Anti-endotoxic, antimicrobial cationic peptides, the encoding DNA and methods of use thereof

Abstract

A novel class of cationic peptides having antimicrobial activity is provided. Exemplary peptides of the invention include KWKSFIKKLTSAAKKVVTTAKPLALIS (SEQ ID NO:3) and KGWGSFFKKAAHVGKHVGKAALTHYL (SEQ ID NO:15). Also provided are methods for inhibiting the growth of bacteria utilizing the peptides of the invention. Such methods are useful for the treatment of respiratory infections, such as in cystic fibrosis patients. Such methods are further useful for accelerating wound healing.

1 . An isolated antimicrobial peptide having an amino acid sequence selected from the group consisting of:  
       
         
           
                 
                 
                 
               
                     
                 
                   KWKSFIKKLTSAAKKVVTTAKPLALIS; 
                   (SEQ ID NO:3) 
                     
                 
                     
                 
                   KWKSFIKKLTKAAKKVVTTAKKPLIV; 
                   (SEQ ID NO:4) 
                 
                     
                 
                   KWKKFIKSLTKSAAKTVVKTAKKPLV; 
                   (SEQ ID NO:5) 
                 
                     
                 
                   KWKLFKKIGIGAVLKVLKVLTTGLPALKLTLK; 
                   (SEQ ID NO:6) 
                 
                     
                 
                   KLFKKIGIGAVLKVLKVLTTGLPALKLTLK; 
                   (SEQ ID NO:7) 
                 
                     
                 
                   KWKFKKIGIGAVLKVLKVLTTGLPALKLTLK; 
                   (SEQ ID NO:8) 
                 
                     
                 
                   KLWKLFKKIGIGAVLKVLKVLTTGLPALKLTLK; 
                   (SEQ ID NO:9) 
                 
                     
                 
                   KWKSFIKKLTSAAKKVTTAAKPLTK; 
                   (SEQ ID NO:10) 
                 
                     
                 
                   KWKKFIKKIGIGAVLKVLTTGLPALKLTKK; 
                   (SEQ ID NO:11) 
                 
                     
                 
                   KKWKKFIKKIGIGAVLTTPGAKK; 
                   (SEQ ID NO:12) 
                 
                     
                 
                   GWGSFFKKAAHVGKHVGKAALTHYL-NH2; 
                   (SEQ ID NO:14) 
                 
                     
                 
                   KGWGSFFKKAAHVGKHVGKAALTHYL; 
                   (SEQ ID NO:15) 
                 
                     
                 
                   KGWGSFFKKAAHVGKHVGKAALTHYL-NH2; 
                   (SEQ ID NO:16) 
                 
                     
                 
                   ALWKTMLKKAAHVGKHVGKAALTHYL-NH2; 
                   (SEQ ID NO:17) 
                 
                     
                 
                   SIGSAFKKAAHVGKHVGKAALTHYL-NH2; 
                   (SEQ ID NO:18) 
                 
                     
                 
                   GWGSFFKKAAHVGKHVGKAALGAAARRRK; 
                   (SEQ ID NO:19) 
                 
                     
                 
                   ALWKTMLKKAAHVGKHVGKAALGAAARRRK; 
                   (SEQ ID NO:20) 
                 
                   and 
                 
                     
                 
                   SIGSAFKKAAHVGKHVGKAALGAAARRRK; 
                   (SEQ ID NO:21) 
                 
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       and analogs, derivatives, amidated variations and conservative variations thereof.  
     
     
         2 . An isolated polynucleotide which encodes a peptide of  claim 1 .  
     
     
         3 . An isolated polynucleotide which encodes a peptide selected from the group consisting of:  
       
         
           
                 
                 
                 
               
                     
                 
                   KWKSFIKKLTSAAKKVVTTAKPLALIS 
                   (SEQ ID NO:3) 
                     
                 
                     
                 
                   KWKSFIKKLTKAAKKVVTTAKKPLIV 
                   (SEQ ID NO:4) 
                 
                     
                 
                   KWKKFIKSLTKSAAKTVVKTAKKPLIV 
                   (SEQ ID NO:5) 
                 
                     
                 
                   KWKLFKKIGIGAVLKVLKVLTTGLPALKLTLK 
                   (SEQ ID NO:6) 
                 
                     
                 
                   KLFKKIGIGAVLKVLKVLTTGLPALKLTLK 
                   (SEQ ID NO:7) 
                 
                     
                 
                   KWKFKKIGIGAVLKVLKVLTTGLPALKLTLK 
                   (SEQ ID NO:8) 
                 
                     
                 
                   KLWKLFKKIGIGAVLKVLKVLTTGLPALKLTLK 
                   (SEQ ID NO:9) 
                 
                     
                 
                   KWKSFIKKLTSAAKKVTTAAKPLTK 
                   (SEQ ID NO:10) 
                 
                     
                 
                   KWKKFIKKIGIGAVLKVLTTGLPALKLTKK 
                   (SEQ ID NO:11) 
                 
                     
                 
                   KKWKKFIKKIGIGAVLTTPGAKK 
                   (SEQ ID NO:12) 
                 
                     
                 
                   GWGSFFKKAAHVGKHVGKAALTHYL 
                   (SEQ ID NO:14) 
                 
                     
                 
                   KGWGSFFKKAAHVGKHVGKAALTHYL 
                   (SEQ ID NO:15) 
                 
                     
                 
                   ALWKTMLKKAAHVGKHVGKAALTHYL 
                   (SEQ ID NO:17) 
                 
                     
                 
                   SIGSAFKKAAHVGKHVGKAALTHYL 
                   (SEQ ID NO:18) 
                 
                     
                 
                   GWGSFFKKAAHVGKHVGKAALGAAARRRK 
                   (SEQ ID NO:19) 
                 
                     
                 
                   ALWKTMLKKAAHVGKHVGKAALGAAARRRK; 
                   (SEQ ID NO:20) 
                 
                   and 
                 
                     
                 
                   SIGSAFKKAAHVGKHVGKAALGAAARRRK. 
                   (SEQ ID NO:21) 
                 
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         4 . A method of inhibiting the growth of bacteria comprising contacting the bacteria with an inhibiting effective amount of a peptide having an amino acid sequence selected from the group consisting of:  
       
         
           
                 
                 
                 
                 
               
                     
                     
                 
                     
                   GWGSFFKKAAHVGKHVGKAALTHYL-NJ32; 
                   (SEQ ID NO:14) 
                     
                 
                     
                     
                 
                     
                   KGWGSFFKKAAHVGKHVGKAALTHYL; 
                   (SEQ ID NO:15) 
                 
                     
                     
                 
                     
                   KGWGSFFKKAAHVGKHVGKAALTHYL-NH2; 
                   (SEQ ID NO:16) 
                 
                     
                     
                 
                     
                   ALWKTMLKKAAHVGKHVGKAALTHYL-NH2; 
                   (SEQ ID NO:17) 
                 
                     
                     
                 
                     
                   SIGSAFKKAAHVGKHVGKAALTHYL-NH2; 
                   (SEQ ID NO:18) 
                 
                     
                     
                 
                     
                   GWGSFFKKAAHVGKHVGKAALGAAARRRK; 
                   (SEQ ID NO:19) 
                 
                     
                     
                 
                     
                   ALWKTMLKKAAHVGKHVGKAALGAAARRRK; 
                   (SEQ ID NO:20) 
                 
                     
                     
                 
                     
                   SIGSAFKKAAHVGKHVGKAALGAAARRRK; 
                   (SEQ ID NO:21) 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       and analogs, derivatives, amidated variations and conservative variations thereof.  
     
     
         5 . An isolated polynucleotide which encodes a peptide of  claim 4 .  
     
     
         6 . An isolated polynucleotide which encodes a peptide selected from the group consisting of:  
       
         
           
                 
                 
                 
                 
               
                     
                     
                 
                     
                   GWGSFFKKAAHVGKHVGKAALTHYL-NH2; 
                   (SEQ ID NO:14) 
                     
                 
                     
                     
                 
                     
                   KGWGSFFKKAAHVGKHVGKAALTHYL; 
                   (SEQ ID NO:15) 
                 
                     
                     
                 
                     
                   KGWGSFFKKAAHVGKHVGKALTHYL-NH2; 
                   (SEQ ID NO:16) 
                 
                     
                     
                 
                     
                   ALWKTMLKKAAHVGKRVGKAALTHYL-NH2; 
                   (SEQ ID NO:17) 
                 
                     
                     
                 
                     
                   SIGSAFKKAAHVGKHVGKAALTHYL-NH2; 
                   (SEQ ID NO:18) 
                 
                     
                     
                 
                     
                   GWGSFFKKAAHVGKHVGKAALGAAARRRK; 
                   (SEQ ID NO:19) 
                 
                     
                     
                 
                     
                   ALWKTMLKKAAHVGKHVGKAALGAAARRRK; 
                   (SEQ ID NO:20) 
                 
                     
                     
                 
                     
                   SIGSAFKKAAHVGKHVGKAALGAAARRRK. 
                   (SEQ ID NO:21)