Derivatives of magainin and methods of production thereof

Abstract

The present invention provides a Magainin derivative peptide and method of production thereof. Also provided is a pharmaceutically composition comprising said Magainin derivative peptide and pharmaceutically acceptable carrier and/or pharmaceutically compatible binding agents. The Magainin derivative peptide of the present invention having amino acid sequence of the general formula shown as below: Gly-Ile-Gly-Lys-Phe-Leu-His-Ser-Ala-Lys-Lys-Phe-Gly-Lys-Ala-Phe-Val-Gly-Glu-Ile-X-Asn-Y-Z-OH in which: X is an amino acid residue selected from the group consisting of Met, Ile and Leu; Y is an amino acid residue selected from the group consisting of Ser, Lys, Ile, Arg and Leu; and Z is Arg.

1 - 6 . (canceled)  
     
     
         7 . An isolated DNA sequence comprising a DNA sequence encoding a peptide of Magainin derivative having amino acid sequence of the general formula shown as below:  
         Gly-Ile-Gly-Lys-Phe-Leu-His-Ser-Ala-Lys-Lys-Phe-Gly-Lys-Ala-Phe-Val-Gly-Glu-Ile-X-Asn-Y-Z-OH  in which:    X is an amino acid selected from the group consisting of Met, Ile and Leu;    Y is an amino acid selected from the group consisting of Ser, Lys, Ile, Leu and Arg, and; Z is Arg.    
     
     
         8 . An isolated DNA sequence comprising a DNA sequence encoding the fusion peptide of at least two tandemly linked peptides of Magainin derivative having amino acid sequence of the general formula shown as below:  
         Gly-Ile-Gly-Lys-Phe-Leu-His-Ser-Ala-Lys-Lys-Phe-Gly-Lys-Ala-Phe-Val-Gly-Glu-Ile-X-Asn-Y-Z-OH  in which:    X is an amino acid selected from the group consisting of Met, Ile and Leu;    Y is an amino acid selected from the group consisting of Ser, Lys, Ile, Leu and Arg, and, Z is Arg.    
     
     
         9 . An expression vector comprising the isolated DNA sequence of  claim 7 .  
     
     
         10 . A transformed host cell comprising the expression vector of  claim 9 .  
     
     
         11 . The transformed host cell of  claim 10 , wherein said host cell is selected from the group consisting of bacterial and mammalian host cells.  
     
     
         12 . A method for producing the Magainin derivative peptide of claim  1 , said method comprising: 
 a. constructing an expression vector comprising at least two tandemly linked DNA sequences encoding said Magainin derivative peptide of claim  1  and a promoter sequence, wherein said promoter sequence is capable of driving the expression of said tandemly linked DNA sequences encoding said Magainin derivative peptide;    b. expressing said expression vector in a host cell to produce a fusion protein comprising at least two tandemly linked Magainin derivative peptides; and    c. cleaving said fusion protein into separate Magainin derivative peptides with a cleavage reagent.    
     
     
         13 . The method of  claim 12 , wherein said fusion protein was cleaved by alkaline protease.  
     
     
         14 . The method of  claim 13 , wherein said alkaline protease is trypsin.  
     
     
         15 . The method of  claim 14 , wherein the Lys residue of said Magainin derivative sequence was protected by acetylation.  
     
     
         16 - 19 . (canceled)  
     
     
         20 . A method for the treatment of a patient having need of a Magainin derivative comprising: 
 administering to said patient a therapeutically effective amount of the peptide of claim  1 .    
     
     
         21 . The method of  claim 20 , wherein said therapeutically effective amount of the polypeptides is administered by providing to the patient DNA encoding said peptide and expressing said peptide in vivo.  
     
     
         22 . The method of  claim 20  comprising: administrating to said patient a therapeutically effective amount of the peptide of claim  3 .  
     
     
         23 . The method of  claim 22  comprising: administrating to said patient a therapeutically effective amount of the peptide of claim  4 .  
     
     
         24 . The method of  claim 23  comprising: administrating to said patient a therapeutically effective amount of the peptide of claim  5 .