Claims
- 1. A pharmaceutical composition comprising a therapeutically effective amount of an organic molecule capable of inhibiting protein-protein interaction between calcineurin and NFAT, and a pharmaceutically acceptable carrier.
- 2. The pharmaceutical composition of claim 1, wherein the agent inhibits dephosphorylation of NFAT by calcineurin.
- 3. The pharmaceutical composition of claim 1, wherein the molecular weight of the organic molecule is less than 2500 Da.
- 4. The pharmaceutical composition of claim 1, wherein the molecular weight of the organic molecule is between about 100 and 2000 Da.
- 5. The pharmaceutical composition of claim 1, wherein the molecular weight of the organic molecule is between about 200 and 1500 Da.
- 6. The pharmaceutical composition of claim 1, wherein the molecular weight of the organic molecule is between about 300 and 1000 Da.
- 7. The pharmaceutical composition of claim 1, wherein the organic molecule binds calcineurin with an affinity constant of at least about 2×104 M−1.
- 8. The pharmaceutical composition of claim 1, wherein the organic molecule binds calcineurin with an affinity constant of at least about 106 M−1.
- 9. The pharmaceutical composition of claim 1, wherein the organic molecule binds calcineurin with an affinity constant of at least about 107 M−1.
- 10. The pharmaceutical composition of claim 1, wherein the organic molecule binds calcineurin with an affinity constant of at least about 108 M−1.
- 11. The pharmaceutical composition of claim 1, wherein the organic molecule is a compound selected from the group consisting of:
formula (I): 10wherein: R1 is hydrogen, C1-C20 alkyl optionally substituted with 1-20 R6, C3-C8 cycloalkyl optionally substituted with 1-3 R6, aryl optionally substituted with 1-4 R6, heterocyclyl optionally substituted with 1-3 R6; heteroaryl optionally substituted with 1-4 R6; C2-C8 alkenyl, or C2-C8 alkynyl, cyano, nitro, carboxy, carbo(C1-C6)alkoxy, trihalomethyl, halogen, C1-C6 alkoxy, hydroxy, aryloxy, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, or ureido; R2 is C1-C20 alkyl optionally substituted with 1-20 R6, C3-C8 cycloalkyl optionally substituted with 1-3 R6, aryl optionally substituted with 1-4 R6, heterocyclyl optionally substituted with 1-3 R6, heteroaryl optionally substituted with 1-4 R6, C1-C6 alkoxy, or hydroxy; R3 is hydrogen or halogen;
R4 is hydrogen, C1-C20 alkyl optionally substituted with 1-20 R6, C3-C8 cycloalkyl optionally substituted with 1-3 R6, aryl optionally substituted with 1-4 R6, heterocyclyl optionally substituted with 1-3 R6, heteroaryl optionally substituted with 1-4 R6, or halogen; R is NR7, O or S; R6 is halogen, hydroxy, oxo, nitro, haloalkyl, alkyl, alkaryl, aryl, aralkyl, alkoxy, aryloxy, amino, alkyl amino, dialkylamino, aryl amino, diarylamino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano, mercapto or ureido; and R7 is C1-C6 alkyl; formula (II): 11wherein: R1 and R2 are each independently hydrogen, halogen, amino, C1-C6alkylamino, di(C1-C6)alkylamino, arylamino, diarylamino, or 4,4-dimethyl-2,6-dioxocyclohexyl; R3 is NR11 or O; R4, R5 and R8 are each independently hydrogen, C1-C6 alkyl, halogen, hydroxy, nitro, haloalkyl, alkaryl, aryl, aralkyl, alkoxy, aryloxy, amino, alkyl amino, dialkylamino, aryl amino, diarylamino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano, mercapto or ureido; R6 is hydrogen, halogen, or when taken together with R7 forms a double bond between the carbon atoms to which they are attached; R7 is hydrogen, halogen, or when taken together with R6 forms a double bond between the carbon atoms to which they are attached; R9 is OR13, or when taken together with R10 forms a double bond between the carbon and nitrogen atoms to which they are attached; R10 is hydrogen, or when taken together with R9 forms a double bond between the carbon and nitrogen atoms to which they are attached; R11 is SO2R12; and R12 is aryl optionally substituted with alkyl; R13 is alkyl or aryl; and formula (III): 12wherein, R1 and R4 are each independently O or NR8; R2 and R3 are each independently hydrogen, halogen, or R2 and R3 together combine to form aryl optionally substituted with 1-4 R9; R5 is hydrogen, halogen, carboxy, acylamino, alkoxycarbonyl, carboxy, alkylcarbonyl, acyloxy, or cyano; R6, R7 and R9 are each independently hydrogen, C1-C6 alkyl, halogen, hydroxy, nitro, haloalkyl, alkaryl, aryl, aralkyl, alkoxy, aryloxy, amino, alkyl amino, dialkylamino, aryl amino, diarylamino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano, mercapto or ureido; R8 is SO2R10; and R10 is aryl optionally substituted with alkyl.
- 12. A method for inhibiting protein-protein interaction between calcineurin and NFAT, comprising:
providing calcineurin and NFAT; providing the pharmaceutical composition of claim 1; and contacting the calcineurin, NFAT, and pharmaceutical composition together, such that the protein-protein interaction between calcineurin and NFAT is inhibited.
- 13. A method of inhibiting an immune response in an animal, comprising administering to the animal the pharmaceutical composition of claim 1.
- 14. A method for treating a disease or condition involving hyperactivity or inappropriate activity of the immune system, comprising:
identifying an animal suffering from a disease or condition involving hyperactivity or inappropriate activity of the immune system; and administering to the animal a therapeutically effective amount of the pharmaceutical composition of claim 1, to thereby treat the disease or condition involving hyperactivity or inappropriate activity of the immune system.
- 15. The method of claim 14, wherein the organic molecule is a compound selected from the group consisting of:
formula (I): 13wherein: R1 is hydrogen, C1-C20 alkyl optionally substituted with 1-20 R6, C3-C8 cycloalkyl optionally substituted with 1-3 R6, aryl optionally substituted with 1-4 R6, heterocyclyl optionally substituted with 1-3 R6; heteroaryl optionally substituted with 1-4 R6; C2-C8 alkenyl, or C2-C8 alkynyl, cyano, nitro, carboxy, carbo(C1-C6)alkoxy, trihalomethyl, halogen, C1-C6 alkoxy, hydroxy, aryloxy, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, or ureido; R2 is C1-C20 alkyl optionally substituted with 1-20 R6, C3-C8 cycloalkyl optionally substituted with 1-3 R6, aryl optionally substituted with 1-4 R6, heterocyclyl optionally substituted with 1-3 R6, heteroaryl optionally substituted with 1-4 R6, C1-C6 alkoxy, hydroxy; R3 is hydrogen or halogen; R4 is hydrogen, C1-C20 alkyl optionally substituted with 1-20 R6, C3-C8 cycloalkyl optionally substituted with 1-3 R6, aryl optionally substituted with 1-4 R6, heterocyclyl optionally substituted with 1-3 R6, heteroaryl optionally substituted with 1-4 R6, or halogen; R5 is NR7, O or S; R6 is halogen, hydroxy, oxo, nitro, haloalkyl, alkyl, alkaryl, aryl, aralkyl, alkoxy, aryloxy, amino, alkyl amino, dialkylamino, aryl amino, diarylamino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano, mercapto or ureido; and R7 is C1-C6 alkyl; formula (II): 14wherein: R1 and R2 are each independently hydrogen, halogen, amino, C1-C6alkylamino, di(C1-C6)alkylamino, arylamino, diarylamino, or 4,4-dimethyl-2,6-dioxocyclohexyl; R3 is NR11 or O; R4, R5 and R8 are each independently hydrogen, C1-C6 alkyl, halogen, hydroxy, nitro, haloalkyl, alkaryl, aryl, aralkyl, alkoxy, aryloxy, amino, alkyl amino, dialkylamino, aryl amino, diarylamino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano, mercapto or ureido; R6 is hydrogen, halogen, or when taken together with R7 forms a double bond between the carbon atoms to which they are attached; R7 is hydrogen, halogen, or when taken together with R6 forms a double bond between the carbon atoms to which they are attached; R9 is OR13, or when taken together with R10 forms a double bond between the carbon and nitrogen atoms to which they are attached; R10 is hydrogen, or when taken together with R9 forms a double bond between the carbon and nitrogen atoms to which they are attached; R11 is SO2R12; and R12 is aryl optionally substituted with alkyl; R13 is alkyl or aryl; and formula (III): 15wherein, R1 and R4 are each independently O or NR8; R2 and R3 are each independently hydrogen, halogen, or R2 and R3 together combine to form aryl optionally substituted with 1-4 R9; R5 is hydrogen, halogen, carboxy, acylamino, alkoxycarbonyl, carboxy, alkylcarbonyl, acyloxy, or cyano; R6, R7 and R9 are each independently hydrogen, C1-C6 alkyl, halogen, hydroxy, nitro, haloalkyl, alkaryl, aryl, aralkyl, alkoxy, aryloxy, amino, alkyl amino, dialkylamino, aryl amino, diarylamino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano, mercapto or ureido; R8 is SO2R10; and R10 is aryl optionally substituted with alkyl.
- 16. The method of claim 14 wherein the disease or condition involving hyperactivity or inappropriate activity of the immune system is selected from the group consisting of: an acute immune disease, a chronic immune disease, and an autoimmune disease.
- 17. A method for treating a disease involving excessive or inappropriate activation of NFAT, or a molecular target thereof, comprising:
identifying an animal suffering from a disease involving excessive or inappropriate activation of NFAT or a molecular target thereof, and administering to the animal a therapeutically effective amount of the pharmaceutical composition of claim 1, to thereby treat the disease involving excessive or inappropriate activation of NFAT or molecular target thereof.
- 18. A process of making an agent that inhibits protein-protein interaction between calcineurin and NFAT, the process comprising:
carrying out a method to identify an agent that inhibits protein-protein interaction between calcineurin and NFAT, wherein the method comprises:
providing a first compound selected from the group consisting of calcineurin or a biologically active derivative thereof, and NFAT or a biologically active derivative thereof; providing a second compound selected from the group consisting of calcineurin or a biologically active derivative thereof, and NFAT or a biologically active derivative thereof, wherein the second compound is different from the first compound, and wherein the second compound is labeled; providing a candidate agent; contacting the first compound, the second compound, and the candidate agent with each other; and determining the amount of label bound to the first compound, wherein a reduction in interaction between the first compound and the second compound as assessed by label bound is indicative of usefulness of the candidate agent in inhibiting protein-protein interaction between calcineurin and NFAT; and manufacturing the agent, to thereby make an agent that inhibits protein-protein interaction between calcineurin and NFAT.
- 19. The process of claim 18, wherein the first compound is calcineurin and the second compound is a biologically active derivative of NFAT.
- 20. The process of claim 18, wherein the biologically active derivative of NFAT comprises the amino acid sequence GPHPVIVITGPHEE.
- 21. A method of manufacturing an agent that inhibits protein-protein interaction between calcineurin and NFAT, the method comprising:
providing an organic compound capable of inhibiting protein-protein interaction between calcineurin and NFAT; providing at least one pharmaceutically acceptable carrier; and combining the organic compound with the pharmaceutically acceptable carrier, to thereby manufacture an agent that inhibits protein-protein interaction between calcineurin and NFAT.
- 22. The method of claim 21, further comprising the step of manufacturing the agent into a form suitable for administration to an animal via a route selected from a group consisting of: oral, parenteral, topical, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, intrasternal.
- 23. A method for inhibiting protein-protein interaction between calcineurin and NFAT, comprising:
providing calcineurin and NFAT; providing an organic molecule capable of inhibiting protein-protein interaction between calcineurin and NFAT, wherein the organic molecule is a compound selected from the group consisting of: formula (I): 16wherein: R1 is hydrogen, C1-C20 alkyl optionally substituted with 1-20 R6, C3-C8 cycloalkyl optionally substituted with 1-3 R6, aryl optionally substituted with 1-4 R6, heterocyclyl optionally substituted with 1-3 R6; heteroaryl optionally substituted with 1-4 R6; C2-C8 alkenyl, or C2-C8 alkynyl, cyano, nitro, carboxy, carbo(C1-C6)alkoxy, trihalomethyl, halogen, C1-C6 alkoxy, hydroxy, aryloxy, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, or ureido; R2 is C1-C20 alkyl optionally substituted with 1-20 R6, C3-C8 cycloalkyl optionally substituted with 1-3 R6, aryl optionally substituted with 1-4 R6, heterocyclyl optionally substituted with 1-3 R6, heteroaryl optionally substituted with 1-4 R6, C1-C6 alkoxy, hydroxy; R3 is hydrogen or halogen; R4 is hydrogen, C1-C20 alkyl optionally substituted with 1-20 R6, C3-C8 cycloalkyl optionally substituted with 1-3 R6, aryl optionally substituted with 1-4 R6, heterocyclyl optionally substituted with 1-3 R6, heteroaryl optionally substituted with 1-4 R6, or halogen; R5 is NR7, O or S; R6 is halogen, hydroxy, oxo, nitro, haloalkyl, alkyl, alkaryl, aryl, aralkyl, alkoxy, aryloxy, amino, alkyl amino, dialkylamino, aryl amino, diarylamino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano, mercapto or ureido; and R7 is C1-C6 alkyl; formula (II): 17wherein: R1 and R2 are each independently hydrogen, halogen, amino, C1-C6alkylamino, di(C1-C6)alkylamino, arylamino, diarylamino, or 4,4-dimethyl-2,6-dioxocyclohexyl; R3 is NR11 or O; R4, R5 and R8 are each independently hydrogen, C1-C6 alkyl, halogen, hydroxy, nitro, haloalkyl, alkaryl, aryl, aralkyl, alkoxy, aryloxy, amino, alkyl amino, dialkylamino, aryl amino, diarylamino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano, mercapto or ureido; R6 is hydrogen, halogen, or when taken together with R7 forms a double bond between the carbon atoms to which they are attached; R7 is hydrogen, halogen, or when taken together with R6 forms a double bond between the carbon atoms to which they are attached; R9 is OR13, or when taken together with R10 forms a double bond between the carbon and nitrogen atoms to which they are attached; R10 is hydrogen, or when taken together with R9 forms a double bond between the carbon and nitrogen atoms to which they are attached; R11 is SO2R12; and R12 is aryl optionally substituted with alkyl; R13 is alkyl or aryl; and formula (III): 18wherein, R1 and R4 are each independently O or NR8; R2 and R3 are each independently hydrogen, halogen, or R2 and R3 together combine to form aryl optionally substituted with 1-4 R9; R5 is hydrogen, halogen, carboxy, acylamino, alkoxycarbonyl, carboxy, alkylcarbonyl, acyloxy, or cyano; R6, R7 and R9 are each independently hydrogen, C1-C6 alkyl, halogen, hydroxy, nitro, haloalkyl, alkaryl, aryl, aralkyl, alkoxy, aryloxy, amino, alkyl amino, dialkylamino, aryl amino, diarylamino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano, mercapto or ureido; R8 is SO2R10; and R10 is aryl optionally substituted with alkyl; and contacting the calcineurin, NFAT, and organic molecule together such that protein-protein interaction between the calcineurin and the NFAT is inhibited.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation-in-part of U.S. patent application Ser. No. 10/066,151, filed on Jan. 31, 2002, which is a continuation of U.S. patent application Ser. No. 09/248,620, filed Feb. 11, 1999, which claims the benefit of U.S. Provisional Application No. 60/074,467, filed Feb. 12, 1998, all of which are incorporated herein by reference in their entirety.
Government Interests
[0002] This invention was made with Government support under National Institutes of Health Grant Nos. AI 40127, AI 43726, GM 38608, and GM 47467. The Government has certain rights in this invention.
Provisional Applications (1)
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Number |
Date |
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60074467 |
Feb 1998 |
US |
Continuations (1)
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Number |
Date |
Country |
Parent |
09248620 |
Feb 1999 |
US |
Child |
10066151 |
Jan 2002 |
US |
Continuation in Parts (1)
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Number |
Date |
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Parent |
10066151 |
Jan 2002 |
US |
Child |
10358052 |
Feb 2003 |
US |