Regulation of biological events using novel compounds

A method for multimerizing chimeric proteins in cells which comprises:

• (a) providing cells which contain:
• (i) a first recombinant nucleic acid encoding a first chimeric protein which binds to rapamycin or an analog thereof and which comprises at least one FKBP domain and at least one protein domain heterologous thereto, wherein the FKBP domain comprises a peptide sequence selected from:
• (1) a naturally occurring FKBP
• (2) a variant of a naturally occurring FKBP in which up to 10 amino acid residues have been deleted, inserted, or replaced with substitute amino acids,
• (3) an FKBP encoded by a DNA sequence which selectively hybridizes to a DNA sequence encoding an FKBP of (1) or (2);
• (ii) a second recombinant nucleic acid encoding a second chimeric protein which forms a complex with both (a) rapamycin or a rapamycin analog and (b) the first chimeric protein, and which comprises at least one FRB domain and at least one domain heterologous thereto, wherein the FRB domain comprises a peptide sequence selected from:
• (4) a naturally occurring FRB domain,
• (5) a variant of a naturally occurring FRB domain in which up to 10 amino acid residues have been deleted, inserted, or replaced with substitute amino acids,
• (6) an FRB domain encoded by a DNA sequence which selectively hybridizes to a DNA sequence encoding an FRB of (4) or (5);
• and
• (b) contacting the cells with a rapalog which forms a complex containing itself and at least one molecule of each of the first and second chimeric proteins,
• where the rapalog is of the formula:
• wherein
• one of RC7a and RC7b is H and the other is —H, halo, —R2, —OR1, —SR1, —OC(O)R1, —OC(O)NHR1, —NHR1, —NHR1R2, —NHC(O)R1, or —NH—SO2—R1, where R2=aliphatic, heteroaliphatic, aryl, heteroaryl or alkylaryl,
• RC30 is halo, —OR3 or (═O),
• RC24 is ═O, ═NR4 ═NOR4 or ═NNHR4, —NHOR4 or —NHNHR4, —OR4, —OC(O)R4, —OC(O)NR4, halo or —H,
• RC14 is ═O, —OR6, —NR6, —H, —NC(O)R6, —OC(O)R6 or —OC(O)NR6
• R3 is H, —R7, —C(O)R7 or —C(O)NHR7 or a cyclic moiety bridging C28 and C30
• RC28 is halo or —OR3
• RC29 is H, OH or OMe
• where each substituent is present in either stereochemical orientation unless otherwise indicated, and where R1, R4, R5, R6, R7, R9, and R10 are independently selected from H, aliphatic, heteroaliphatic, aryl or heteroaryl;
• R8 is H, halo, —CN, ═O, —OH, —NR9R10, OSO2CF3, OSO2F, OSO2R4′, OCOR4′, OCONR4′R5′, or OCON(OR4′)R5′;
• in which one or both of RC13 and RC28 is a halo substituent;
• both RC24 and RC30 are other than ═O;
• one of RC7a and RC7b is H and the other is phenyl, di- or tri-substituted phenyl or a mono- or di-substituted heterocyclic moiety;
• n is 1;
• and/or moiety “a” is other than
• as a substantially pure stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable derivative thereof.