Specific inhibitors of NFAT activation by calcineurin and their use in treating immune-related diseases

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.

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.

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):
• 16
• wherein:
• 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):
• 17
• wherein:
• 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):
• 18
• wherein,
• 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.