Treatment of wounds

Abstract

An isolated protein, for use in treatment of wounds, is characterized in that it is secreted by the organism Lucilia sericata and it exhibits proteolytic activity against FITC-casein at a pH of 8.0 to 8.5. The protein exhibits proteolytic activity against Tosyl-Gly-Pro-Arg-AMC but not against Suc-Ala-Ala-Phe-AMC, and its proteolytic activity against FITC-casein and Tosyl-Gly-Pro-Arg-AMC is inhibited by the serine proteinase inhibitors PMSF and AMPSF. The protein is also bound by immobilized aminobenzamidine.

1 . (canceled)  
     
     
         2 . The method of  claim 6 , wherein the isolated protein has a molecular weight of approximately 25 kDa.  
     
     
         3 . (canceled)  
     
     
         4 . The method of  claim 6 , wherein the isolated protein is secreted by an insect in its larval stage.  
     
     
         5 . (canceled)  
     
     
         6 . A method for treating a wound to promote healing thereof in a human or non-human mammal, which comprises applying to the wound a therapeutically effective amount of: 
 (a) an isolated protein characterized in that: 
 i) it is secreted by the organism  Lucilia sericata;    
 ii) it exhibits optimum proteolytic activity against FITC-casein at a pH of 8.0 to 8.5;  
 iii) it exhibits proteolytic activity against Tosyl-Gly-Pro-Arg-AMC but not against Suc-Ala-Ala-Phe-AMC;  
 iv) its proteolytic activity against FITC-casein and Tosyl-Gly-Pro-Arg-AMC is inhibited by the serine proteinase inhibitors PMSF and APMSF; and  
 v) it is bound by immobilized aminobenzamidine; or  
   (b) one or more peptides selected from the group consisting of Ser-Phe-Leu-Leu-Arg-Asn (SEQ ID NO: 1); Ser-Leu-Ile-Gly-Lys-Val (SEQ ID NO: 2); Thr-Phe-Arg-Gly-Ala-Pro (SEQ ID NO: 3); Gly-Tyr-Pro-Gly-Gln-Val ((SEQ ID NO: 4); a peptide having as an N-terminal sequence Ser-Phe-Leu-Leu-Arg-Asn (SEQ ID NO: 1), Ser-Leu-Ile-Gly-Lys-Val (SEQ ID NO: 2), Thr-Phe-Arg-Gly-Ala-Pro (SEQ ID NO: 3), or Gly-Tyr-Pro-Gly-Gln-Val (SEQ ID NO: 4); and analogues thereof which are protected against aminopeptidase activity.    
     
     
         7 . The method of  claim 6  wherein the isolated protein or peptide is applied as a sterile composition comprising a therapeutically effective amount of the isolated protein or peptide and a sterile carrier therefore.  
     
     
         8 . (canceled)  
     
     
         9 . (canceled)  
     
     
         10 . An isolated peptide selected from the group consisting of 
 Ser-Phe-Leu-Leu-Arg-Asn (SEQ ID NO: 1); Ser-Leu-Ile-Gly-Lys-Val (SEQ ID NO: 2); Thr-Phe-Arg-Gly-Ala-Pro (SEQ ID NO: 3); Gly-Tyr-Pro-Gly-Gln-Val (SEQ ID NO: 4); a peptide having as an N-terminal sequence Ser-Phe-Leu-Leu-Arg-Asn (SEQ ID NO: 2), Ser-Leu-Ile-Gly-Lys-Val (SEQ ID NO: 2), Thr-Phe-Arg-Gly-Ala-Pro (SEQ ID NO:3), or Gly-Tyr-Pro-Gly-Gln-Val (SEQ ID NO: 4); and analogues thereof which are protected against aminopeptidase activity    
     
     
         11 . (canceled)  
     
     
         12 . (canceled)  
     
     
         13 . A dressing for a wound, which comprises a sterile support and isolated protein characterised in that: 
 i) it is secreted by the organism  Lucilia sericata;      ii) it exhibits optimum proteolytic activity against FITC-casein at a pH of 8.0 to 8.5;    iii) it exhibits proteolytic activity against Tosyl-Gly-Pro-Arg-AMC but not against Suc-Ala-Ala-Phe-AMC;    iv) its proteolytic activity against FITC-casein and Tosyl-Gly-Pro-Arg-AMC is inhibited by the serine proteinase inhibitors PMSF and APMSF; and    v) it is bound by immobilised aminobenzamidine.    
     
     
         14 . A dressing for a wound, which comprises a sterile support and at least one peptide according to  claim 10 .  
     
     
         15 . A protein which is protected against aminopeptidase activity, wherein the protein is characterized in that: 
 i) it is secreted by the organism  Lucilia sericata;      ii) it exhibits optimum proteolytic activity against FITC-casein at a pH of 8.0 to 8.5;    iii) it exhibits proteolytic activity against Tosyl-Gly-Pro-Arg-AMC but not against Suc-Ala-Ala-Phe-AMC;    iv) its proteolytic activity against FITC-casein and Tosyl-Gly-Pro-Arg-AMC is inhibited by the serine proteinase inhibitors PMSF and APMSF; and    v) it is bound by immobilized aminobenzamidine.    
     
     
         16 . A protein which is protected against aminopeptidase activity, wherein the protein comprises at least one peptide according to  claim 10 .  
     
     
         17 . The protein of  claim 15  wherein protection against aminopeptidase activity is achieved by amidation at the COOH substitution in the protein using a non-coded anomalous amino acid.  
     
     
         18 . The protein of  claim 15  wherein protection against aminopeptidase activity is achieved by replacing a CO—NH bond by an isostere.  
     
     
         19 . The protein of  claim 15  wherein protection against aminopeptidase activity is achieved by amidation at the COOH substitution in the protein using a non-coded anomalous amino acid and by replacing a CO—NH bond by an isostere.  
     
     
         20 . A composition for the treatment of wounds comprising a therapeutically effective amount of a protein according to  claim 6  and a sterile carrier therefore.  
     
     
         21 . The composition of  claim 20  further comprising a material capable of delivering the protein to a wound in a slow release or controlled release manner.  
     
     
         22 . The composition of  claim 21  wherein the material is poly(lactide-co-glycolide) or particles of PLGA.  
     
     
         23 . The composition of  claim 20  wherein the protein is protected against aminopeptidase activity.  
     
     
         24 . The dressing of  claim 13  wherein the protein is protected against aminopeptidase activity.  
     
     
         25 . The dressing of  claim 13  further comprising slow-release colloidal particles.  
     
     
         26 . The dressing of  claim 13  further comprising sponges.  
     
     
         27 . The dressing of  claim 13  which is over layered by a conventional dressing.