (m, 5H), .51 (m, 8H), 8.21 (d, J = 7.2 Hz, 2H), 8.47 (s, 1H), 8.56 (d, J = 4.8 Hz, 4H).
EXAMPLE 1: ation of Compound 1 Compound 1 of this invention was prepared following the procedure bed below.
To a solution of DL-2 (200 mg, 0.29 mmol, 1 eq) in DMF (20 mL) were added 4,11- diethyl-4,9-dihydroxy-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14 (4H,12H)-dione (170 mg, 0.44 mmol, 1.5 eq), hydroxybenzotriazole (117 mg, 0.87 mmol, 3 eq), N-(3- dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (170 mg, 0.87 mmol, 3 eq), and N-methylmorpholine (175 mg, 1.74 mmol, 6 eq). After stirring at room temperature for 15 hours, the resultant reaction mixture was extracted with CHzClz (300 mL). Subsequently, the CHzClz solution was washed with a saturated aqueous solution of NaHC03 (300 mL) and water (5 x 300 mL), dried over MgSO4, and then concentrated under reduced re to give a residue. Compound 1 (130 mg, 42 %) was obtained by purifying the residue with column chromatography (silica gel; MeOH : CHzClz = 1 : 13). 1H NMR (400 MHz, CDC13) 8 1.01 (t, J :72 Hz, 3H), 1.34 (t, J = 7.6 Hz, 3H), 1.74- 1.67 (m, 2H), 1.94-1.77 (m, 4H), 2.62 (t, J = 6.4 Hz, 2H), 3.00 (t, J = 6.4 Hz, 2H), 3.11-3.06 (m, 2H), 3.39-3.34 (m, 2H), 3.61 (s, 4H), 3.76 (s, 8H), 3.92 (t, J =j = 6.0 Hz, 2H), 5.20 (s, 2H), 5.28 (d, J = 16.4 Hz, 1H), 5.71 (d, J = 16.4 Hz, 1H), 6.77 (s, 2H), 7.04 (s, 1H), 7.12-7.09 (m, 4H), 7.63-7.50 (m, 10H), 7.77 (d, J = 2.4 Hz, 1H), 8.14 (d, J = 9.2 Hz, 1H), 8.47 (d, J = 4.8 Hz, 4H). Mass: (EM+2H+)/2: found 532.27.
EXAMPLES 2-51: Preparation of Compounds 2-51 The synthesis of Compound 42 is described immediately below. As for compounds 2-41 and 43-51, they were obtained following a similar procedure or a ure similar to that described in Example 1 above.
Scheme 1 below depicts a synthetic sequence of ing a linker, i.e., 42-Linker, from commercially available B via intermediates C and D.
S nthesis of 42-Linker alinker for use in re arin Com ound 42: 0 o / 01—O a H L, N —’ /—/ /—/ O/—/ ofo of ofo’ /_/ /_/ /_/ 800‘” 800‘” H2N B C D ker Scheme 1. Synthesis of a : (a) I. B0020, DCM, 3 hours; and II. Methyl 4- formylbenzoate, NaBH4, MeOH, room temperature, 1 hour; (b) 1-(4- Chlorophenyl)cyclo-hexanecarbonyl chloride, DCM, room temperature, 1 hour; (0) 4 M HCl in 1,4-dioxane, MeOH, room temperature, 12 hours.
Synthesis ofcompound C: To a solution of the commercial available B (15 g, 101.21 mmol) in CHzClz (300 mL) was added trimethylamine (TEA, 1 eq.), followed by di-tert-butyl dicarbonate (BoczO, 0.5 eq). The reaction was stirred at room temperature for 3 hours, after which time the solvent was removed in vacuo. The crude was partitioned between H20 and DCM. The layers were separated, and the cs were dried over sodium sulfate (NazSO4) and concentrated to a transparent oil. To the transparent oil in 200 ml of MeOH was added methyl 4-f0rmylbenz0ate (15 g, 91.35 mmol, 0.9 eq.). The sodium borohydride (3.7 g, 97.80 mmol, 0.9 eq.) was added after stirring at room temperature for 3 hours. The MeOH was removed and the residue dissolved in 200 ml CHzClz. The protonated product was extracted from CHzClz with 200 ml NH4Cl(aq). The organic layers were ed, dried with , ed and the solvent evaporated. Purification of the crude residue by flash chromatography on silica gel eluting with EtOAc/Hexane(2: 1) to give 12.03 g of t C as a transparent oil (30.36 mmol, 30%).
Synthesis ofcompound D: To the compound C (12.03 g, 30.36 mmol) in 200 ml CHzClz was added hlorophenyl)cyclo- hexanecarbonyl chloride(11.57 g and , 45.54 mmol, 1.5 eq.) triethylamine(10 ml, 43.08 mmol). The on was stirred for 2 hours at room temperature.
The protonated product was extracted from 200 ml CHzClz with 200ml NH4Cl(aq). The organic layer was dried with NaZSO4, filtered, and the solvent evaporated. Purification of the crude residue by flash chromatography on silica gel eluting with EA/Hexane(2: 1) gave 13.11 g of compound D as a transparent oil (21.25 mmol, 70%).
Synthesis ofcompound 42-Linker: To the compound D (13.11 g, 21.25 mmol) in MeOH (200 mL) was added 4 M hydrochloric acid (HCl) in dioxane (10 mL). The reaction was stirred at room temperature for 2 hours, after which time it was trated in vacuo. The crude was partitioned n NaH4Cl(aq) and DCM. The layers were separated, and the organics were dried (NaZSO4) and concentrated to obtain compound 46-Linker as a transparent oil (7.68 g, 14.87 mmol, 70%).
Scheme 2 below depicts a synthetic sequence of preparing Compound 42 from the ediate DPA via intermediates E, F, G, H, and I.
S nthesis of Com ound 42: . 3309 C 009 NFC???0 CW) :29 09 09.9009 98 ofN o OH / 0 \ 0 \/HNf 7 /l Cwb C85 “0 Compound 42 Scheme 2. Synthesis of Compound 42. (a) Methyl 4-formylbenzoate, NaBH4, MeOH, room temperature, 1 hour (90%); (b) 1-(4-Chlorophenyl)cyclo-hexanecarbonyl de, CHzClz, room temperature, 1 hour (90%); (c) 0.5M LiOqu), MeOH, room temperature, 15 hours (90%); (d) 42-Linker, HOBt, EDCI, NMM, room temperature, hours (40%); (e) 0.5M LiOH(aq), MeOH, room temperature, 15 hours (82%); (f) HOBt, EDCI, NMM, 4,11-diethyl-4,9-dihydroxy-1H- pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14-(4H,12H)-dione, room temperature, 15 hours (30%).
Synthesis ofcompound E: To the above-mentioned DPA (10 g,17.01 mmol) in 200 ml of MeOH was added methyl 4-formylbenzoate (5 g, 30.45 mmol, 1.8 eq.). Sodium borohydride (3.7 g, 97.80 mmol, 5.7 eq.) was added after stirring at room temperature for 3 hours. MeOH was removed and the residue was dissolved in 200 ml of CHzClz. The protonated t was extracted from CHzClz with 200 ml of 1M HCl(aq). The s layer was neutralized and the product was extracted into 200 ml of CHzClz. The c layers were combined, dried with NaZSO4, and filtered, and the solvent was evaporated to give 11.26 g of product E as a yellow oil (15.30 mmol, 90%). 1H NMR (300 MHz, CDC13)C 8 1.70-1.78(m, 2H), 1.80- 1.86(m, 2H), 2.71(t, J = 6.8 Hz, 2H ), 3.62(s, 4H), 3.78(s, 8H), 3.87(s, 2H), , 3H), 3.95(t, J = 6.4 Hz, 2H), 6.82(s, 2H), 7.03(s, 1H), 7.03-7.13(m, 4H), 7.40(d, J = 8.0 Hz, 2H), 7.55-7.62(m, 8H), 7.98(d, J = 8.0 Hz, 2H), , J = 8.0 Hz, 2H).
Synthesis ofcompound F: To the nd E (11.26 g,15.30 mmol) in 200 ml of CHzClz was added l-(4-Chlorophenyl)cyclo- hexanecarbonyl chloride (7.71 g, 30.00 mmol, 2 eq.) and trimethylamine (5ml, 21.54 mmol). The reaction was stirred for 2 hours at room temperature. The protonated product was extracted from 200 ml of CHzClz with 200ml of 1M HCl(aq). The aqueous layer was neutralized and the product was ted into 200 ml of CHzClz. The organic layer was dried with NaZSO4, filtered, and the solvent was evaporated to give 13.17 g of product F as a yellow oil (13.77 mmol, 90%). 1H NMR (300 MHz, C 8 1.62(brs, 12H), 2.24(brs, 2H), 2.90(brs, 1H), 3.23(brs, 1H), 3.63(s, 4H), 3.78(s, 8H), 3.88(s, 3H), .3.92(m, 2H), 4.02-4.14(m, 2H), , 2H), 6.95-7.39(m, 11H), 7.55- 7.63(m, 8H), 7.93(d, J = 7.2 Hz, 2H ), 8.48(d, J = 6.4 Hz, 4H).
Synthesis ofcompound G: To the compound F (13.17g, 13.77 mmol) in 300 ml of MeOH was added 50 ml of 0.5M q). The reaction mixture was stirred at room temperature for hours. The solvent was removed and the residue was redissolved in CHzClz. The insoluble residue was filtered off. The filtrate was washed with water, dried over MgSO4(S) and the t was d under vacuum. The product G was obtained as yellowish powder (11.68 g, 12.39 mmol, 90% ), which was directly used for the next step.
Synthesis ofcompound H: A solution containing G (11.68 g, 12.39 mmol) in 40 ml of DMF was heated to 40 OC. EDCI ( 2 g, 12.8 mmol) and HOBt ( 2g, 14.8 mmol) were added and the resulting on was allowed to stir at room temperature for 30 minutes, compound 42- Linker, 4-({ {2-[2-(2-Amino-ethoxy)-ethoxy]-ethyl}-[1-(4-chloro-phenyl)- cyclohexanecarbonyl]-amino}-methyl)-benzoic acid methyl ester (9.6 g, 18.58 mmol) was added ed by addition of N-methylmorpholine (NMM, 5ml, 45.5 mmol). The reaction was stirred at room temperature for 15 hours, after which time it was diluted with H20. The aqueous solution was separated and extracted with 200 ml of CHzClz. The ed ts were washed with brine (4 x 100 mL), dried over NaZSO4(S), filtered, and ated.
Purification of the crude residue by flash chromatography on pH=7 silica gel eluting with MeOH/CH2C12(1:9) gave rise to ester compound H (7.14 g, 4.95 mmol, 40%).
Synthesis ofcompound I: To the compound H (7.14 g, 4.95 mmol) in 200 ml of MeOH was added 30 ml 0.5M LiOH(aq). The on mixture was stirred at room temperature for 15 hours. The solvent removed and the residue was redissolved in 100 ml CHzClz. The insoluble residue filtered off. The filtrate was washed with water, dried over MgSO4(S) and the solvent removed under vacuum. The product compound I was obtained as white powder (5.83 g, 4.08 mmol, 82%), which was directly used for the next step.
Synthesis of Compound 42: A solution of I (5.83 g, 4.08 mmol) in 20 ml of DMF was heated to 40 OC. 1-Ethyl(3-dimethylaminopropyl)carbodiimide (EDCI, 1 g, 6.4 mmol, 1.5 eq.) and hydroxybenzotriazole (HOBt, 1 g, 7.2 mmol, 1.7 eq.) were added and the reaction allowed to stir. After stirring at room temperature for 30 min, 4,11-diethyl-4,9-dihydroxy-1H- pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14-(4H,12H)-dione (3 g, 7.64 mmol, 1.87 eq.) was added followed by N—methylmorpholine (NMM, 5ml, 45.5mmol). The reaction was stirred at room temperature for 15 hours, after which time it was diluted with H20. The aqueous solution was separated and extracted with 100 ml of . The combined extracts were washed with brine (4 x 100 mL), dried over NaZSO4(S), filtered, and evaporated.
Purification of the crude residue by flash chromatography on pH=7 silica gel g with MeOH/CHzClz (0.5:9.5) gave rise to white power ester Compound 42 (2.21 g, 1.22 mmol, %).
Compound 2 was prepared from DL-9 at a yield of 40%. 1H NMR (400 MHz, CD3OD) 8 1.02 (t, J = 7.6 Hz, 3H), 1.39 (t, J = 7.6 Hz, 3H), 1.93-1.98 (m, 6H), 3.21-3.23 (m, 2H), 3.57 (m, 2H), 3.78 (s, 4H), 3.96 (d, J = 16 Hz, 4H), 4.11 (m, 2H), 4.34 (d, J = 16 Hz, 4H), 5.32 (s, 2H), 5.38 (d, J = 16.4 Hz, 1H), 5.58 (d, J =16.4 Hz, 1H), 6.72 (s, 1H), 6.81 (s, 2H), 7.60 (d, J = 7.6 Hz, 4H), 7.65 (s, 1H), 7.67-7.76 (m, 4H), 8.11-8.22 (m, 6H), 8.39 (d, J = 7.2 Hz, 1H), 8.69 (d, J = 4 Hz, 4H). Mass: (EM+2H+)/2: found 555.27.
Compound 3: 1H NMR (400 MHz, DMSO) 8 0.86 (t, J = 6.8 Hz, 3 H), 1.24 (t, J = 7.6 Hz, 3H), 1.45 (m, 4H), 1.71 (m, 4H), 1.85 (m, 2H), 2.67 (m, 2H), 3.14-3.16 (m, 2H), 3.55 (s, 4H), 3.67 (s, 8H), 3.94 (m, 2H), 5.30 (s, 2H), 5.42 (s, 2H), 6.80 (s, 2H), 7.04 (s, 1H), 7.20- 7.23 (m, 4H), 7.31 (s, 1H), 7.54 (d, J = 7.6 Hz, 4H), 7.61 (d, J = 9.2 Hz, 1H), 7.68-7.72 (m, 4H), 7.95 (s, 1H), 8.16 (d, J = 8.8 Hz, 1H), 8.45 (d, J = 5.6 Hz, 4H). Mass: (EM+2H+)/2: found 510.
Compound 4; 1H NMR (300 MHz, CDC13) a 1.05 (t, J = 7.2 Hz, 3H), 1.39 (t, J = 7.5 Hz, 3H), 1.97-1.83 (m, 6H), 3.13 (q, J = 7.5 Hz, 2H), 3.50 (s, 4H), 3.54 (s, 8H), .64 (m,, 2H), 4.04 (m, 2H), 5.28 (d, J = 11.7 Hz, 2H), 5.30 (d, J = 16.8 Hz, 1H), 5.74 (d, J = 16.2 Hz, 1H), 6.80 (s, 2H), 7.12 (s, 1H), 7.31-7.18 (m, 12H), 738—7.41 (m, 8H), 7.55-7.60 (m, 1H), 7.66-7.68 (m, 2H), 7.93 (d, J = 2.1 Hz, 1H), 8.06 (d, J = 7.8 Hz, 1H), 8.27 (d, J = 9 Hz, 1H), 8.34 (d, J = 7.5 Hz, 1H), 8.63 (s, 1H). Mass: (EM+2H+)/2: found 553.74. nd 5; 1H NMR (300 MHz, CDC13) o 1.05 (t, J = 7.2 Hz, 3H), 1.41 (t, J = 7.8 HZ, 3H), 1.83-1.97 (m, 6H), 2.78 (t, J = 7.2 HZ, 2H), 3.13-3.21 (m, 2H), 3.65 (S, 4H), 3.80 (S, 8H), 3.96-4.00 (m, 4H), 5.27 (s, 2H), 5.31 (d, J = 16.2 Hz, 1H), 5.76 (d, J = 16.2 Hz, 1H), 6.84 (s, 2H), 7.07 (s, 10-7.15(n1,4H),7.52-7.66(n1, 11H), 7.69 (d, J = 2.1 Hz, 1H), 7.95 (d, J = 1.8 HZ, 1H), 8.21 (d, J = 7.8 HZ, 2H), 8.28 (d, J = 9 HZ, 1H), 8.50 (d, J = 4.8 HZ, 4H). Mass: +)/2: found 549.
Compound 6: 1H NMR (300 MHz, CDC13): a = 8.61 (t, J = 1.5 Hz, 1H), 8.49 (dt, J = 4.8, 1.2 Hz, 4H), 8.34 (dt, J = 7.8, 1.5 Hz, 1H), 8.25 (d, J = 9.0 Hz, 1H), 8.07 (dt, J = 8.1, 1.5 Hz, 1H), 7.92 (d, J = 2.1 Hz, 1H), 7.67-7.63 (m, 3H), 7.62-7.56 (m, 9H), 7.41 (d, J = 7.5 Hz, 1H), 7.30—7.27 (m, 5H), 7.25-7.18 (m, 1H), 7.14-7.10(n1, 4H), 7.07 (s, 1H), 6.80 (s, 2H), 6.15-6.11 (m, 1H), 5.73 (d, J = 16.5 Hz, 1H), 5.31—5.25 (m, 3H), 4.84 (dd, J = 14.0, 8.0 Hz, 1H), 3.89 (t, J = 5.6 Hz, 2H), 3.79 (s, 8H), 3.64 (s, 4H), 3.36-3.10 (m, 6H), 1.97-1.83 (m, 2H), 1.66-1.57 (m, 4H), 1.40 (t, J = 7.6 Hz, 3H), 1.04 (t, J = 7.4 Hz, 3H). Mass: (EM+2H+)/2: found 629.
Compound 7; 1H NMR (300 MHz, CDC13): a = 8.46 (d, J = 4.5 Hz, 4H), 8.28 (d, J = 8.4 Hz, 2H), 8.24 (d, J = 9.3 Hz, 1H), 7.97 (d, J = 8.4 Hz, 2H), 7.87 (s, 1H), .55 (m, 10H), 7.11—7.07 (m, 4H), 7.04 (s, 1H), 6,79 (s, 2H), 5.68 (d, J = 16.5 Hz, 1H), 5.27—5.22 (m, 3H), 3.95-3.89 (m, 2H), 3.77 (s, 8H), 3.62 (s, 4H), 3.10-3.08 (m, 4H), .72 (m, 6H), 1.35 (t, J = 7.5 Hz, 3H), 0.98 (t, J = 7.2 Hz, 3H). Mass: +)/2; found 574.
Compound 8: 1H NMR (400 MHz, CDC13): a = 8.48 (d, J = 4.4 Hz, 4H), 8.38 (s, 1H), 8.04-8.00 (m, 2H), 7.76-7.71 (m, 3H), 7.60-7.57 (m, 4H), 7.54—7.51 (m, 5H), 7.48 (dd, J = 9.2, 1.6 Hz, 1H), 7.30 (t, J = 8.0 Hz, 1H), 7.11-7.08 (m, 4H), 6.96 (s, 1H), 6.83 (s, 2H), 6.06 (m, 1H), 5.68 (d, J = 16.0 Hz, 1H), 5.22 (d, J = 16.0 Hz, 1H), 5.19 (d, J = 27.2 Hz, 1H), 5.14 (d, J = 27.2 Hz, 1H), 3.99 (t, J = 6.0 Hz, 2H), 3.76 (s, 8H), 3.60 (s, 4H), .36 (m, 2H), 3.00 (q, J = 7.2 Hz, 2H), 1.92-1.81 (m, 4H), 1.74 (q, J = 6.8 Hz, 2H), 1.32 (t, J = 7.6 Hz, 3H), 0.99 (t, J = 7.2 Hz, 3H). Mass: (EM+2H+)/2; found 563.25.
Compound 9; 1H NMR (400 MHz, CDC13): a = 8.65 (s, 1H), 8.49 (d, J = 4.4 Hz, 4H), 8.41 (s, 1H), 8.26 (d, J = 8 Hz, 1H), 8.16-8.08 (m, 3H), 8.01 (d, J = 8Hz, 1H), 7.87 (d, J = 2.4 Hz, 2H), 7.64-7.51 (m, 8H), 7.14 (t, J = 6 Hz, 4H), 7.00 (s, 1H), 6.79 (d, J = 4.4 Hz, 2H), 5.45 (dd, J = 172, 16.4, 2H), 5.21 (s, 1H), 5.08 (s, 2H), 5.03 (s, 1H), 3.88-3.82 (m, 10H), 3.66 (s, 4H), 3.58-3.46 (m, 4H), 3.25—3.22 (m, 2H), 3.10—2.95 (m, 4H), 2.58-2.51 (m, 2H), 1.94-1.83 (m, 2H), 1.70—1.57 (m, 4H), 1.36 (t, J = 8 Hz, 3H), 1.02 (t, J = 8 Hz, 3H). Mass: (EM+2H+)/2; found 667.29.
Compound 11; 1H NMR (300 MHz, CDC13): a = 8.47 (dd, J = 5.1, 0.9 Hz, 4H), 8.22- 8.19 (m, 2H), 7.85-7.82 (m, 2H), 7.64-7.52 (m, 16H), 7.43—7.30 (m, 6H), 7.11—7.04 (m, 5H), 6.85 (s, 2H), 5.71 (d, J = 16.2 Hz, 1H), 5.27 (d, J = 16.2 Hz, 1H), 5.22 (s, 2H), 4.68 (s, 2H), 4.05 (m, 2H), 3.76 (s, 8H), 3.61 (4H), 3.57 (m, 2H), 3.09 (q, J = 7.5 Hz, 2H), 1.93-1.83 (m, 6H), 1.35 (t, J = 7.8 Hz, 3H), 1.02 (t, J = 7.5 Hz, 3H). Mass: (EM+2H+)/2; found 646.29.
Compound 12; 1H NMR (300 MHz, CDC13) a 1.03 (t, J = 7.5 Hz, 3H), 1.39 (t, J = 7.5 Hz, 3H), 1.85 (m, 6H), 2.58-2.51 (m, 1H), 3.24—3.03 (m, 6H), .29 (m, 1H), 3.52-3.48 (m, 2H), 3.67 (s, 4H), 3.82 (s, 8H), 3.91-3.87 (m, 1H), 4.00 (t, J = 4.8 Hz, 2H), 4.69 (d, J = 3.0 Hz, 2H), 5.27 (d, J = 12.0 Hz, 2H), 5.30 (d, J = 16.5 Hz, 1H), 5.74 (d, J =16.2 Hz, 1H), 6.86 (s, 2H), 7.04 (s, 1H),7.15-7.11 (m, 4H), .36 (m, 2H), 7.71—7.55 (m, 12H), 7.83 (d, J = 2.1 Hz, 1H), 8.23 (d, J = 9.3 Hz, 1H), 8.49 (d, J = 4.8 Hz, 4H). Mass: (EM+2H+)/2: found 641.26.
Compound 13; 1H NMR (300 MHz, CDC13) a 1.06-0.88 (m, 6H), 1.40 (t, J = 7.5 Hz, 3H), 1.83-1.65 (m, 4H), 1.97-1.85 (m, 2H), 2.26 (t, J = 7.5 Hz, 1H), 2.41 (t, J = 7.2 Hz, 1H), 3.16 (m, 2H), 3.35 (m, 1H), 3.49 (m, 1H), 3.65 (s, 2H), 3.67 (s, 2H), 3.80 (s, 8H), 3.96 (m, 2H), 4.68 (s, 1H), 4.72 (s, 1H), 5.27 (s, 2H), 5.31 (d, J = 14.1 Hz, 1H), 5.75 (d, J = 16.2 Hz, 1H), 6.84 (s, 2H), 7.15—7.07 (m, 5H), 7.41—7.34 (m, 2H), 7.69-7.56 (m, 10H), 7.96-7.94 (m, 1H), 8.18 (d, J = 8.4 Hz, 1H), 8.31-8.24 (m, 2H), 8.50 (d, J = 4.5 Hz, 4H). Mass: (EM+2H+)/2: found 583.27.
Compound 14: 1H NMR (300 MHz, CDC13) 8 1.04 (t, J = 7.5 Hz, 3H), 1.40 (t, J = 8.1 Hz, 3H), 1.97-1.81 (m, 6H), 3.21-3.13 (m, 2H), 3.60 (t, J = 7.2 Hz, 2H), 3.67 (s, 4H), 3.82 (s, 8H), 3.97 (m, 2H), 4.92 (s, 2H), 5.28 (d, J = 3.9 Hz, 2H), 5.32 (d, J = 13.2 Hz, 1H), 5.76 (d, J = 16.2 Hz, 1H), 6.85 (s, 2H), 6.95-6.92 (m, 1H), 7.15-7.10 (m, 5H), 7.41 (d, J = 4.8 Hz, 1H), 7.48 (d, J = 7.8 Hz, 2H), .56 (m, 11H), 7.96 (m, 1H), 8.31-8.24 (m, 3H), 8.50 (d, J = 4.5 Hz, 4H). Mass: (EM+2H+)/2: found 603.74.
Compound 15; 1H NMR (300 MHz, CDC13): a = 8.51 (d, J = 4.2 Hz, 4H), .16 (m, 3H), 7.98-7.95 (m, 1H), 7.72—7.53 (m, 10H), 7.37 (d, J = 8.1Hz, 2H), 7.18-7.11 (m, 4H), 7.08 (s, 1H), 6.84 (s, 2H), 5.57 (dd, J = 133, 16.5Hz, 2H), 5.28 (s, 2H), 4.69 (d, J = 4.8 Hz, 2H), 3.97 (d, J = 5.4 Hz, 2H), 3.86-3.72 (m, 10H), 3.70-3.56 (m, 4H), 3.52—3.29 (m, 2H), 3.22-3.13(m,2H),1.96-1.84(m,2H), 1.82-1.54 (m, 12H), 1.41 (t, J = 7.2 Hz, 3H), 1.30—1.14 (m, 3H), 1.05 (t, J = 7.2 Hz, 3H). Mass: (EM+2H+)/2; found 603.78.
Compound 16: 1H NMR (400 MHz, CDC13) a 1.05 (t, J = 7.2 Hz, 3H), 1.41 (t, J = 7.6 Hz, 3H), 1.96-1.85 (m, 4H), 3.17 (q, J = 8.0 Hz, 2H), 3.29 (s, 1H), 3.57 (s, 1H), 3.69 (s, 4H), 3.83 (s, 9H), 4.00 (s, 1H), 4.64 (s, 1H), 4.87 (s, 1H), 5.29 (d, J = 5.6 Hz, 2H), 5.32 (d, J = 17.2 Hz, 1H), 5.76 (d, J = 16 Hz, 1H), 6.81 (s, 1H), 6.86 (s, 1H), 7.15-7.12(m, 5H), 7.31— 7.29 (m, 2H), 7.49 (m, 3H), .57 (m, 9H), 7.69-7.66 (m, 2H), 7.96 (d, J = 2.8 Hz, 1H), 8.25 (d, J = 8.4 Hz, 2H), 8.30 (d, J = 9.2 Hz, 1H), 8.50 (d, J = 4.8 Hz, 4H). Mass: (EM+2H+)/2: found 639.72. nd 17: 1H NMR (400 MHz, CDC13) 8 1.00 (t, J = 7.2 Hz, 3H), 1.38 (t, J = 7.6 Hz, 3H), 1.63 (m, 1H), 1.81-1.94 (m, 5H), 3.13 (q, J = 7.2 Hz, 2H), 3.37 (m, 1H), 3.61 (m, 5H), 3.78 (m, 9H), 4.00 (m, 1H), 4.71 (s, 1H), 4.90 (s, 1H), 5.24 (d, J = 8 Hz, 2H), 5.27 (d, J = 16 Hz, 1H), 5.70 (d, J = 16.4 Hz, 1H), 6.77 (s, 1H), 6.84 (s, 1H), 7.07-7.11 (m, 5H), 7.32- 7.38 (m, 4H), 7.50-7.56 (m, 15H), 7.65 (d, J = 9.2 Hz, 1H), 7.68 (s, 1H), 7.93 (d, J = 2.4 Hz, 1H), 8.23 (d, J = 8.4 Hz, 2H), 8.27 (d, J = 9.2 Hz, 1H), 8.47 (d, J = 4.4 Hz, 4H). Mass: (EM+2H+)/2: found 638.83.
Compound 18: 1H NMR (300 MHZ, CDC13) 8 1.04 (t, J = 7.5 Hz, 3H), 1.37 (t, J = 7.8 Hz, 3H), 1.50-1.46 (m, 2H), 1.77 (m, 2H), 1.93-1.83 (m, 2H), 3.16 (q, J = 7.5 Hz, 2H), 3.66 (s, 4H), 3.74-3.72 (m, 2H), 3.81 (s, 8H), 4.09-3.96 (m, 4H), 4.82 (d, J = 9.3 Hz, 2H), 5.28 (d, J = 4.2 Hz, 2H), 5.31 (d, J = 14.7 Hz, 1H), 5.75 (d, J = 16.2 Hz, 1H), 6.75 (s, 1H), 6.85 (s, 1H), 7.26-7.10 (m, 15H), 7.50 (d, J = 7.8 Hz, 2H), 7.68-7.55 (m, 10H), 7.95 (d, J = 1.8 Hz, 1H), 8.21-8.17 (m, 2H), 8.29 (d, J = 9.0Hz, 1H), 8.49 (d, J = 4.5 Hz, 4H). Mass: (EM+2H+)/2: found 645.79.
Compound 19; 1H NMR (300 MHz, CDC13) a 1.01 (t, J = 7.2 Hz, 3H), 1.38 (t, J = 7.5 HZ, 3H), 1.74 (m, 2H), 1.93-1.84 (m, 6H), 3.18-3.10 (m, 2H), 3.39-3.44 (m, 2H), 3.64 (S, 4H), 3.78 (s, 8H), 3.94 (t, J = 5.7 Hz, 2H), 4.62 (s, 2H), 5.31—5.25 (m, 3H), 5.71 (d, J = 16.5 HZ, 1H), 6.81 (S, 2H), 7.13-7.08 (m, 6H), 7.43 (d, J = 8.7 HZ, 2H), 7.62-7.54 (m, 8H), 7.65 (d, J = 2.4 Hz, 1H), 7.67 (s, 1H), 7.92 (d, J = 2.4 Hz, 1H), 8.15 (d, J = 2.7 Hz, 1H), 8.20 (d, J = 8.1 Hz, 2H), 8.27 (d, J = 9.6 Hz, 1H), 8.48 (d, J = 4.8 Hz, 4H), 8.64 (d, J = 2.7 Hz, 1H).
Mass: (EM+2H+)/2: found 632.
Compound 20; 1H NMR (300 MHz, CDC13) a 1.04 (t, J = 7.8 Hz, 3H), 1.40 (t, J = 7.2 HZ, 3H), 1.77 (m, 2H), 1.95-1.85 (m, 4H), 3.17-3.12 (m, 2H), 3.56 (m, 2H), 3.69 (S, 4H), 3.82 (S, 8H), 3.96 (m, 2H), 4.92 (S, 2H), 5.28 (S, 2H), 5.30 (d, J = 16.5 HZ, 1H), 5.74 (d, J = 16.5 Hz, 1H), 6.82 (s, 2H), 7.08 (s, 1H), .12 (m, 4H), 7.42 (d, J = 7.8 Hz, 2H), 7.67-7.56 (m, 10H), 7.95 (s, 1H), 8.23 (d, J = 7.5 Hz, 2H), 8.28 (d, J = 9 Hz, 1H), 8.52 (m, 4H), 8.68 (s, 1H). Mass: (EM+2H+)/2: found 627. nd 21: 1H NMR (300 MHz, CDC13) 8 1.03 (t, J = 7.2 Hz, 3H), 1.39 (t, J = 7.5 Hz, 3H), 1.96-1.85 (m, 4H), 2.09 (m, 2H), 3.16-3.11 (m, 2H), 3.64 (m, 2H), 3.67 (s, 4H), 3.80 (s, 8H), 4.06 (t, J = 6 Hz, 2H), 4.13 (m, 2H), 5.33—5.27 (m, 3H), 5.74 (d, J = 16.8 Hz, 1H), 6.17 (d, J = 8.4 Hz, 1H), 6.84 (s, 2H), 7.15—7.10 (m, 5H), 7.39 (d, J = 8.1 Hz, 2H), 7.66-7.54 (m, 10H), 7.94 (s, 1H), 8.28-8.23 (m, 3H), 8.35 (d, J = 9.3 Hz, 1H), 8.48 (d, J = 5.1 Hz, 4H).
Mass: (EM+2H+)/2: found 630.
Compound 22; 1H NMR (400 MHz, CDC13): a = 8.51 (s, 4H), 8.31-8.18 (m, 3H), 7.97—7.94 (m, 1H), 7.72-7.60 (m, 10H), 7.44—7.30 (m, 2H), 7.19—7.07 (m, 5H), 6.86 (s, 2H), .54 (dd, J = 172, 16.4Hz, 2H), 5.28 (s, 2H), 4.73 (d, J = 16.4 Hz, 2H), 3.99 (s, 2H), 3.83- 3.74 (m, 10H), .67 (m, 4H), 3.50 (s, 1H), 3.38 (s, 1H), 3.18 (s, 2H), 2.50—2.42 (m, 2H), 1.97-1.86 (m, 2H), 1.81 (s, 4H), 1.73-1.67 (m, 2H), 1.45—1.39 (m, 3H), .30 (m, 6H), 1.08-1.02 (m, 3H), .87 (m, 3H). Mass: (EM+2H+)/2: found 604.79.
Compound 23: 1H NMR (400 MHZ, CDC13) 5 1.04 (t, J = 7.2 HZ, 3H), 1.41 (t, J = 7.6 HZ, 3H), 1.59 (m, 1H), 1.76 (m, 1H), 1.96-1.83 (m, 4H), 3.20-3.14 (m, 2H), 3.30 (m, 1H), 3.59 (m, 1H), 3.67 (S, 4H), 3.81 (S, 8H), 3.89-4.02 (m, 2H), 4.66 (S, 1H), 4.90 (S, 1H), 5.28 (d, J = 5.6 HZ, 2H), 5.31 (d, J = 17.2 HZ, 1H), 5.75 (d, J =16.4 HZ, 1H), 6.78 (S, 1H), 6.86 (S, 1H), 7.14-7.10 (m, 5H), 7.42-7.35 (m, 5H), 7.63-7.57 (m, 10H), 7.69-7.66 (m, 2H), 7.96 (d, J = 2.8 Hz, 1H), 8.24 (d, J = 8 Hz, 2H), 8.29 (d, J = 9.2 Hz, 1H), 8.50 (d, J = 4.8 Hz, 4H).
Mass: (EM+2H+)/2: found 600.76.
Compound 24: 1H NMR (300 MHz, CDC13) 8 1.04 (t, J = 7.5 Hz, 3H), 1.44-1.37 (m, 4H), 1.70-1.65 (m, 1H), 1.99-1.86 (m, 4H), 3.19-3.15 (m, 2H), 3.68-3.65 (m, 6H), 3.82-3.80 (m, 10H), 4.09-3.94 (m, 2H), 5.29 (s, 2H), 5.31 (d, J = 16.2 Hz, 1H), 5.75 (d, J = 16.2 Hz, 1H), 6.68 (s, 1H), 6.90 (s, 1H), 7.14-7.12 (m, 5H), 7.28(d, J = 5.1 Hz, 2H), 7.64-7.39 (m, 12H), 7.72-7.67 (m, 2H), 7.99-7.80 (m, 4H), 8.17 (d, J = 8.1 Hz, 1H), 8.32-8.27 (m, 2H), 8.50 (d, J = 4.5 Hz, 4H). Mass: (EM+2H+)/2: found 625.77.
Compound 25; 1H NMR (300 MHz, CDC13) a 1.00 (t, J = 7.2 Hz, 3H), 1.38 (t, J = 7.5 Hz, 3H), 1.60-1.49 (m, 4H), 1.93-1.81 (m, 2H), 2.46 (s, 1H), 2.54 (s, 2H), 3.03 (m, 1H), 3.17— 3.10 (m, 2H), 3.44 (m, 1H), 3.64 (s, 4H), 3.78 (s, 8H), 3.86 (m, 2H), 4.50 (s, 1H), 4.72 (s, 1H), 5.25 (d, J = 4.2 Hz, 2H), 5.27 (d, J = 16.8 Hz, 1H), 5.70 (d, J = 16.5 Hz, 1H), 6.80-6.77 (m, 2H), 7.12-7.08 (m, 5H),7.19-7.17 (m, 1H), 7.35—7.33 (m, 1H), .42 (m, 3H), 7.65- 7.54 (m, 11H), 7.68 (s, 1H), 7.92 (d, J = 2.4 Hz, 1H), .06 (m, 2H), 8.28 (d, J = 9.0 Hz, 1H), 8.47 (d, J = 4.8 Hz, 4H). Mass: (EM+2H+)/2: found 658.26.
Compound 26: 1H NMR (300 MHz, CDC13) a 1.04 (t, J = 7.2 Hz, 3H), 1.29 (m, 2H), 1.40 (t, J = 7.5 Hz, 3H), 1.68 (m, 8H), 1.95-1.86 (m, 4H), 2.28 (m, 2H), 2.95 (m, 1H), 3.16 (q, J = 7.5 HZ, 2H), 3.30 (m, 1H), 3.67 (S, 4H), 3.81 (S, 8H), 3.91 (m, 2H), 4.22 (S, 1H), 4.65 (s, 1H), 5.28 (d, J = 3.6 Hz, 2H), 5.31 (d, J =15.3 Hz, 1H), 5.76 (d, J = 16.5 Hz, 1H), 6.81 (s, 2H), 7.15—7.10 (m, 5H), 7.38-7.20 (m, 6H), 7.61-7.56 (m, 8H), 7.69-7.64 (m, 2H), 7.95 (d, J = 1.8 Hz, 1H), 8.17 (d, J = 7.8 Hz, 2H), 8.29 (d, J = 9.3 Hz, 1H), 8.50 (d, J = 4.2 Hz, 4H).
Mass: (EM+2H+)/2: found 658.28.
Compound 27: 1H NMR (300 MHz, CDC13) 8 1.04 (t, J = 7.2 Hz, 3H), 1.41 (t, J = 7.5 Hz, 3H), 1.77-1.60 (m, 2H), 1.93-1.84 (m, 4H), 3.17 (q, J = 7.2 Hz, 2H), 3.27 (m, 1H), 3.61 (m, 1H), 3.66 (s, 4H), 3.81 (s, 8H), 4.01-3.94 (m, 2H), 4.61 (s, 1H), 4.90 (s, 1H), 5.28 (s, 2H), .31 (d, J = 15.3 Hz, 1H), 5.75 (d, J = 16.2 Hz, 1H), 6.79 (s, 1H), 6.85 (s, 1H), 7.14-7.10 (m, 5H), .55 (m, 16H), 7.96 (s, 1H), 8.25 (d, J = 7.5 Hz, 2H), 8.30 (d, J = 9.0 Hz, 1H), 8.50 (d, J = 4.8 Hz, 4H). Mass: (EM+2H+)/2: found 634.76.
Compound 28: 1H NMR (300 MHZ, CDC13): 5 = 8.49 (dd, J = 4.5, 0.9 HZ, 4H), 8.16 (d, J = 9.3 HZ, 1H), 7.78 (d, J = 2.1 HZ, 1H), 7.63-7.50 (m, 10H), 7.13-7.09 (m, 4H), 7.06 (S, 1H), 6.85 (S, 2H), 5.85 (t, J = 6.0 HZ, 1H), 5.72 (d, J = 16.2 HZ, 1H), 5.28 (d, J =16.2 HZ, 1H), 5.17 (S, 2H), 4.01 (t, J = 5.4 HZ, 2H), 3.80 (S, 8H), 3.65 (S, 4H), 3.40 (q, J = 6.0 HZ, 2H), 3.07 (q, J = 7.5 HZ, 2H), 1.95-1.82 (m, 6H), 1.34 (t, J = 7.5 HZ, 3H), 1.01 (t, J = 7.2 HZ, 3H).
Mass: (EM+2H+)/2: found 504. nd 29; 1H NMR (300 MHz, CDC13): a = 8.48 (d, J = 4.8 Hz, 4H), 8.19 (d, J = 9.0 Hz, 1H), 7.85-7.55 (m, 12H), 7.12-7.08 (m, 5H), 6.79 (s, 2H), 5.71 (dd, J = 16.5, 7.2 Hz, 1H), 5.29 (d, J = 16.5 Hz, 1H), 5.21 (d, J = 7.8 Hz, 2H), 3.92 (m, 1H), .74 (m, 11H), 3.63-3.59 (m, 5H), 3.36-3.34 (m, 2H), 3.11-3.08 (m, 2H), 2.40—1.72 (m, 10H), .32 (m, 3H), 1.04 (t, J = 7.2 Hz, 3H). Mass: (EM+2H+)/2: found 552.
Compound 31; 1H NMR (400 MHz, CDC13): a = 8.49 (d, J = 4.4 Hz, 4H), 8.23 (dd, J = 9.2, 5.2 Hz, 1H), 7.88-7.56 (m, 15H), 7.45—7.41 (m, 4H), 7.35 (t, J = 6.4 Hz, 1H), 7.11-7.08 (m, 5H), 6.84 (s, 1H), 6.79 (s, 1H), 5.73 (d, J = 16.0 Hz, 1H), 5.30 (d, J = 16.0 Hz, 1H), 5.24 (s, 2H), 4.79 (s, 1H), 4.68 (s, 1H), 3.99 (t, J = 5.6 Hz, 2H), 3.80-3.77 (m, 8H), 3.64-3.61 (m, 4H), 3.58-3.54 (m, 2H), 3.12-3.10 (m, 2H), 1.89-1.88 (m, 6H), 1.38-1.33 (m, 3H), 1.04-1.01 (m, 3H). Mass: (EM+2H+)/2: found 586.77.
Compound 32: 1H NMR (400 MHz, CDC13): 8 = 8.48 (d, J = 4.8 Hz, 4H), 8.15 (t, J :80 Hz, 1H), 7.77 (s, 1H), 7.62-7.54 (m, 10H), 7.11 (t, J = 6.0 Hz, 4H), 7.05 (s, 1H), 6.82 (s, 2H), 6.13-6.09 (m, 1H), 5.69 (d, J = 16.4 Hz, 1H), 5.26 (d, J = 16.4 Hz, 1H), 5.20 (s, 2H), 4.36 (d, J = 12.8 Hz, 1H), 4.25 (d, J = 12.0 Hz, 1H), 3.96 (t, J = 5.6 Hz, 2H), 3.78 (s, 8H), 3.63 (s, 4H), 3.34 (q, J = 6.4 Hz, 2H), 3.13-3.04 (m, 3H), 2.87 (t, J = 11.2 Hz, 1H), .28 (m, 1H), 1.93-1.79 (m, 8H), 1.71 (q, J = 7.6 Hz, 2H), 1.37 (t, J = 7.6 Hz, 3H), 0.99 (t, J = 7.2 Hz, 3H). Mass: (EM+2H+)/2: found 559.26. nd 33; 1H NMR (300 MHz, CDC13): a = 8.47 (d, J = 4.8 Hz, 4H), 8.20 (d, J = 9.0 Hz, 1H), 7.80-7.78 (m, 1H), 7.64-7.49 (m, 10H),7.47-7.41 (m, 1H), 7.10 (t, J = 6.0 Hz, 4H), 7.04 (s, 1H), 6.78 (s, 2H), 5.69 (d, J = 16.5 Hz, 1H), 5.27 (d, J = 16.5 Hz, 1H), 5.22 (s, 2H), 3.91 (m, 2H), 3.82 (m, 2H), 3.76 (s, 8H), .68 (m, 4H), 3.61 (s, 6H), 3.31—3.27 (m, 2H), 3.11 (q, J = 7.2 Hz, 2H), 2.47—2.44 (m, 2H), 2.35—2.33 (m, 2H), 2.05—1.95 (m, 2H), 1.92— 1.83 (m, 2H), 1.81-1.76 (m, 2H), 1.66 (m, 2H), 1.36 (t, J = 7.5 Hz, 3H), 1.12—1.09 (m, 6H), 0.99 (t, J = 7.5 Hz, 3H). Mass: (EM+2H+)/2: found 615.80.
Compound 34: 1H NMR (400 MHz, CDC13): 8 = 8.38 (d, J = 4.0 Hz, 4H), 7.74-7.71 (m, 4H), 7.62-7.58 (m, 7H), 7.35 (dd, J = 7.2, 2.4 Hz, 1H), 7.24-7.21 (m, 4H), 6.89 (s, 1H), 6.58 (s, 2H), 5.35 (m, 1H), 4.97 (m, 1H), 4.73 (s, 2H), 4.25-4.24 (m, 1H), 4.13-4.10 (m, 1H), 3.91 (s, 3H), 3.76 (t, J = 5.6 Hz, 2H), 3.68 (s, 8H), 3.61 (m, 1H), 3.49 (s, 4H), 3.20-3.17 (m, 1H), 3.08-3.03 (m, 1H), 2.94 (d, J = 18.4 Hz, 1H), 2.80 (d, J = 18.4 Hz, 1H), 2.49-2.30 (m, 6H), .93 (m, 2H), 1.71-1.53 (m, 4H), 1.25 (d, J = 6.8 Hz, 3H). Mass: (EM+2H+)/2: found 605. nd 35: 1H NMR (400 MHz, CDC13): 8 = 8.49 (d, J = 4 Hz, 4H), 8.18 (d, J = 9.2Hz, 1H), 7.79 (s, 1H), 7.68-7.41 (m, 11H), 7.15-7.06 (m, 6H), 5.52 (dd, J = 175.6, 16.4Hz, 2H), 5.22 (s, 2H), 4.14 (s, 2H), 3.96 (s, 4H), 3.83-3.73 (m, 8H), 3.48 (s, 2H), 3.09-3.07 (m, 2H), 1.94-1.86 (m, 2H), 1.34 (t, J = 7.6Hz, 3H), 1.019 (t, J = 7.6Hz, 3H). Mass: (EM+2H+)/2: found 510.
WO 79299 Compound 36: 1H NMR (400 MHz, CDC13): a = 8.68 (s, 1H), 8.49 (d, J = 4.4 Hz, 4H), 8.24 (t, J = 8Hz, 2H), 8.09 (d, J = 8 Hz, 1H), 7.87 (d, J = 2Hz, 1H), 7.67 (s, 1H), 7.58- 7.43 (m, 10H), 7.11—7.04 (m, 6H), 5.54 (dd, J = 178.6, 16Hz, 2H), 5.27 (s, 2H), .76 (m, 11H), 3.49 (s, 3H), 3.15 (q, J = 7.6Hz, 2H), 2.90 (t, J = 6.4 Hz, 2H), 1.95-1.84 (m, 2H), 1.38 (t, J = 7.6Hz, 3H), 1.06 (t, J = 7.6Hz, 3H). Mass: (EM+2H+)/2; found 542.
Compound 37; 1H NMR (300 MHz, CDC13): a = 8.50-8.48 (m, 4H), 8.31-8.24 (m, 3H), 7.97 (S, 1H), 7.70-7.52(n1, 12H), 7.16-7.10 (In, 6H), 7.06 (S, 1H), 7.04 (S, 1H), 6.78 (S, 2H), 6.05 (S, 1H), 5.76 (d, J = 16.5 HZ, 1H), 5.34-5.28 (In, 3H), 4.74 (S, 2H), 4.04 (In, 2H), 3.77 (s, 8H), 3.59 (s, 4H), 3.55 (m, 2H), 3.17 (q, J = 7.5 Hz, 2H), 2.54 (q, J = 7.5 Hz, 4H), 1.97-1.84 (In, 6H), 1.41 (t, J = 7.8 HZ, 3H), 1.12 (t, J = 7.5 HZ, 6H), 1.04 (t, J = 7.2 HZ, 3H).
Mass: (EM+2H+)/2: found 636.
Compound 38: 1H NMR (300 MHZ, CDC13) 5 1.04 (t, J = 7.5 HZ, 3H), .25 (m, 2H), 1.40 (t, J = 7.5 HZ, 3H), 1.63 (m, 6H), 1.93-1.85 (m, 6H), 2.22-2.17 (m, 2H), 2.90 (m, 1H), 3.20-3.15 (m, 3H), 3.88-3.67 (m, 26H), 4.09 (S, 1H), 4.54 (S, 1H), 5.28 (d, J = 10.5 HZ, 2H), 5.31 (d, J = 12.0 HZ, 1H), 5.75 (d, J = 16.2 HZ, 1H), 6.77 (S, 2H), 6.97 (m, 1H), 7.26- 7.11 (m, 10H), 7.72-7.55 (m, 12H), 7.99-7.96 (m, 3H), 8.29-8.26 (m, 3H), 8.49 (d, J = 4.8 Hz, 4H). Mass: (EM+2H+)/2: found 798.
Compound 39; 1H NMR (400 MHz, CDC13): a = 1.03 (t, J = 7.2 Hz, 3H), 1.25 (m, 1H), 1.39 (t, J = 7.6 Hz, 3H), 1.65-1.96 (m, 11H), 2.01 (m, 2H), 2.24 (m, 2H), 3.13—3.19 (m, 3H), 3.44—3.54 (m, 5H), 3.59-3.64 (m, 12H), 3.77 (s, 8H), 3.97 (t, J = 5.6 Hz, 2H), 4.30 (s, 1H), 4.67 (s, 1H), 5.27 (d, J =12.4 Hz, 2H), 5.30 (d, J = 16.0 Hz, 1H), 5.74 (d, J = 16.0 Hz, 1H), 6.80 (s, 2H), 7.05 (s, 1H), 7.09-7.12(n1, 5H), 7.20—7.30 (m, 5H), 7.54-7.62 (m, 8H), 7.65-7.68 (m, 2H), 7.76 (m, 4H), 7.94 (d, J = 2.4 Hz, 1H), 8.13 (d, J = 6.8 Hz, 2H), 8.28 (d, J = 9.2 Hz, 1H), 8.48 (d, J = 4.8 Hz, 4H). Mass: (EM+2H+)/2; found 798.
Compound 40: 1H NMR (300 MHz, CDC13): 8 = 1.04 (t, J = 7.2 Hz, 3H), 1.25-1.29 (m, 1H), 1.39 (t, J = 7.2 Hz, 3H), .97 (m, 9H), 2.17-2.26 (m, 3H), 2.75 (m, 1H), 3.12- 3.20 (m, 3H), 3.39 (m, 1H), 3.74-3.86 (m, 12H), 4.10 (s, 1H), 4.72 (s, 1H), 5.27-5.34 (m, 3H), 5.75 (d, J = 16.5 Hz, 1H), 6.99 (s, 2H), 7.12-7.14 (m, 4H), 7.20-7.31 (m, 6H), .46 (m, 4H), 7.55-7.60 (m, 4H), 7.64-7.67 (m, 3H), 7.93 (s, 1H), 8.10-8.18 (m, 2H), 8.28 (d, J = 9.3 Hz, 1H), 8.51 (m, 4H). Mass: (EM+2H+)/2: found 645.
Compound 41; 1H NMR (CDC13, 400 MHz): 3 = 1.12 (s, 3H), 1.21 (s, 3H), 1.68 (s, 3H), 1.91 (s, 3H), 1.63-2.33 (m, 4H), 2.26 (s, 3H), 2.41 (s, 3H), 2.38-2.58 (m, 8H), 2.72 (t, J = 6.8 Hz, 2H), .27 (m, 2H), 3.64(s, 4H), 3.78(s, 8H), 3.940, J = 6.0 Hz, 2H), 4.19(d, J = 8.4 Hz, 1H), 4.30(d, J = 8.4 Hz, 1H), 4.43(dd, J = 10.8, 6.8 Hz, 1H), 4.96(d, J = 9.2 Hz, 1H), , J = 7.2 Hz, 1H), 5.88-5.93(m, 2H), 6.20(t, J = 9.2 Hz, 1H), 6.29(s, 1H), 6.81(s, 2H), 7.06 (s, 1H), 7.06-7.12(m, 4H), 7.27-7.63(m, 24H), 7.79(d, J = 5.6 Hz, 2H), 8.13 (d, J = .6 Hz, 2H), 8.48(d, J = 4.8 Hz, 4H). Mass: (EM+2H+)/2: found 762.
Compound 42: 1H NMR (CDC13, 400 MHz): 8 = 0.98(t, J = 7.2 Hz, 3H), 1.22(m,8H), 1.38(t, J = 7.6 Hz, 3H), 1.88(m, 2H), 2.01(s, 5H), 2.10-2.41(m, 8H), 2.85(s, 1H), 2.92(s, 1H), 3.15(m, 3H), 3.44(s, 2H), .71(m, 13H), 3.76(s, 8H), 3.86(s, 2H), 4.10(dd, J = 7.2 Hz, 4H), 4.32(s, 2H), 4.53(s, 1H), 5.24(s, 2H), 5.30(d, J = 16 Hz, 1H), 5.73(d, J = 16 Hz, 1H), 6.75(s, 3H), 6.95(br, 2H), 7.1-7.25(m, 13H), , J = 8.4 Hz, 2H ), 7.52-7.64(m, 8H), 7.64- 7.72(m, 3H), 7.93(d, J = 2.4 Hz, 1H), 8.14(d, J = 7.2 Hz, 2H), 8.27(d, J = 9.2 Hz, 1H), 8.47 (d, J = 8.8 Hz, 4H). Mass: (EM+2H+)/2: found 901.6.
Compound 43; 1H NMR (CDC13, 400 MHz): 3 = 1.12 (s, 3H), 1.20 (s, 3H), 1.67 (s, 3H), 1.93 (s, 3H), 2.16 (s, 3H), 2.37 (s, 3H), .76 (m, 8H), 2.97 (d, J: 6.8 Hz, 1H), 3.36-3.41 (m, 1H), 3.54-3.62 (m, 1H), 3.710, J = 4.4 Hz, 4H), 3.78(s, 4H), 3.84(s, 8H), 4.16- 4.20(m, 3H), 4.41-4.57(m, 3H), 4.70(s, 1H), 4.94(d, J = 9.2 Hz, 1H), 5.66(dd, J = 6.8, 3.6 Hz, 1H), .90(m, 1H), 6.17(q, J = 9.2 Hz, 1H), 6.31(d, J = 4.4 Hz, 1H), 6.95 (s, 1H), 7.00 (s, 1H), 7.03-7.09(m, 4H), 7.19-7.63(m, 34H), 7.80(dd, J = 17.6, 7.2 Hz, 2H), 8.12(m, 2H), 8.47(d, J = 4.8 Hz, 4H). Mass: (EM+2H+)/2: found 831. nd 44; 1H NMR (400 MHz, CDC13): a = 1.05 (t, J = 7.2 Hz, 3H), 1.40 (t, J = 7.6 Hz, 3H), 1.85-1.96 (m, 6H), 3.14—3.54 (m, 2H), 3.52—3.57 (m, 2H), 3.65 (s, 4H), 3.80 (s, 8H), 4057—4. 10 (m, 2H), 5.27 (s, 2H), 5.29 (d, J = 16.0 Hz, 1H), 5.73 (d, J = 16.0 Hz, 1H), 6.88 (s, 2H), 6.99 (d, J = 8.4 Hz, 2H), 7.07-7.12(m, 6H), .66 (m, 18H), 7.93 (d ,J = 2.8 Hz, 1H), 8.07 (d, J = 8.4 Hz, 2H), 8.26(d, J = 9.2 Hz, 1H), 8.48 (d, J = 4.8 Hz, 4H). Mass: (EM+2H+)/2: found 661.4.
Compound 45; 1H NMR (CDC13, 300 MHz): 3 = 1.11 (s, 3H), 1.18 (s, 3H), 1.66 (s, 3H), 1.89 (s, 3H), 2.18 (s, 3H), 2.27 (s, 3H), .57 (m, 7H), 2.68 (t, J = 6.6 Hz, 2H), 2.79 (t, J = 5.7 Hz, 2H), 3.25(q, J: 4.8 Hz, 2H), 3.400, J = 5.1 Hz, 2H), 3.54-3.63 (m, 10H), 3.700, J = 4.5 Hz, 4H), 3.77(s, 4H), 3.81(s, 8H), 4.18(d, J = 8.8 Hz, 1H), 4.28(d, J = 8.8 Hz, 1H), 4.39-4.47(m, 3H), 4.94(d, J = 9.0 Hz, 1H), 5.43(d, J = 3.6 Hz, 1H), 5.66(d, J = 6.6 Hz, 1H), 5.90(dd, J: 8.4, 3.6 HZ, 1H), 6.16(t, J: 9.0 HZ, 1H), 6.28(S, 1H), 7.06-7.09(m, 8H), 7.22-7.61(m, 33H), 7.79(dd, J = 7.5 Hz, 2H), 8.11(dd, J = 7.5 Hz, 2H), 8.46(d, J = 4.5 Hz, 4H). Mass: (EM+2H+)/2: found 978.
Compound 46: 1H NMR (400 MHZ, CDC13): 5 = 1.12 (S, 3H), 1.19 (S, 3H), 1.68 (S, 3H), 1.87-1.88 (m, 1H), 1.90 (s, 3H), 2.07—2.13 (m, 1H), 2.20 (s, 3H), .35 (m, 1H), 2.42 (s, 3H), 2.56 (t, J = 6.8 Hz, 2H), 2.66-2.67 (m, 1H), 2.80 (t, J = 6.8 Hz, 2H), 3.25—3.31 (m, 2H), 3.79-3.96 (m, 18H), 4.11 (S, 2H), 4.20 (d, J: 8.8 HZ, 1H), 4.22-4.28 (m, 2H), 4.31 (d, J = 8 HZ, 1H), 4.40-4.45 (m, 1H), 4.95 (d, J: 8.4 HZ, 1H), 5.48 (d, J: 3.6 HZ, 1H), 5.68 (d, J = 6.8 Hz, 1H), 5.94 (dd, J = 8.4, 3.6 Hz, 1H), 6.22 (t, J = 8.8 Hz, 1H), 6.27 (s, 1H), 6.89 (d, J = 8.4 Hz, 2H), 7.08-7.13 (m, 4H), 7.20 (d, J = 7.6 Hz, 2H), 7.27-7.66 (m, 21H), 7.77 (d, J = 7.2 Hz, 2H), 8.14 (d, J = 7.2 Hz, 2H), 8.41 (d, J = 4.8 Hz, 2H), 8.49 (d, J = 4.8 Hz, 2H).
Mass: (EM+2H+)/2: found 857.8.
Compound 47; 1H NMR (400 MHz, CDC13): a = 1.04 (t, J = 7.2 Hz, 3H), 1.30 (s, 9H), 1.44 (t, J = 7.6 Hz, 3H), 1.86-1.97 (m, 4H), 2.03-2.08 (m, 2H), 3.18-3.24 (m, 2H), 3.36- 3.40 (m, 2H), 3.85(s, 4H), 3.95 (s, 6H), 4.26 (s, 2H), 5.30—5.34 (m, 3H), 5.78 (d, 1.04, J: 3.6 Hz, 3H), 7.09—7.13 (m, 2H), 7.17—7.21 (m, 2H), 7.5 (d, J = 8 Hz, 2H), 7.60-7.76 (m, 9H), 7.84 (d, J = 8.8 Hz, 2H), 8.0 (d, J = 2.4Hz, 1H), 8.28-8.32 (m, 4H), 8.48 (d J = 7.6 Hz, 2H), 8.55 (d ,J = 6.8 Hz, 2H). Mass: (EM+2H+)/2; found 626.8.
Compound 48: 1H NMR(CDC13, 400 MHz): 3 = 0.88(m, 6H), 1.030, J = 6.8 Hz, 3H), 1.39(t, J = 7.2 Hz, 3H), 1.90(m, 2H), 2.23(br, 5H), 2.93(br, 2H), , 3H), , 2H), 3.41-3.78(m, 21H), 3.91(m, 2H), 4.11(s, 2H), 4.33(s, 2H), 4.56(s, 2H), 5.25(s, 2H), 5.32(d, J = 18.4 Hz, 1H), 5.76(d, J = 16.4 Hz, 1H), 6.74(s, 2H), 6.98(s, 2H), .51(m, 30H), 7.69(m, 4H), 7.95(s, 2H), 8.17(d, J = 6 Hz, 2H), 8.30(d, J = 8.8 Hz, 1H).
Compound 49; 1H NMR (300 MHz, CDC13): a = 1.05 (t, J = 7.2 Hz, 3H), 1.25 (m, 3H), 1.41 (t, J = 7.5 Hz, 3H), 1.69 (m, 11H), 1.83-2.02 (m, 4H), 2.27 (m, 2H), 2.84 (m, 1H), 3.00 (d, J = 5.7 Hz, 2H), 3.14—3.21 (m, 3H), 3.64 (s, 3H), 3.76 (s, 4H), 3.84 (s, 8H), 4.15 (s, 1H), 4.57 (s, 1H), .80 (m, 1H), 5.29 (d, J = 4.8 Hz, 2H), 5.32 (d, J = 14.7 Hz, 1H), 5.76 (d, J = 16.5 Hz, 1H), 6.95 (s, 2H), 7.10-7.14(m, 4H), .33 (m, 6H), 7.43-7.46 (m, 4H), 7.57-7.62 (m, 4H), .70 (m, 2H), 7.96 (d, J = 2.1 Hz, 1H), 8.17 (d, J = 8.1 Hz, 2H), 8.29 (d, J = 9.0 Hz, 1H), 8.52 (d, J = 5.1 Hz, 4H). Mass: (EM+2H+)/2; found 730.
Compound 50; 1H NMR (CD013, 400 MHz): 3 = 0.88(m, 2H), 1.13 (s, 3H), 1.21 (s, 3H), 1.22-1.30(m, 4H), 1.68 (s, 3H), 1.69-1.91(m, 6H), 1.91 (s, 3H), 2.05-2.32(m, 2H), 2.21 (s, 3H), 2.42 (s, 3H), 2.55—2.97 (m, 6H), 2.76 (t, J: 6.8 Hz, 2H), 3.14—3.54 (m, 8H), 3.64(s, 4H), 3.67(s, 8H), 3.86 (t, J = 5.6 Hz, 2H), 3.95 (t, J = 6.0 Hz, 2H), 4.10-4.12(m, 2H), 4.09 (d, J = 4.4 Hz, 1H), 4.19 (d, J = 4.4 Hz, 1H), 4.29-4.45(m, 3H), 4.95-4.97(m, 2H), 5.45(d, J = 4.0 Hz, 1H), 5.67(d, J = 7.2 Hz, 1H), 5.92(dd, J = 8.4, 4.0 Hz, 1H), 6.19(t, J = 9.2 Hz, 1H), 6.29(s, 1H), 6.43-6.45 (m, 1H), 6.82 (s, 2H), 6.82-7.14(m, 5H), 7.27-7.63(m, 26H), 7.80(d, J = 7.2 Hz, 2H), 8.13(d, J = 7.2 Hz, 2H), 8.50(d, J = 4.8 Hz, 4H). Mass: (EM+2H+)/2: found 937.
Compound 51: 1H NMR (300 MHz, CDCl3): 8 = 1.05 (t, J = 7.2 Hz, 3H), 1.25-1.40 (m, 12H), 1.83-1.91 (m, 6H), 3.16-3.23 (m, 2H), 3.47-4.05 (m, 28H), , 2H), 4.77(s, 2H), 5.28-5.34 (m, 3H), 5.76 (d, J: 16.2 Hz, 1H), 6.86 (s, 2H), 7.01-7.13 (m, 5H), 7.60-7.66 (m, 16H), 7.96 (d ,J= 2.4 Hz, 1H), 8.08 (d, J: 8.7 Hz, 2H), 8.28(d, J: 9.3 Hz, 1H), 8.48 (d, J = 4.2 Hz, 4H). Mass: (EM+2H+)/2: found 760.4.
Zn-DPA conjugates of each of the compounds described above, denoted as Zn-DPA-(Compound number), were prepared following the ure described below.
More ically, each of Compounds 1-51 was mixed with 2 molar equivalents of zinc nitrate in a solution containing a solvent e of dichloromethane and methanol (1:1) at room temperature and stirred for one hour. Removal of the solvent under vacuum yielded the corresponding Zn-DPA conjugate.
The analytical data of l Zn-DPA conjugates are shown below as representative examples: Zn-DPA-18 [i 1H NMR (700 MHZ, DMSO-dg): 5 = 0.86 (t, J = 7.0 HZ, 3H) , 1.25 (t, J = 7.7 Hz, 3H) = 7.7 Hz, 2H), 1.79-1.89 (m, 4H), 3.16 (q, J = 7.7 Hz, 2H), 3.20 (q, , , J J: 7.7 Hz, 2H), 3.75 (d, J: 16.1 Hz, 4H), 3.84 (s, 4H), 4.11 (br, 2H), 4.33 (d, J: 16.1 Hz, 4H), 5.31 (S, 2H), 5.41 (S, 2H), 6.48 (br, 1H), 6.51 (S, 1H), 6.86 (S, 1H), 6.99 (S, 2H), 7.02 (S, 1H), 7.44 (t, J = 7.7 Hz, 1H), 7.54 (d, J: 7.7 Hz, 4H), 7.60 (t, J = 7.0 Hz, 4H), 7.65 (d, J = 7.7 Hz, 1H), 7.71 (d, J = 7.7 Hz, 1H), 7.65 (dd, J = 9.1, 2.1 Hz, 2H), 8.05 (t, J = 7.7 Hz, 4H), 8.14 (s, 1H), 8.21 (d, J = 9.1 Hz, 1H), 8.37 (s, 1H), 8.63 (d, J = 5.6 Hz, 3H), 8.95 (s, 1H).
Mass: (EM+Zn+2H+)/2: found 595, (EM+2Zn+2H+)/2: found 627. -g 11 g: 1H NMR (700 MHZ, DMSO-dg): 5 = 0.86 (t, J = 7.0 HZ, 3H) , 1.25 (t, J = 7.7 Hz, 3H) = 7.7 Hz, 2H), 3.48 (s, 2H), 3.73 (d, J = 16.1 Hz, , 1.79-1.88 (m, 6H), 3.16 (q, J 4H), 3.81 (br, 4H), 4.10 (br, 1H), 4.32 (d, J: 16.1 HZ, 4H), 4.73 (S, 2H), 5.32 (s, 2H), 5.41 (s, 2H), 6.52 (s, 1H), 6.86 (s, 1H), 6.98 (s, 2H), 7.29 — 7.31 (m, 1H), 7.37 (d, J = 7.7 Hz, 2H), 7.40 (t, J = 7.7 Hz, 4H), 7.47 (t, J = 7.7 Hz, 2H), 7.52 (d, J = 7.7 Hz, 4H), 7.58 — 7.62 (m, 6H), 7.61 (d, J = 7.7 Hz, 2H), 7.76 (t, J: 7.7 Hz, 2H), 7.93 (d, J = 9.1 Hz, 1H), 8.03 (t, J = 7.7 Hz, 4H), 8.20 (d, J = 5.6 Hz, 1H), 8.42 (s, 1H), 8.63 (d, J = 5.6 Hz, 4H) , 8.84 (br, 1H).
Mass: (EM+Zn+2H+)/2: found 678, (EM+2Zn+2H+)/2: found 710.
Zn-DPA-j 17!: 1H NMR (700 MHZ, DMSO-dg): 5 = 0.85 (t, J = 7.0 HZ, 3H) , 1.25 (t, J = 7.7 Hz, 3H) = 7.7 Hz, 3H), 3.53 , 1.74-1.88 (m, 6H), 3.15 (q, J (s, 1H), 3.69 — 3.83 (m, 8H), 3.94 (br, 1H), 4.16 (br, 1H), 4.30 — 4.38 (m, 4H), 4.73 (s, 1H), 4.88 (s, 1H), 5.30 (s, 2H), 5.40 (s, 2H), 6.51 (s, 1H), 6.54 — 6.88 (m, 2H), 6.93 (s, 1H), 7.02 (s, 1H), 7.30 — 7.64 (m, 17H), 7.68 (d, J = 7.7 Hz, 2H), 7.75 (d, J: 9.1 Hz, 1H), 8.05 (t, J: 7.7 Hz, 4H), 8.15 (s, 1H), 8.18 — 8.21 (m, 3H), 8.64 (d, J = 5.6 HZ, 4H). Mass: (EM+Zn+2H+)/2: found 670, (EM+2Zn+2H+)/2: found 702.
Zn-DPA-131 !: 1H NMR (700 MHZ, DMSO-dg): 5 = 0.85 (t, J = 7.0 HZ, 3H) , 1.19 (t, J = 7.0 Hz, 2H) 1.24 — 3.15 (m, 2H), 3.50 (br, , (t, J: 7.7 Hz, 1H) , 1.82-1.84 (m, 6H), 3.07 1H), 3.60 (br, 1H), 3.71 (t, J: 17.5 HZ, 4H), 3.81 (s, 4H), 4.12 — 4.14 (In, 2H), 4.31 (t, J: 14.7 Hz, 4H), 4.65 (s, 1H), 4.82 (s, 1H), 5.29 (s, 2H), 5.41 (s, 2H), 6.51 (s, 1H), 6.86 (s, 1H), 6.99 (s, 2H), 7.26 — 7.72 (m, 20H), 7.72 (d, J = 7.7 Hz, 1H), 8.03 (t, J = 7.7 Hz, 3H), 8.13 (t, J = 7.7 HZ, 1H), 8.63 (d, J = 4.9 HZ, 4H). Mass: (EM+Zn+2H+)/2: found 618, (EM+2Zn+2H+)/2: found 650.
Plasma ity of Zn-DPA Conjugates Zn-DPA conjugates, Zn-DPA-(S), Zn-DPA-(l2), Zn-DPA-(25), Zn-DPA-(26), and Zn-DPA-(42) prepared from the corresponding compounds 8, 12, 25, 26, and 42, respectively, were incubated in mouse plasma at 37 0C for up to 24 hours to assess the stability of these conjugates.
A sample was analyzed with a High Performance Liquid Chromatography system to determine the concentration of a test ate at one of four time points (i.e., 0, 3 hours, 6 hours, and 24 . The percentages of the test conjugate remaining at 3, 6, and 24 hours after incubation in the plasma were determined. The results are shown in Table 1 below. A higher percentage indicates greater ity. Among the five test conjugates, Zn-DPA-(26) and Zn-DPA-(42) prepared from Compounds 26 and 42, respectively, were the most stable.
After 24 hours of incubation, it was found that 95% or higher of these conjugates remained in the plasma.
Table 1. Stability ntage remaining) of Zn-DPA conjugates, Zn-DPA-(S), Zn-DPA-(12), Zn-DPA-(25), Zn-DPA-(26), and Zn-DPA-(42) Zn conjugates time =3 hr time = 6 hr time = 24 hr Zn-DPA-(S) 90 82 - Zn-DPA-(12) 100 84 76 - Zn-DPA-(25) 100 90 85 - -(26) 100 99 97 95 Zn-DPA-(42) 100 99 98 97 Growth Inhibition of Cancer Cells Cell culture SCM-l, MiaPaca2 and Colo205 cells were grown in RPMI 1640 ll Park Memorial Institute) medium (RPMI; Gibco), supplemented with 10% fetal bovine serum (FBS; Gibco). Detroit551 cells were grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% PBS 50 U/mL of penicillin and streptomycin, and 1% Nonessential amino acids (NEAA; Gibco).
Zn-DPA conjugates, Zn-DPA-(S), Zn-DPA-(12), Zn-DPA-(25), Zn-DPA-(26), and Zn-DPA-(42) prepared from the corresponding compounds 8, 12, 25, 26, and 42, tively, were used to inhibit the growth of human cancer cells (SCM-l, Colo205, MiaPaca2) and human embryonic skin fibroblast cells Detroit551 following the ures bed below.
Cell viability assay Cell viability was examined by the MTS assay (Promega, n, WI, USA). More specifically, cells were grown (2500~3000 cells/well) in a flat bottomed 96-well plate for 24 hours. A medium was added along with a test nd at a pre-determined concentration. The cells were further incubated for 72 hours. At the end of the incubation, the medium was removed and diluted with 100 pl of a 3-(4,5-dimethylthiazolyl)(3- carboxymethoxyphenyl)(4-sulfophenyl)-2H-tetrazolium/phenazine methosulfate mixture.
The cells were again incubated for 1.5 hours at 37 0C in a humidified incubator with 5% C02 to allow viable cells to convert the tetrazolium salt into formazan. The conversion was determined by measuring the absorbance at 490 nm using a BioTek PowerWave-X Absorbance Microplate Reader.
The data obtained were normalized using a vehicle (dimethyl Xde, DMSO)- d control (100% viability) and a background control (0% viability) to verify growth inhibition. The IC50 value is defined as the tration of a compound that induces a 50% reduction in cell viability in comparison with a vehicle-treated control. These values were calculated using ad Prism version 4 software (San Diego, CA, USA).
The IC50 values for five Zn-DPA conjugates, i.e., Zn-DPA-(S), -(12), Zn-DPA-(25), Zn-DPA-(26), and Zn-DPA-(42), were shown in Table 2 below. Also shown in this table are IC50 values for two anticancer compounds, i.e., SN-38 and CPT-l 1, for comparison. See below for the structures of these two compounds. Note that Zn-DPA-(S), Zn-DPA-(12), Zn-DPA-(25), Zn-DPA-(26), and Zn-DPA-(42), and CPT-ll are prodrugs of the anti-cancer nd SN-38. Zn-DPA-(26), -(42), and CPT-ll contained SN- 38 at about 24%, 18%, and 58%, respectively.
G “-0 WO 79299 Table 2. IC50 values of Zn-DPA ates, Zn-DPA-(S), Zn-DPA-(12), -(25), Zn-DPA-(26), Zn-DPA-(42), and two anticancer compounds (SN-38 and CPT-l 1) against human cancer cells 05, SCM-l, MiaPaca2) and human embryonic skin fibroblast Detroit 551.
Conjugates or Colo205 SCM-l MiaPaca2 Detroit 551 Compounds Zn-DPA-(S) 0.42 0.64 - 9.3 Zn-DPA-(12) 0.12 0.94 - 2.9 -(25) 0.58 1 - 7.2 Zn-DPA-(26) 1.1 2.6 0.12 >10 Zn-DPA-(42) 3.7 - 0.55 >10 SN-38 0.14 0.64 0.02 4 CPT-11 >10 >10 3.6 >10 >“unit in (HM) In Vivo Antitumor Assay Two Zn-DPA conjugates, Zn-DPA-(26) and Zn-DPA-(42) were ted to an in vivo antitumor assay against Colo205 or MiaPaca2 tumors growing in nude mice following the procedure described below.
More specifically, Colo205 or MiaPaca2 cells were cultured and maintained in a flask with a RPMI-1640 medium, which was supplemented with 10% PBS. The cells were harvested and innoculated (1x106 cells) subcutaneously into the left flank of a adult male nude mouse. Tumor-bearing mice were grouped at the mean tumor volume of approximately 200 mm3. Tumor dimensions were measured with a digital caliper, and the tumor volume in mm3 was ated by the formula: Volume = (length x width A2)/2.
The mice were housed in sterilized cages equipped with an air filter and sterile bedding materials at the Laboratory Animal Center of National Health Research Institutes.
All mice were fed with sterilized water and chow at libitum under 12-hour light/12-hour dark cycle throughout the study.
Several dosages of Zn-DPA-(26) and Zn-DPA-(42), were used in this assay, i.e., 40 mg/kg, 20 mg/kg, and 10 mg/kg, p < 0.05 vs. vehicle control by one-way ANOVA analysis and the Newman-Keuls multiple comparison test. Both Zn-DPA-(26) and Zn-DPA-(42) in the mixture of 10% DMSO/20% Cremophor EL/70% dextrose were intravenously administered in a regimen of once daily for five consecutive days when dosed at 20 mg/kg or mg/kg, or of twice per week, for two weeks when dosed at 40 mg/kg; CPT-ll (40 mg/kg) was intravenously administered twice a week for two weeks; and SN—38 (10 mg/kg) in the mixture of % DMSO/20% Cremophor EL/10% NazCO3/60% dextrose was intravenously administered at once daily for five consecutive days for two weeks.
Although the amounts of SN-38 contained in Zn-DPA-(26) and Zn-DPA-(42) were only 24% and 18%, respectively (as compared to 58% ned in CPT-l 1), these two Zn- DPA conjugates were found to unexpectedly show much greater antitumor activities than those of SN-38 and CPT-ll tested in the C010205 tumor xenograft mouse model. More specifically, it was found that Zn-DPA-(26) ctedly showed much higher antitumor activities at dosages of 10 mg/kg and 40 mg/kg, compared to those of SN-38 at 10 mg/kg and CPT-ll at 40 mg/kg; and Zn-DPA-(42), also unexpectedly, showed antitumor ties in a dose-dependent manner and much higher antitumor activities at all three dosages, compared to those of SN-38 at 10 mg/kg and CPT-ll at 40 mg/kg.
Moreover, in the MiaPaca2 tumor xenograft mouse model, Zn-DPA-(42) unexpectedly showed antitumor activities in a dose-dependent manner and much higher antitumor activities at s of 40 mg/kg, 20 mg/kg, and 10 mg/kg, ed to those of SN-38 at 10 mg/kg and CPT-ll at 40 mg/kg.
OTHER EMBODIMENTS All of the features disclosed in this ication may be combined in any combination. Each feature sed in this specification may be replaced by an alternative feature serving the same, equivalent, or similar purpose. Thus, unless expressly stated otherwise, each feature disclosed is only an example of a generic series of equivalent or similar features.
From the above description, one skilled in the art can easily ain the essential characteristics of the present ion, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions. For example, nds structurally analogous to the compounds of this invention also can be made, screened for their efficacy in ng a condition that relates to cells containing inside-out phosphatidylserine. Thus, other embodiments are also within the claims.

Claims (23)

WHAT IS CLAIMED IS:
1. A compound of formula (I): B1 B2 Z X L1 L2 A5 A6 N N A1 A2 A3 A4 W1 W2 W3 W4 W5 W6 W7 W8 in which each of A1, A2, A3, A4, A5, and A6, independently, is a C1-C6 bivalent aliphatic radical; B1 is a C1-C6 bivalent aliphatic radical, a C1-C6 nt heteroaliphatic radical, a bivalent aryl radical, or a bivalent heteroaryl radical; B2 is a bond, a C1-C6 bivalent aliphatic radical, a C1-C6 bivalent heteroaliphatic radical, a bivalent aryl radical, a bivalent heteroaryl radical, D1-NR1-C(O)-D2, D1-C(O)NR1- D2-NR1’-C(O)-D3, D1-D2-C(O)-NR1-C(O)-D3, or D1-D2-D3, each of D1, D2, D3, independently, being a C1-C6 bivalent aliphatic radical, a C1-C6 bivalent aliphatic radical, a bivalent aryl l, a bivalent heteroaryl radical, a C1-C10 bivalent aralkyl l, or a C1-C10 nt heteroaralkyl radical, and each of R1 and R1’, ndently, being H, a C1-C6 bivalent heteroaliphatic radical, a bivalent aryl radical, a bivalent heteroaryl radical, or a C1-C10 bivalent aralkyl radical; L1 is a bond, NR2, NR2C(O), NR2C(S), NR2CR3R4, NR2SO2, NR2C(O)NR3, or )NR3, each of R2, R3, and R4, independently, being H, a C1-C6 monovalent aliphatic radical, a C1-C6 monovalent heteroaliphatic radical, a monovalent aryl radical, a monovalent heteroaryl radical, a C1-C14 monovalent aralkyl l, a C1-C14 monovalent heteroaralkyl radical, C(s)R’, or , in which R’ is a C1-C6 monovalent aliphatic radical, a C1-C6 monovalent heteroaliphatic radical, a monovalent aryl l, a monovalent heteroaryl l, a C1-C14 monovalent aralkyl radical, or a C1-C14 monovalent heteroaralkyl radical; L2 is NR2, ), NR2C(S), NR2CR3R4, NR2SO2, NR2C(O)NR3, NR2C(S)NR3, or C(O), each of R2, R3, and R4, independently, being H, a C1-C6 monovalent aliphatic radical, a C1-C6 monovalent heteroaliphatic radical, a monovalent aryl radical, a monovalent heteroaryl radical, a C1-C14 monovalent aralkyl radical, a C1-C14 monovalent heteroaralkyl radical, C(s)R’, or , in which R’ is a C1-C6 monovalent aliphatic radical, a C1-C6 monovalent heteroaliphatic radical, a monovalent aryl l, a lent heteroaryl radical, a C1-C14 monovalent aralkyl radical, or a C1-C14 monovalent heteroaralkyl l; each of W1, W2, W3, W4, W5, W6, W7, and W8, independently, is N or CH; X is a bond, O, S, or NR6, R6 being H, a C1-C6 lent aliphatic radical, a C1-C6 monovalent heteroaliphatic radical, a monovalent aryl radical, a monovalent heteroaryl radical, a C1-C14 lent l radical, or a C1-C14 monovalent heteroaralkyl radical; Y is an aryl ring or a heteroaryl ring; and Z is an anticancer therapeutic moiety connected to L2 via a cleavable bond, wherein each of the aliphatic radical, the heteroaliphatic radical, the aralkyl radical, and the heteroaralkyl radical is unsubstituted or substituted with halo, cyano, amino, hydroxyl, nitro, sulfhydryl, C1-C6 alkoxy, C1-C6 alkylamino, C1-C12 dialkylamino, and C1-C6 haloalkyl; and each of the aryl radical and the heteroaryl radical is unsubstituted or substituted with halo, cyano, amino, hydroxyl, nitro, dryl, a C1-C6 tic radical, a C1-C6 heteroaliphatic radical, or a haloaliphatic radical.
2. The compound of claim 1, wherein Y is .
3. The compound of claim 1, wherein each of W1, W2, W3, and W4 is N, and each of W5, W6, W7, and W8 is CH.
4. The compound of claim 1, wherein each of W1, W2, W3, and W8 is N, and each of W4, W5, W6, and W7 is CH.
5. The compound of claim 1, wherein each of A1, A2, A3, A4, A5, and A6 is methylene.
6. The compound of claim 1, wherein B1 is ethylene, propylene, butylene, or hexylene.
7. The compound of claim 1, wherein X is O or NH.
8. The compound of claim 1, wherein L2 is C(O).
9. The compound of claim 1, n L1 is a bond, NH, NHCH2, NHC(O), NHSO2, NHC(O)NH,
10. The compound of claim 1, wherein B2 is a bond, ethylene, phenylene,
11. The nd of claim 1, wherein Z is O HO N N O N N HO O N N N O O O O , N O O O O O , H HO OH , , or .
12. The compound of claim 11, wherein Y is
13. The compound of claim 11, wherein each of A1, A2, A3, A4, A5, and A6 is methylene.
14. The compound of claim 11, wherein X is O or NH.
15. The compound of claim 12, wherein each of A1, A2, A3, A4, A5, and A6 is ene; X is O or NH; and L2 is C(O).
16. The compound of claim 1, wherein the compound is one of the following compounds: 5959 N O N HO Et N N O O N N N N N N N O N HO Et N N N N N N 6161 \N \N \N \N ‘/‘/ l/l/ 6262 6363 N N O O Et O O N N O N N N N HO N N Et 6565 6666 6767 6868 / / / / 6969 7070
17. The compound of claim 16, wherein the nd is one of the following compounds: N O HO Et O N H H N N O O N N N N N N 7373 7474 \N \N \N \N
18. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a complex of a metal ion and a compound of claim 1, wherein the metal ion is a cation having two or more charges.
19. The pharmaceutical composition of claim 18, wherein the metal ion is Zn2+, Cu2+, Ca2+, Mg2+, Mn2+, Ni2+, Co2+, Fe2+, Cd2+, or a combination thereof.
20. Use of an effective amount of a compound of claim 1 in the manufacture of a medicament for treating a condition associated with cells containing inside-out phosphatidylserine, wherein the condition is .
21. The use of claim 20, wherein the compound is a metal x formed of a compound of a (I) and a metal ion.
22. The use of claim 21, wherein the metal ion is Zn2+, Cu2+, Ca2+, Mg2+, Mn2+, Ni2+, Co2+, Fe2+, Cd2+, or a combination thereof.
23. The compound of claim 1, substantially as herein described with reference to any one of the Examples thereof.
NZ726105A 2014-05-22 2015-05-18 Dipicolylamine derivatives and their pharmaceutical uses NZ726105B2 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US201462001725P 2014-05-22 2014-05-22
US62/001,725 2014-05-22
PCT/US2015/031386 WO2015179299A1 (en) 2014-05-22 2015-05-18 Dipicolylamine derivatives and their pharmaceutical uses

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NZ726105A NZ726105A (en) 2020-11-27
NZ726105B2 true NZ726105B2 (en) 2021-03-02

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