Bifunctional stapled polypeptides and uses thereof

Abstract

The invention relates to bifunctional stapled or stitched peptides comprising a targeting domain, a linker moiety, and an effector domain, that can be used to tether, or to bring into close proximity, at least two cellular entities (e.g., proteins). Certain aspects relate to bifunctional stapled or stitched peptides that bind to an effector biomolecule through the effector domain and bind to a target biomolecule through the targeting domain. Polypeptides and/or polypeptide complexes that are tethered by the bifunctional stapled or stitched peptides of the invention, where the effector polypeptide bound to the effector domain of the bifunctional stapled or stitched peptide modifies or alters the target polypeptide bound to the targeting domain of the bifunctional peptide. Uses of the inventive bifunctional stapled or stitched peptides including methods for treatment of disease (e.g., cancer, inflammatory diseases) are also provided.

1 - 111 . (canceled) 
     
     
         112 . A bifunctional stapled peptide comprising an effector domain selected from: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 36) 
                 
                     
                   Ac-LSQETFS*LWK*LPE-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 37) 
                 
                     
                   Ac-QSQQTF#NLWRKK*QN-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 38) 
                 
                     
                   Ac-QSQQTF*NLW*KKQN-CONH 2 ; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 39) 
                 
                     
                   Ac-LSQNTFS*LWK*LPQ-CONH 2 , 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       where “*” is the non-natural amino acid S5, and “#” is the non-natural amino acid R8. 
     
     
         113 . A bifunctional stapled peptide comprising an effector domain selected from: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 67) 
                 
                     
                   Ac-VRIVINIQMLLEA*DYL*RRER-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 68) 
                 
                     
                   Ac-VRIVINIQM*LEA*DYLERRER-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 69) 
                 
                     
                   Ac-VRIVINIQML#EAADYL*RRER-CONH 2 ; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 70) 
                 
                     
                   Ac-VRIVI*IQM*LEAADYLERRER-CONH 2 , 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       where “*” is the non-natural amino acid S5, and “#” is the non-natural amino acid R8. 
     
     
         114 . A bifunctional stapled peptide comprising an effector domain selected from: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 52) 
                 
                     
                   Ac-KEKRIKELEL*LMS*ENELKG-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 53) 
                 
                     
                   Ac-KEKRIK*LEL*LMSTENELKG-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 54) 
                 
                     
                   Ac-KE*RIK*LELLLMSTENELKG-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 55) 
                 
                     
                   Ac-KEKRIK#LELLLM*TENELKG-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 73) 
                 
                     
                   K*KRI*ELELLLMSTENELKG; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 74) 
                 
                     
                   K*KRI*RLELLLMSTENELKG; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 75) 
                 
                     
                   KE*RIK*LELLLMSTENELKG; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 76) 
                 
                     
                   KR*RIK*LELLLMSTENELKG; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 77) 
                 
                     
                   KE*RIKELE*LLMSTENELKG; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 78) 
                 
                     
                   KE*RIKRLE*LLMSTENELKG; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 79) 
                 
                     
                   KR*RIKELE*LLMSTENELKG; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 80) 
                 
                     
                   KEKRIKELELLLMSTE*ELK*; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 56) 
                 
                     
                   Ac-*ILP*DIMDFVLKNTP-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 57) 
                 
                     
                   Ac-ILP*DEVI*FVLKNTP-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 58) 
                 
                     
                   Ac-ILPSDIM*FVL*NTP-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 59) 
                 
                     
                   Ac-ILPSDIMDFV*KNT*-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 60) 
                 
                     
                   Ac-#ILPSDI*DFVLKNTP-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 81) 
                 
                     
                   ILP*DEVI*FVLKNT; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 82) 
                 
                     
                   ILP*RIM*FVLKNT; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 83) 
                 
                     
                   ILPSDIM*FVL*NT; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 84) 
                 
                     
                   ILPSRIM*FVL*NT; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 61) 
                 
                     
                   Ac-ILSRRPSY*KIL*DLSSDAPG-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 62) 
                 
                     
                   Ac-ILSRRPSYRKIL*DLS*DAPG-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 63) 
                 
                     
                   Ac-ILSR*PSY*KILNDLSSDAPG-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 64) 
                 
                     
                   Ac-ILSRRPSYR*ILN*LSSDAPG-CONH 2 ; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 65) 
                 
                     
                   Ac-ILSRRP#YRKILN*LSSDAPG-CONH 2 ; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 66) 
                 
                     
                   Ac-ILSRRPSYRKILNDLSSDAPG-CONH 2 , 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       where “*” is the non-natural amino acid S5, and “#” is the non-natural amino acid R8. 
     
     
         115 . A pharmaceutical composition comprising a peptide of  claim 112  or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. 
     
     
         116 . A pharmaceutical composition comprising a peptide of  claim 113  or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. 
     
     
         117 . A pharmaceutical composition comprising a peptide of  claim 114  or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. 
     
     
         118 . A method of treating a disease, disorder, or condition in a subject, comprising administering a therapeutically effective amount of a peptide of  claim 112  to a subject in need thereof. 
     
     
         119 . The method of  claim 118 , wherein the disease, disorder, or condition is cancer, hematopoietic neoplastic disorders, proliferative breast disease, proliferative disorders of the lung, proliferative disorders of the colon, proliferative disorders of the liver, and proliferative disorders of the ovary. 
     
     
         120 . A method of treating a disease, disorder, or condition in a subject, comprising administering a therapeutically effective amount of a peptide of  claim 113  to a subject in need thereof. 
     
     
         121 . The method of  claim 120 , wherein the disease, disorder, or condition is cancer, hematopoietic neoplastic disorders, proliferative breast disease, proliferative disorders of the lung, proliferative disorders of the colon, proliferative disorders of the liver, and proliferative disorders of the ovary. 
     
     
         122 . A method of treating a disease, disorder, or condition in a subject, comprising administering a therapeutically effective amount of a peptide of  claim 114  to a subject in need thereof. 
     
     
         123 . The method of  claim 122 , wherein the disease, disorder, or condition is cancer, hematopoietic neoplastic disorders, proliferative breast disease, proliferative disorders of the lung, proliferative disorders of the colon, proliferative disorders of the liver, and proliferative disorders of the ovary.