Process for identifying a ligand that binds to the nep binding site for the smr1 pentapeptide

Abstract

The invention relates to the therapeutic use of a SMR1-peptide or a pharmaceutically active amount of said SMR1-peptide, for the preparation of a therapeutic composition for preventing or treating diseases wherein a modulation of the activity of a membrane metallopeptidase, notably a membrane-zinc metallopeptidase, is sought, in a mammal, specifically in a human.

1 - 36 . (canceled) 
     
     
         37 . A process for screening for a ligand molecule that specifically binds to the NEP binding site for the QHNPR (SEQ ID NO: 2) pentapeptide, comprising:
 a) preparing a cell culture or preparing an organ specimen or a tissue sample containing NEP binding sites for the QHNPR (SEQ ID NO: 2) pentapeptide;   b) adding a candidate molecule to be tested in competition with a half-saturating concentration of the labeled pentapeptide;   c) incubating the cell culture, organ specimen or tissue sample of a) in the presence of the labeled candidate molecule for a time and under conditions sufficient for the specific binding to take place; and   d) quantifying the label specifically bound to the cell culture, organ specimen or tissue sample in the presence of various concentrations of said candidate molecule.   
     
     
         38 . The process of  claim 37 , further comprising
 determining the relative affinity of the candidate ligand molecules that specifically bind to the NEP binding site for the QHNPR (SEQ ID NO: 2) pentapeptide by   e) comparing the affinity of each candidate molecule quantified in d) to the affinity of the other candidate molecules quantified.   
     
     
         39 . The method of  claim 37 , wherein said candidate ligand molecule comprises SEQ ID NO: 10 or a peptidomimetic thereof. 
     
     
         40 . A process for determining the affinity of a ligand molecule that specifically binds to the NEP binding site for the QHNPR (SEQ ID NO: 2) pentapeptide, comprising:
 a) preparing a cell culture or preparing an organ specimen or a tissue sample containing NEP binding sites for the QHNPR (SEQ ID NO: 2) pentapeptide;   b) adding a candidate ligand molecule which has previously been labeled with a radioactive or a nonradioactive label;   c) incubating the cell culture, organ specimen or tissue sample of a) in the presence of the labeled candidate ligand molecule for a time sufficient and under conditions for the specific binding to take place; and   d) quantifying the label specifically bound to the cell culture, organ specimen or tissue sample in the presence of various concentrations of labeled ligand molecule.   
     
     
         41 . The method of  claim 40 , wherein said candidate ligand molecule comprises SEQ ID NO: 10 or a peptidomimetic thereof. 
     
     
         42 . An isolated or purified molecule comprising SEQ ID NO: 10 or a peptidomimetic thereof. 
     
     
         43 . The isolated or purified molecule of  claim 42 , which comprises SEQ ID NO: 10. 
     
     
         44 . The isolated or purified molecule of  claim 42 , which is a peptidomimetic of SEQ ID NO: 10. 
     
     
         45 . The isolated or purified molecule of  claim 42 , which inhibits the normal interaction between an SMR1 peptide and a metalloproteinase. 
     
     
         46 . The isolated or purified molecule of  claim 45 , wherein said metalloproteinase is a zinc metalloproteinase. 
     
     
         47 . The isolated or purified molecule of  claim 45 , wherein said metalloproteinase is NEP. 
     
     
         48 . The isolated or purified molecule of  claim 42 , which increases metalloproteinase activity. 
     
     
         49 . The isolated or purified molecule of  claim 42 , which decreases metalloproteinase activity. 
     
     
         50 . A molecular complex comprising:
 an SMR-1 peptide, and   an NEP receptor, or a SMR1-peptide binding site of an NEP receptor.