Photocleavable Chemical Inducers of Dimerization (CID) and Methods of Use

Abstract

The instant invention provides photolysable compounds, and their use in reversible chemical induced dimerization and light-induced regulation of proteins.

1 . A compound represented by the formula R 1 —Y—R 2 , wherein
 R 1  is a ligand capable of selectively binding to a first receptor; 
 R 2  is a ligand capable of selectively binding to a second receptor; and 
 Y is a linker providing a covalent linkage between R 1  and R 2 , wherein Y is photocleavable. 
 
     
     
         2 . The compound of  claim 1 , wherein R 1  is a ligand capable of selectively binding to dihydrofolate reductase (DHFR). 
     
     
         3 . The compound of  claim 2 , wherein the ligand capable of selectively binding to dihydrofolate reductase is trimethoprim (TMP) or a derivative or analog thereof, or methotrexate or a derivative or analog thereof. 
     
     
         4 . The compound of  claim 2 , wherein R 2  is a ligand capable of selectively binding to FK506 Binding Protein (FKBP). 
     
     
         5 . The compound of  claim 4 , wherein the ligand capable of selectively binding to FK506 Binding Protein (FKBP) is a synthetic ligand for FKBP (SLF) or a derivative or analog thereof. 
     
     
         6 . The compound of  claim 1 , wherein Y is cleavable by exposure to light at a wavelength of about 405 nm. 
     
     
         7 . The compound of  claim 6 , wherein Y comprises a dimethoxynitrobenzyl (DMNB) moiety or a derivative or analog thereof. 
     
     
         8 . A complex comprising a compound of  claim 1  complexed to (i) a first protein domain comprising a first binding domain and (ii) a second protein domain comprising a second binding domain. 
     
     
         9 . A method of regulating activity of a protein, the method comprising:
 (a) providing a complex comprising   (i) a first protein domain comprising a first binding domain;   (ii) a second protein domain comprising a second binding domain; and   (iii) a compound of  claim 1 ,   wherein the compound of  claim 1  is bound to the first protein domain and the second protein domain;   (b) cleaving the compound of  claim 1  by exposure to light, thereby cleaving the complex and regulating activity of the protein.   
     
     
         10 . The method of  claim 9 , wherein the first binding domain is dihydrofolate reductase (DHFR). 
     
     
         11 . The method of  claim 10 , wherein the DHFR is  E. coli  DHFR. 
     
     
         12 . The method of  claim 9 , wherein the second binding domain is FK506 binding protein (FKBP). 
     
     
         13 . The method of  claim 9 , wherein the first protein domain comprises one domain of an active protein. 
     
     
         14 . The method of  claim 9 , wherein the second protein domain comprises another domain of an active protein. 
     
     
         15 . The method of  claim 9 , wherein the first protein domain and the second protein domain together comprise a functional protein. 
     
     
         16 . The method of  claim 9 , wherein regulating the activity of a protein comprises inactivating the protein. 
     
     
         17 . The method of  claim 9 , wherein regulating the activity of a protein comprises activating the protein. 
     
     
         18 . A method of regulating the activity of a protein in a cell, the method comprising:
 (a) providing a cell that expresses a first protein domain comprising a first binding domain and a second protein domain that comprises a second binding domain;   (b) providing the compound of  claim 1  that dimerizes the first and second protein domains; and   (c) cleaving the compound of  claim 1  by exposure to light, thereby cleaving the complex and regulating activity of the protein.   
     
     
         19 . The method of  claim 18 , wherein dimerization of the first and second protein domains produces an active protein. 
     
     
         20 . The method of  claim 18 , wherein dimerization of the first and second protein domains inactivates the protein.