Peptides derived from fibronectin with improved bioactivity and reduced susceptibility to neutrophil elastase degradation

Abstract

Polypeptides derived from fibronectin are presented that are neutrophil elastase-resistant and can bind to growth factors and/or enhance growth factor activity. These polypeptides are useful for enhancing wound healing in a patient.

1 . A linear or branched multimeric polypeptide or a cyclized form thereof, comprising two or more independently determined sequences, each independently determined sequence consisting of Formula I: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 1) 
                 
                     
                   (I): H-X 1 -X 2 -K-Y-X 3 -X 4 -R-W-R-P-K-X 5 -X 6 -X 7   
                 
             
                
                
               
            
           
         
         wherein X 1  is I or G or L,
 X 2  is S or G, 
 X 3  is I or G or L, 
 X 4  is L or G, 
 X 5  is N or G, 
 X 6  is absent or S, and 
 X 7  is absent or V; and 
 
         wherein no two consecutive amino acids in the first 13 amino acids of each independently determined sequence of Formula I differ from the sequence HISKYILRWRPKN (SEQ ID NO:9). 
       
     
     
         2 . The linear or branched multimeric polypeptide or cyclized form thereof of  claim 1 , with a linker between each independently determined sequence of Formula I. 
     
     
         3 . The linear or branched multimeric polypeptide or cyclized form thereof of  claim 1 , wherein the polypeptide is substituted with at least one of a lower acyl group at the N-terminus, a substituted or unsubstituted lower alkyl-, alkenyl-, alkynyl- or haloalkyl-amine group at the C-terminus and polyethylene glycol. 
     
     
         4 . The linear or branched multimeric polypeptide or cyclized form thereof of  claim 1 , wherein each independently determined sequence of Formula I consists of HISKYILRWRPKNSV (SEQ ID NO:10), HIGKYGLRWRPKNSV (SEQ ID NO:11), HIGKYGLRWRPKGSV (SEQ ID NO:12), HGSKYGLRWRPKNSV (SEQ ID NO:13), HIGKYIGRWRPKNSV (SEQ ID NO:14), HGSKYIGRWRPKNSV (SEQ ID NO:15), HGSKYIGRWRPKGSV (SEQ ID NO:16) and any combination thereof. 
     
     
         5 . The linear or branched multimeric polypeptide or cyclized form thereof of  claim 4 , wherein each independently determined sequence of Formula I consists of 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 12) 
                 
                     
                   HIGKYGLRWRPKGSV. 
                 
             
                
                
               
            
           
         
       
     
     
         6 . A composition comprising:
 the linear or branched multimeric polypeptide or cyclized form thereof of  claim 1 , and;   a pharmaceutically acceptable excipient, carrier or diluent, wherein the composition is suitable for a route of administration selected from injection, intravenous administration and topical administration to a patient.   
     
     
         7 . A method of treating a patient with a wound selected from the group consisting of a surgical incision or extirpation, a traumatic injury, a thermal burn, a chemical burn, a lesion or ulceration of the patient's skin, mucosa, connective tissue, fascia, ligament, tendon, cartilage, nerve or muscle and a wound to the patient's bone, the method comprising:
 administering to the patient a therapeutically effective amount of a composition comprising a linear or branched multimeric polypeptide or cyclized form thereof comprising two or more independently determined sequences, each independently determined sequence consisting of Formula I:   
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 1) 
                 
                     
                   (I): H-X 1 -X 2 -K-Y-X 3 -X 4 -R-W-R-P-K-X 5 -X 6 -X 7   
                 
             
                
                
               
            
           
         
         wherein X 1  is I or G or L,
 X 2  is S or G, 
 X 3  is I or G or L, 
 X 4  is L or G, 
 X 5  is N or G, 
 X 6  is absent or S, and 
 X 7  is absent or V; and 
 
         wherein no two consecutive amino acids in the first 13 amino acids each independently determined sequence of Formula I differ from the sequence 
       
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 9) 
                 
                     
                   HISKYILRWRPKN. 
                 
             
                
                
               
            
           
         
       
     
     
         8 . The method of  claim 7 , with a linker between each independently determined sequence of Formula I. 
     
     
         9 . The method of  claim 7 , wherein the polypeptide is substituted with at least one of a lower acyl group at the N-terminus, a substituted or unsubstituted lower alkyl-, alkenyl-, alkynyl- or haloalkyl-amine group at the C-terminus and polyethylene glycol. 
     
     
         10 . The method of  claim 7 , wherein each independently determined sequence of Formula I consists of HISKYILRWRPKNSV (SEQ ID NO:10), HIGKYGLRWRPKNSV (SEQ ID NO:11), HIGKYGLRWRPKGSV (SEQ ID NO:12), HGSKYGLRWRPKNSV (SEQ ID NO:13), HIGKYIGRWRPKNSV (SEQ ID NO:14), HGSKYIGRWRPKNSV (SEQ ID NO:15), HGSKYIGRWRPKGSV (SEQ ID NO:16) and any combination thereof. 
     
     
         11 . The method of  claim 10 , wherein each independently determined sequence of Formula I consists of HIGKYGLRWRPKGSV (SEQ ID NO:12).