Chimeric proteins with cell-targeting specificity and apoptosis-inducing activities

Abstract

The present invention relates to chimeric proteins with cell-targeting specificity and apoptosis-inducing activities. In particular, the invention is illustrated by examples of chimeric proteins including recombinant chimeric proteins between a gonadoptropin releasing hormone (GnRH) and an apoptosis-inducing moiety, such as a pro-apoptotic member of Bcl-2 family, such as Bik, Bax and Bak, or caspases, such as caspase-3. The chimeric proteins specifically target cells expressing a GnRH-binding site, such as adenocarcinoma cells, and induce cell-specific apoptosis.

1 - 36 . (canceled) 
     
     
         37 . A chimeric protein comprising a cell-specific targeting moiety and an apoptosis-inducing moiety, wherein the cell-specific targeting moiety is selected from the group consisting of gonadotropin releasing hormone and an analog thereof, and the apoptosis-inducing moiety is a protein of human origin and induces apoptosis upon entry into a target cell. 
     
     
         38 . (canceled) 
     
     
         39 . The chimeric protein according to  claim 37 , wherein the apoptosis-inducing moiety is DNA fragmentation factor, DFF40. 
     
     
         40 . The chimeric protein according to  claim 37 , wherein the apoptosis-inducing moiety is a pro-apoptotic member of the Bc1-2 family or a domain thereof having an apoptosis-inducing activity. 
     
     
         41 . The chimeric protein according to  claim 40 , wherein the pro-apoptotic member of the Bc1-2 family is Bik, Bax or Bak. 
     
     
         42 . The chimeric protein according to  claim 37 , wherein the apoptosis-inducing moiety is a caspase. 
     
     
         43 . The chimeric protein according to  claim 42 , wherein the caspase is caspase-3. 
     
     
         44 . The chimeric protein according to  claim 37  which is produced by a recombinant DNA method. 
     
     
         45 . The chimeric protein according to  claim 37  which is produced by a chemical conjugation method. 
     
     
         46 . The chimeric protein according to  claim 37 , wherein the two moieties are connected by a polylinker. 
     
     
         47 . A pharmaceutical composition comprising a chimeric protein according to  claim 37  and a pharmaceutically acceptable carrier. 
     
     
         48 . (canceled) 
     
     
         49 . A pharmaceutical composition comprising a chimeric protein according to  claim 39  and a pharmaceutically acceptable carrier. 
     
     
         50 . A pharmaceutical composition comprising a chimeric protein according to  claim 40  and a pharmaceutically acceptable carrier. 
     
     
         51 . A pharmaceutical composition comprising a chimeric protein according to  claim 41  and a pharmaceutically acceptable carrier. 
     
     
         52 . A pharmaceutical composition comprising a chimeric protein according to  claim 42  and a pharmaceutically acceptable carrier. 
     
     
         53 . A pharmaceutical composition comprising a chimeric protein according to  claim 43  and a pharmaceutically acceptable carrier. 
     
     
         54 . A method of treating a cancer cell comprising administering a therapeutically effective amount of chimeric protein comprising a cell-specific targeting moiety and an apoptosis-inducing moiety, wherein the cell-specific targeting moiety binds GnRH binding site. 
     
     
         55 . The method according to  claim 54 , wherein the cell-specific targeting moiety is GnRH or an analog thereof. 
     
     
         56 . The method according to  claim 54 , wherein the apoptosis inducing moiety is DFF40. 
     
     
         57 . The method according to  claim 54 , wherein the apoptosis inducing moiety is a pro-apoptotic member of Bc1-2 family or a domain thereof having an apoptosis-inducing activity. 
     
     
         58 . The method according to  claim 57 , wherein the pro-apoptotic member of Bc1-2 family is Bik, Bax, or Bak. 
     
     
         59 . The method according to  claim 54 , wherein the apoptosis-inducing moiety is a caspase. 
     
     
         60 . The method according to  claim 59 , wherein the caspase is Caspase3 or proCaspase3. 
     
     
         61 . The method according to  claim 54 , wherein the cancer cell is an adenocarcinoma cell. 
     
     
         62 . The chimeric protein according to  claim 37 , wherein the apoptosis-inducing moiety is cytochrome c. 
     
     
         63 . A pharmaceutical composition comprising a chimeric protein according to  claim 62  and a pharmaceutically acceptable carrier. 
     
     
         64 . The method according to  claim 54 , wherein the apoptosis-inducing moiety is cytochrome c. 
     
     
         65 . The chimeric protein of  claim 37 , wherein the chimeric protein cleaves cellular substrates selected from the group consisting of poly(ADP-ribose) polymerase and lamins.