Patent ID: 6649595
Filing Date: 2003-11-18
Classification: A61K,C07D,C07K,C12N

Abstract:
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 FK506-binding protein (FKBP) domain and at least one protein domain heterologous thereto, wherein the FKBP domain comprises a naturally occurring FKBP or a variant thereof in which up to 10 amino acid residues have been deleted, inserted, or replaced with substitute amino acids; (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 FKBP:rapamycin binding (FRB) domain and at least one domain heterologous thereto, wherein the FRB domain comprises a peptide sequence selected from a naturally occurring FRB domain, or a variant thereof in which up to 10 amino acid residues have been deleted, inserted, or replaced with substitute amino acids; 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 has an immunosup-pressive effect less than 0.01 times that of rapamycin and comprises the substructure of formula I: bearing one or more optional substituents, optionally unsaturated at one or more carbonâ€”carbon bonds spanning carbons 1 through 8, as a substantially pure stereoisomer or mixture of stereoisomers, or a pharmaceutically acceptable derivative thereof.