Methods and compositions utilizing quinazolinones

• US 6,562,831 B1
• Filed: 11/28/2000
• Issued: 05/13/2003
• Est. Priority Date: 10/27/1999
• Status: Expired due to Fees

• Current Assignee: Cytokinetics, Inc.
• Original Assignee: Cytokinetics, Inc.
• Inventors: Finer, Jeffrey T., Feng, Bainian, Chabala, John C., Smith, Whitney W., Bergnes, Gustave
• Primary Examiner(s): Raymond, Richard L.
• Assistant Examiner(s): TRUONG, TAMTHOM NGO
• Application Number:
  US09/724,644
• Time in Patent Office: 896 Days
• Field of Search: 514/266.3, 514/266.31, 544/284, 544/287, 436/504
• US Class Current: 514/266.3
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07D 239/90 with acyclic radicals attached in position 2 or 3
• CPC Class Codes: C07D 239/91 with aryl or aralkyl radicals attached in position 2 or 3

Abstract

Quinazolinones of formulae 1a, 1b, 1c and 1d are disclosed. They are useful for treating cellular proliferative diseases and disorders associated with KSP kinesin activity.

Claims

1. A method of screening for compounds that bind to KSP kinesin comprising the steps of:

(a) combining a KSP kinesin with a labeled compound, wherein said compound is chosen from the group consisting of;

 

wherein;

R₁ is chosen from hydrogen, alkyl, aryl, alkylaryl, heteroaryl, alkylheteroaryl, substituted alkyl, substituted aryl, substituted alkylaryl, substituted heteroaryl, and substituted alkylheteroaryl;

R₂ and R₂′

are independently chosen from hydrogen, alkyl, oxaalkyl, aryl, alkylaryl, heteroaryl, alkylheteroaryl, substituted alkyl, substituted aryl, substituted alkylaryl, substituted heteroaryl, and substituted alkylheteroaryl;

or R₂ and R₂′

taken together form a 3- to 7-membered ring;

R₃ is chosen from hydrogen, alkyl, aryl, alkylaryl, heteroaryl, alkylheteroaryl, substituted alkyl, substituted aryl, substituted alkylaryl, substituted heteroaryl, substituted alkylheteroaryl, oxaalkyl, oxaalkylaryl, substituted oxaalkylaryl, R₁₅O— and

R₁₅—

NH—

;

R_3′

is chosen from hydrogen, alkyl, aryl, alkylaryl, heteroaryl, alkylheteroaryl, substituted alkyl, substituted aryl, substituted alkylaryl, substituted heteroaryl, substituted alkylheteroaryl and R₁₅—

NH—

;

R_3″

is chosen from alkyl, aryl, alkylaryl, heteroaryl, alkylheteroaryl, substituted alkyl, substituted aryl, substituted alkylaryl, substituted heteroaryl, and substituted alkylheteroaryl;

R₄ is chosen from hydrogen, alkyl, aryl, alkylaryl, heteroaryl, alkylheteroaryl, substituted alkyl, substituted aryl, substituted alkylaryl, substituted heteroaryl, substituted alkylheteroaryl, and R₁₆-alkylene-;

R₅, R₆, R₇ and R₈ are independently chosen from hydrogen, alkyl, alkoxy, halogen, fluoroalkyl, nitro, dialkylamino, alkylsulfonyl, alkylsulfonamido, sulfonamidoalkyl, sulfonamidoaryl, alkylthio, carboxyalkyl, carboxamido, aminocarbonyl, aryl and heteroaryl;

R₁₅ is chosen from hydrogen, alkyl, aryl, alkylaryl, heteroaryl, alkylheteroaryl, substituted alkyl, substituted aryl, substituted alkylaryl, substituted heteroaryl, and substituted alkylheteroaryl;

R₁₆ is chosen from alkoxy, amino, alkylamino, dialkylamino, N-heterocyclyl and substituted N-heterocyclyl;

under conditions suitable for said compound to bind to said KSP kinesin;

(b) adding a candidate agent under conditions suitable for said agent to bind to said KSP kinesin, thereby displacing at least a portion of said compound that had been bound to said KSP kinesin; and

(c) determining the binding of said agent to said KSP by monitoring the amount of said compound either bound to said KSP or displaced therefrom.

2. The method of claim 1, wherein said label directly or indirectly provides a detectable signal.

3. The method of claim 1, wherein said label is selected from the group consisting of a radioisotope, a fluorescent group, an enzyme, a magnetic particle group, a specific binding molecule and a chemiluminescent group.

4. The method of claim 1, wherein, in step (a), said conditions suitable for said compound to bind to said KSP comprise incubating said compound with said kinesin.

5. The method of claim 4, wherein said incubation is performed at a temperature of between 4 and 40°

C.

6. The method of claim 4, wherein after incubating said compound with said kinesin, said method further comprises washing to remove excess compound.

7. The method of claim 1, wherein, in step (b), said conditions suitable for said agent to bind to said KSP kinesin comprise incubating said agent with said kinesin.

8. The method of claim 1, wherein the screening occurs in the presence of microtubules.

18. A method according to claims 1 or 9, wherein said compound has a formula:

19. A method according to claim 18 wherein R₁ is chosen from hydrogen, lower alkyl, substituted lower alkyl, benzyl, substituted benzyl, phenyl, naphthyl and substituted phenyl.

20. A method according to claim 19 wherein R₁ is chosen from hydrogen, ethyl, propyl, methoxyethyl, naphthyl, phenyl, bromophenyl, chlorophenyl, methoxyphenyl, ethoxyphenyl, tolyl, dimethylphenyl, chorofluorophenyl, methylchlorophenyl, ethylphenyl, phenethyl, benzyl, chlorobenzyl, methylbenzyl, methoxybenzyl, tetrahydrofuranylmethyl and (ethoxycarbonyl)ethyl.

21. A method according to claim 18 wherein R₂ is chosen from hydrogen, lower alkyl and substituted lower alkyl, and R₂′

is hydrogen.

22. A method according to claim 21 wherein R₂ is chosen from hydrogen, methyl, ethyl, propyl, methylthioethyl, aminobutyl, (CBZ)aminobutyl, cyclohexylmethyl, benzyloxymethyl, methylsulfinylethyl, methylsulfinylmethyl, hydroxymethyl, benzyl and indolylmethyl.

23. A method according to claim 18, wherein R₃ is chosen from C₁-C₁₃ alkyl;

substituted lower alkyl;

phenyl;

naphthyl;

phenyl substituted with one or more halo, lower alkyl, loweralkoxy, nitro, carboxy, methylenedioxy or trifluoromethyl;

biphenylyl;

benzyl;

phenoxymethyl;

halophenoxymethyl;

phenylvinyl;

heteroaryl;

heteroaryl substituted with lower alkyl; and

benzyloxymethyl.

24. A method according to claim 23 wherein R₃ is chosen from ethyl, propyl, chloropropyl, butoxy, heptyl, butyl, octyl, tridecanyl, (ethoxycarbonyl)ethyl, dimethylaminoethyl, dimethylaminomethyl, phenyl, naphthyl, halophenyl, dihalophenyl, cyanophenyl, halo(trifluoromethyl)phenyl, chlorophenoxymethyl, methoxyphenyl, carboxyphenyl, ethylphenyl, tolyl, biphenylyl, methylenedioxyphenyl, methylsulfonylphenyl, methoxychlorophenyl, chloronaphthyl, methylhalophenyl, trifluoromethylphenyl, butylphenyl, pentylphenyl, methylnitrophenyl, phenoxymethyl, dimethoxyphenyl, phenylvinyl, nitrochlorophenyl, nitrophenyl, dinitrophenyl, bis(trifluoromethyl)phenyl, benzyloxymethyl, benzyl, furanyl, benzofuranyl, pyridinyl, indolyl, methylpyridinyl, quinolinyl, picolinyl, pyrazolyl, and imidazolyl.

25. A method according to claim 18 wherein R₃ is R₁₅—

NH— and

R₁₅ is chosen from lower alkyl;

cyclohexyl;

phenyl; and

phenyl substituted with halo, lower alkyl, loweralkoxy, or lower alkylthio.

26. A method according to claim 25 wherein R₁₅ is chosen from isopropyl, butyl, cyclohexyl, phenyl, bromophenyl, dichlorophenyl, methoxyphenyl, ethylphenyl, tolyl, trifluoromethylphenyl and methylthiophenyl.

27. A method according to claim 18 wherein R₄ is chosen from lower alkyl, substituted lower alkyl, cyclohexyl;

phenyl substituted with hydroxy, lower alkoxy or lower alkyl;

benzyl;

heteroarylmethyl;

heteroarylethyl;

heteroarylpropyl and R₁₆-alkylene-, wherein R₁₆ is amino, lower alkylamino, di(lower alkyl)amino, lower alkoxy, or N-heterocyclyl.

28. A method according to claim 27 wherein R₄ is chosen from methyl, ethyl, propyl, butyl, cyclohexyl, carboxyethyl, carboxymethyl, methoxyethyl, hydroxyethyl, hydroxypropyl, dimethylaminoethyl, dimethylaminopropyl, diethylaminoethyl, diethylaminopropyl, aminopropyl, methylaminopropyl, 2,2-dimethyl-3-(dimethylamino)propyl, 1-cyclohexyl-4-(diethylamino)butyl, aminoethyl, aminobutyl, aminopentyl, aminohexyl, aminoethoxyethyl, isopropylaminopropyl, diisopropylaminoethyl, 1-methyl-4-(diethylamino)butyl, (t-Boc)aminopropyl, hydroxyphenyl, benzyl, methoxyphenyl, methylmethoxyphenyl, dimethylphenyl, tolyl, ethylphenyl, (oxopyrrolidinyl)propyl, (methoxycarbonyl)ethyl, benzylpiperidinyl, pyridinylethyl, pyridinylmethyl, morpholinylethyl, morpholinylpropyl, piperidinyl, azetidinylmethyl, azetidinylpropyl, pyrrolidinylethyl, pyrrolidinylpropyl, piperidinylmethyl, piperidinylethyl, imidazolylpropyl, imidazolylethyl, (ethylpyrrolidinyl)methyl, (methylpyrrolidinyl)ethyl, (methylpiperidinyl)propyl, (methylpiperazinyl)propyl, furanylmethyl and indolylethyl.

29. A method according to claim 18 wherein

R₁ is chosen from lower alkyl, benzyl, substituted benzyl and substituted phenyl;

R₂ is chosen from hydrogen, alkyl, substituted lower alkyl and benzyl;

R₂′

is hydrogen;

R₃ is chosen from substituted phenyl and naphthyl;

R₄ is chosen from substituted alkyl and R₁₆-alkylene-;

R₅ is hydrogen or halo

R₆ is hydrogen, methyl or halo;

R₇ is hydrogen, halo, methyl or trifluoromethyl;

R₈ is hydrogen or halo;

R₁₆ is chosen from di(lower alkylamino), (lower alkyl)amino, amino, N-heterocyclyl and substituted N-heterocyclyl.

30. A method according to claim 1 or 9 wherein

R₁ is benzyl or halobenzyl;

R₂ is chosen from ethyl and propyl;

R₂′

is hydrogen;

R₃ is substituted phenyl;

R_3′

is substituted phenyl;

R_3″

is substituted phenyl,

R₄ is (CH₂)_mOH or (CH₂)_pR₁₆ wherein m is 2 or 3 and p is 1-3;

R₅ is hydrogen;

R₆ is hydrogen;

R₇ is halo;

R₈ is hydrogen;

R₁₆ is chosen from amino, propylamino, and azetidinyl.

31. A method according to claim 30 wherein the stereogenic center to which R₂ and R_2′

are attached is of the R configuration.

32. A method according to claim 1 or 9, wherein said compound has a formula:

33. A method according to claim 32 wherein:

R₁ is chosen from hydrogen, lower alkyl, substituted lower alkyl, benzyl, substituted benzyl, phenyl, naphthyl and substituted phenyl;

R₂ is chosen from hydrogen, lower alkyl and substituted lower alkyl and R₂′

is hydrogen;

R_3′

is chosen from C₁-C₁₃ alkyl;

phenyl;

naphthyl;

phenyl substituted with halo, lower alkyl, lower alkoxy, nitro, methylenedioxy, or trifluoromethyl;

biphenylyl, benzyl and heteroaryl; and

R₄ is chosen from lower alkyl, substituted lower alkyl, cyclohexyl;

phenyl substituted with hydroxy, lower alkoxy or lower alkyl;

benzyl;

heteroarylmethyl;

heteroarylethyl;

heteroarylpropyl and R₁₆-alkylene, wherein

R₁₆ is amino, (lower alkyl)amino, di(lower alkyl)amino, lower alkoxy, or N-heterocyclyl.

34. A method according to claim 1 or 9 wherein

R₁ is chosen from lower alkyl, benzyl, substituted benzyl and substituted phenyl;

R₂ is hydrogen or lower alkyl;

R₂′

is hydrogen;

R₃ is chosen from substituted phenyl and naphthyl;

R₄ is R₁₆-alkylene-hydroxy lower alkyl or carboxy lower alkyl;

R₆ and R₇ are chosen from hydrogen and halo;

R₅ and R₈ are hydrogen;

R₁₆ is chosen from di(lower alkylamino), (lower alkyl)amino, amino, piperidinyl, azetidinyl pyrrolidinyl and morpholinyl.

35. A method according to claim 1 or 9, wherein said compound has a formula:

36. A method according to claim 35 wherein:

R₁ is chosen from hydrogen, lower alkyl, substituted lower alkyl, benzyl, substituted benzyl, phenyl, naphthyl and substituted phenyl;

R₂ is chosen from hydrogen, lower alkyl and substituted lower alkyl and R₂′

is hydrogen; and

R₄ is chosen from lower alkyl, cyclohexyl;

phenyl substituted with hydroxy, lower alkoxy or lower alkyl;

benzyl;

heteroarylmethyl;

heteroarylethyl;

heteroarylpropyl and R₁₆-alkylene, wherein R₁₆ is di(lower alkyl)amino, alkylamino, amino, lower alkoxy, or N-heterocyclyl.

37. A method according to claim 35 wherein

R₁ is chosen from lower alkyl, benzyl, substituted benzyl and substituted phenyl;

R₂ is hydrogen or lower alkyl;

R₂′

is hydrogen;

R₄ is R₁₆-alkylene-;

R₆ and R₇ are chosen from hydrogen and halo;

R₅ and R₈ are hydrogen;

R₁₆ is chosen from di(lower alkylamino), (lower alkyl)amino, amino, pyrrolidinyl, piperidinyl, imidazolyl and morpholinyl.

38. A method according to claims 1 or 9, wherein said compound has a formula:

39. A method according to claim 38 wherein:

R₁ is chosen from hydrogen, lower alkyl, substituted lower alkyl, benzyl, substituted benzyl, phenyl, naphthyl and substituted phenyl;

R₂ is chosen from hydrogen, lower alkyl and substituted lower alkyl and R₂′

is hydrogen;

R_3″

is chosen from C₁-C₁₃ alkyl;

substituted lower alkyl;

phenyl;

naphthyl;

phenyl substituted with halo, lower alkyl, lower alkoxy, nitro, methylenedioxy, or trifluoromethyl;

biphenylyl;

benzyl and heterocyclyl; and

R₄ is chosen from lower alkyl, substituted lower alkyl;

cyclohexyl;

phenyl substituted with hydroxy, lower alkoxy or lower alkyl;

benzyl;

substituted benzyl;

heterocyclyl;

heteroarylmethyl;

heteroarylmethyl;

heteroarylpropyl and R₁₆-alkylene, wherein

R₁₆ is di(lower alkyl)amino, (lower alkyl)amino, amino, lower alkoxy, or N-heterocyclyl.

40. A method according to claim 39 wherein

R₁ is chosen from lower alkyl, benzyl, substituted benzyl and substituted phenyl;

R₂ is hydrogen or lower alkyl;

R₂′

is hydrogen;

R_3″

is chosen from substituted phenyl, heterocyclyl and naphthyl;

R₄ is chosen from substituted benzyl, heterocyclyl substituted lower alkyl and R₁₆-alkylene-;

R₆ and R₇ are chosen from hydrogen and halo;

R₅ and R₈ are hydrogen;

R₁₆ is chosen from di(lower alkylamino), (lower alkyl)amino, amino, pyrrolidinyl, azetidinyl piperidinyl, imidazolyl and morpholinyl.

41. A method according to claim 40 wherein

R₁ is benzyl;

R₂ is ethyl;

R₂′

is hydrogen;

R_3″

is chosen from halophenyl, polyhalophenyl, tolyl, dimethylphenyl, methoxyphenyl, dimethoxyphenyl, cyanophenyl, trifluoromethylphenyl, trifluoromethoxyphenyl, bis(trifluoromethyl)phenyl, carboxyphenyl, t-butylphenyl, methoxycarbonylphenyl, piperidinyl and naphthyl;

R₄ is chosen from substituted benzyl, piperidinyl, hydroxy (lower alkyl) and R₁₆-alkylene-;

R₆ and R₇ are chosen from hydrogen and halo;

R₅ and R₈ are hydrogen;

R₁₆ is chosen from dimethylamino, amino, pyrrolidinyl and piperidinyl.

9. A method of screening for compounds that bind to KSP kinesin comprising the steps of:

(a) combining a KSP kinesin with a labeled candidate agent under conditions suitable for said agent to bind to said KSP kinesin; and

(b) adding a compound under conditions suitable for said compound to bind to said KSP kinesin and displace said agent therefrom, wherein said compound is chosen from the group consisting of;

 

wherein;

R₁ is chosen from hydrogen, alkyl, aryl, alkylaryl, heteroaryl, alkylheteroaryl, substituted alkyl, substituted aryl, substituted alkylaryl, substituted heteroaryl, and substituted alkylheteroaryl;

R₂ and R₂′

are independently chosen from hydrogen, alkyl, oxaalkyl, aryl, alkylaryl, heteroaryl, alkylheteroaryl, substituted alkyl, substituted aryl, substituted alkylaryl, substituted heteroaryl, and substituted alkylheteroaryl;

or R₂ and R₂′

taken together form a 3- to 7-membered ring;

R₃ is chosen from hydrogen, alkyl, aryl, alkylaryl, heteroaryl, alkylheteroaryl, substituted alkyl, substituted aryl, substituted alkylaryl, substituted heteroaryl, substituted alkylheteroaryl, oxaalkyl, oxaalkylaryl, substituted oxaalkylaryl, R₁₅O— and

R₁₅—

NH—

;

R_3′

is chosen from hydrogen, alkyl, aryl, alkylaryl, heteroaryl, alkylheteroaryl, substituted alkyl, substituted aryl, substituted alkylaryl, substituted heteroaryl, substituted alkylheteroaryl and R₁₅—

NH—

;

R_3″

is chosen from alkyl, aryl, alkylaryl, heteroaryl, alkylheteroaryl, substituted alkyl, substituted aryl, substituted alkylaryl, substituted heteroaryl, and substituted alkylheteroaryl;

R₄ is chosen from hydrogen, alkyl, aryl, alkylaryl, heteroaryl, alkylheteroaryl, substituted alkyl, substituted aryl, substituted alkylaryl, substituted heteroaryl, substituted alkylheteroaryl, and R₁₆-alkylene-;

R₅, R₆, R₇ and R₈ are independently chosen from hydrogen, alkyl, alkoxy, halogen, fluoroalkyl, nitro, dialkylamino, alkylsulfonyl, alkylsulfonamido, sulfonamidoalkyl, sulfonamidoaryl, alkylthio, carboxyalkyl, carboxamido, aminocarbonyl, aryl and heteroaryl;

R₁₅ is chosen from hydrogen, alkyl, aryl, alkylaryl, heteroaryl, alkylheteroaryl, substituted alkyl, substituted aryl, substituted alkylaryl, substituted heteroaryl, and substituted alkylheteroaryl;

R₁₆ is chosen from alkoxy, amino, alkylamino, dialkylamino, N-heterocyclyl and substituted N-heterocyclyl; and

(c) determining the binding of said agent to said KSP kinesin by monitoring the amount of labeled compound either bound to KSP or displaced therefrom.

10. The method of claim 9, wherein said label directly or indirectly provides a detectable signal.

11. The method of claim 10, wherein said label is selected from the group consisting of a radioisotope, a fluorescent group, an enzyme, a magnetic particle group, a specific binding molecule and a chemiluminescent group.

12. The method of claim 9, wherein, in step (a), said conditions suitable for said agent to bind to said KSP comprise incubating said agent with said kinesin.

13. The method of claim 12, wherein said incubation is performed at a temperature of between 4 and 40°

C.

14. The method of claim 12, wherein after incubating said agent with said kinesin, said method further comprises washing to remove excess agent.

15. The method of claim 9, wherein, in step (b), said conditions suitable for said compound to bind to said KSP comprise incubating said compound with said kinesin.

16. The method of claim 15 wherein said incubation is performed at a temperature of between 4 and 40°

C.

17. The method of claim 9, wherein the screening occurs in the presence of microtubules.