Peptidic antagonists of class iii semaphorins/neuropilins complexes

Abstract

The present invention concern a peptidic antagonist of class III semaphorins/neuropilins complexes comprising an amino acid sequence, which is derived from the transmembrane domain of a protein selected in the group consisting of neuropilin-1, neuropilin-2, plexin-A1, plexin-A2, plexin-A3, plexin-A4, Nr-CAM, L1-CAM, integrin Beta 1 and integrin beta 2, and including at least a GxxxG motif, eventually fused to an heterologous sequence; a nucleic acid encoding for said peptidic antagonist, a pharmaceutical composition comprising such a peptidic antagonist or a nucleic acid encoding thereof and uses thereof.

1 . A peptidic antagonist of class III semaphorins/neuropilins complexes consisting of:
 a transmembrane domain of a protein selected from the group consisting of neuropilin-1, neuropilin-2, plexin-A1, plexin-A2, plexin-A3, plexin-A4, Nr-CAM, L1-CAM, integrin beta 1 and integrin beta 2, wherein the transmembrane domain is optionally fused to a heterologous sequence, or   an amino acid sequence of less than 50 amino acid long and more than 14 amino acid long having an identity of more than 80% with a transmembrane domain of a protein selected from the group consisting of neuropilin-1, neuropilin-2, plexin-A1, plexin-A2, plexin-A3, plexin-A4, Nr-CAM, L1-CAM, integrin beta 1 and integrin beta 2 and including at least one GxxxG motif, wherein the amino acid sequence of less than 50 amino acid long and more than 14 amino acid long retains the biological properties of the transmembrane domain and wherein the amino acid sequence of less than 50 amino acid long and more than 14 amino acid long is optionally fused to the heterologous sequence.   
     
     
         2 . The peptidic antagonist of  claim 1 , wherein the amino acid sequence of less than 50 amino acid long and more than 14 amino acid long has an identity of more than 85% with the transmembrane domain amino acid sequence. 
     
     
         3 . The peptidic antagonist of  claim 1 , wherein the amino acid sequence of less than 50 amino acid long and more than 14 amino acid long has an identity of more than 90% with the transmembrane domain amino acid sequence. 
     
     
         4 . The peptidic antagonist of  claim 1 , wherein the transmembrane domain has the amino acid sequence of the transmembrane domain of human neuropilin-1 as set forth in SEQ ID NO: 1, of human neuropilin-2 as set forth in SEQ ID NO: 2, of human plexin A1 as set forth in SEQ ID NO: 3, of human plexin A2 as set forth in SEQ ID NO: 11, of human plexin A3 as set forth in SEQ ID NO: 12, of human plexin A4 as set forth in SEQ ID NO: 6, of human Nr-CAM as set forth in SEQ ID NO: 7, of human L1-CAM as set forth in SEQ ID NO: 13, of human integrin beta 1 as set forth in SEQ ID NO: 9 or of human integrin beta 2 as set forth in SEQ ID NO: 14. 
     
     
         5 . The peptidic antagonist of  claim 1 , wherein the heterologous sequence allows at least one of: specific cellular location of the peptidic antagonist or improved purification of the peptidic antagonist. 
     
     
         6 . A nucleic acid encoding the peptidic antagonist according of  claim 1 . 
     
     
         7 . A vector comprising the nucleic acid of  claim 6 . 
     
     
         8 . A composition comprising the peptidic antagonist of  claim 1  and at least one pharmaceutically acceptable carrier. 
     
     
         9 . The composition of  claim 8 , wherein the composition comprises a concentration of more than 10 −12  M of the peptidic antagonist. 
     
     
         10 . The composition of  claim 8 , wherein the composition comprises a concentration of more than 10 −11  M of peptidic antagonist. 
     
     
         11 . A composition comprising the nucleic acid of  claim 6  and at least one pharmaceutically acceptable carrier. 
     
     
         12 . A composition comprising the vector of  claim 7  and at least one pharmaceutically acceptable carrier. 
     
     
         13 . A method for treatment of a subject suffering from a disease associated with class III semaphorins/neuropilins complexes signal transduction pathways,
 comprising administering to the subject a pharmaceutical composition comprising a peptidic antagonist of class III semaphorins/neuropilins complexes,   the peptidic antagonist
 consisting of a transmembrane domain of a protein selected from the group consisting of plexin-A2, plexin-A3, plexin-A4, Nr-CAM, L1-CAM, and integrin beta 2, or 
 consisting of a transmembrane domain of a protein selected from the group consisting of plexin-A2, plexin-A3, plexin-A4, Nr-CAM, L1-CAM, and integrin beta 2, wherein the transmembrane domain is fused to a heterologous sequence, 
   wherein the disease is selected from the group consisting of diseases associated with excessive angiogenesis and cancers.   
     
     
         14 . The method of  claim 13 , wherein administration of the pharmaceutical composition releases a concentration of the peptidic antagonist of more than 10 −12  M. 
     
     
         15 . The method of  claim 13 , wherein the transmembrane domain has an amino acid sequence of the transmembrane domain of human plexin A2 as set forth in SEQ ID NO: 11, of human plexin A3 as set forth in SEQ ID NO: 12, of human plexin A4 as set forth in SEQ ID NO: 6, of human Nr-CAM as set forth in SEQ ID NO: 7, of human L1-CAM as set forth in SEQ ID NO: 13, or of human integrin beta 2 as set forth in SEQ ID NO: 14. 
     
     
         16 . The method of  claim 13 , wherein the heterologous sequence allows at least one of: specific cellular location of the peptidic antagonist or improved purification yield of the peptidic antagonist.