Novel x-conotoxin peptides (-ii)

Abstract

An isolated, synthetic or recombinant χ-conotoxin peptide having the ability to inhibit neuronal amine transporter comprising the following sequence of amino acids: Cys Cys Gly Tyr Lys Leu Cys Xaa5 Xaa6 Cys, SEQ ID NO: 3, where Xaa5 and Xaa6 are independently absent or represent any amino acid residue except Cys; or a sequence in which Gly, Tyr, Lys or Leu are subject to conservative amino acid substitution or side chain modification with the proviso that the peptide is not Mr1A, Mr1B, Mar2, CMrV1A, Bn1.5, Mr1.3 or Au1.4; or a salt, ester, amide, prodrug or cyclised derivative thereof.

What is claimed is: 
     
         1 . An isolated, synthetic or recombinant χ-conotoxin peptide having the ability to inhibit neuronal amine transporter comprising the following sequence of amino acids: 
       
         
           
                 
                 
               
                   SEQ ID NO. 3 
                     
                 
                 
                 
                 
               
                     
                   Cys Cys Gly Tyr Lys Leu Cys Xaa5 XaaG Cys 
                     
                 
             
                
               
            
             
                
               
            
           
         
       
       where Xaa5 and Xaa6 are independently absent or represent any amino acid residue except Cys; or a sequence in which Gly, Tyr, Lys or Leu are subject to conservative amino acid substitution or side chain modification; with the proviso that the peptide is not χ-MrIA, χ-MrIB, Mar2, CMrVIA, Bn1.5, Mr1.3 or Au1.4; 
       or a salt, ester, amide, prodrug or cyclised derivative thereof. 
     
     
         2 . The χ-conotoxin peptide according to  claim 1  comprising the following sequence of amino acids:
 Xaa1 Xaa2 Xaa3 Xaa4 Cys Cys Gly Tyr Lys Leu Cys Xaa5 Xaa6 Cys (SEQ ID NO: 4) where 
 Xaa1 is selected from Trp, DTrp, Tyr, Phe, hPhe, Ala, MeY, Arg, Ben, Nap, Orn, pGlu, DpGlu and a deletion; 
 Xaa2 is selected from Arg, Ala, Asn, Lys, Phe, BHK, Orn, Lys, DArg, Nle, DLys, DMK, DAsn, Thr, ABZ, Nap, Cit, Val, Tyr, Trp, pGlu, DpGlu or a deletion; 
 Xaa3 is selected from Gly, Asp, Lys, Arg, Ala, Nle, Ser or Phe; 
 Xaa4 is selected from Val, Leu, Nle, Ile, Thr, Ala, Asn, Trp, Phe and Abu, and 
 Xaa5 and Xaa6 are independently absent or represent any amino acid residue except Cys; 
 
       or a sequence where one or more of the loop 1 residues Gly, Tyr, Lys and Leu are subject to conservative amino acid substitution or side chain modification; 
       and or a salt, ester, amide, prodrug or cyclised derivative thereof. 
     
     
         3 . The χ-conotoxin peptide according to  claim 2  comprising the following sequence of amino acids:
 Xaa1 Xaa2 Xaa3 Xaa4 Cys Cys Gly Tyr Lys Leu Cys Xaa5 Xaa6 Cys (SEQ ID NO: 4) where Xaa1 is selected from Trp, Tyr, Phe, hPhe, Ala, MeY, Arg, Ben and Nap, 
 Xaa2 is selected from Arg, Asn, Lys, BHK, Orn, Lys, DArg, Nle, DLys, DMK, DAsn, Thr, ABZ, Nap, Cit and Val, 
 Xaa3 is selected from Gly, Asp, Lys, Arg, Ala, Nle and Ser, 
 Xaa4 is selected from Val, Leu, Nle, Ile, Thr, Ala and Abu, and 
 Xaa5 and Xaa6 are independently absent or represent any amino acid residue except Cys; 
 
       or such a sequence where one or more of the loop 1 residues Gly, Tyr, Lys and Leu are subject to conservative amino acid substitution or side chain modification, 
       or a salt, ester, amide, prodrug or cyclised derivative thereof. 
     
     
         4 . The χ-conotoxin peptide according to  claim 3  consisting of the following sequence of amino acids:
 Xaa1 Xaa2 Xaa3 Xaa4 Cys Cys Gly Tyr Lys Leu Cys Xaa5 Xaa6 Cys (SEQ ID NO: 4) where Xaa1 is selected from Trp, Tyr, Phe, hPhe, Ala, MeY, Arg, Ben and Nap, 
 Xaa2 is selected from Arg, Asn, Lys, BHK, Orn, Lys, DArg, Nle, DLys, DMK, DAsn, Thr, ABZ, Nap, Cit and Val, 
 Xaa3 is selected from Gly, Asp, Lys, Arg, Ala, Nle and Ser, 
 Xaa4 is selected from Val, Leu, Nle, Ile, Thr, Ala and Abu, and 
 Xaa5 and Xaa6 are independently absent or represent any amino acid residue except Cys, 
 
       or such a sequence where one or more of the loop 1 residues Gly, Tyr, Lys and Leu are subject to conservative amino acid substitution or side chain modification or a salt, ester, amide or prodrug thereof. 
     
     
         5 . The χ-conotoxin peptide according to  claim 1  comprising the following sequence of amino acids: 
       
         
           
                 
                 
               
                   SEQ ID NO. 5 
                     
                 
                 
                 
               
                   Xaa1 Xaa2 Xaa3 Xaa4 Cys Cys Gly Tyr Lys Leu Cys 
                     
                 
                     
                 
                   Xaa5 Xaa6 Cys 
                 
             
                
               
            
             
                
                
                
               
            
           
         
       
       where
 Xaa1 is an N-terminal residue and is selected from pGlu, DpGlu, Pro, Hyp or an N-acetylated amino acid residue; 
 Xaa2 is selected from Arg, Asn, Lys, BHK, Orn, Lys, DArg, Nle, DLys, DMK, DAsn, Thr, ABZ, Nap, Cit, Val and a deletion, 
 Xaa3 is selected from Gly, Asp, Lys, Arg, Ala, Nle and Ser, 
 Xaa4 is selected from Val, Leu, Nle, Ile, Thr, Ala and Abu, and 
 Xaa5 and Xaa6 are independently absent or represent any amino acid residue except Cys; 
 
       or such a sequence where one or more of the loop 1 residues Gly, Tyr, Lys and Leu are subject to conservative amino substitution or sidechain modification, or a salt, ester, amide or prodrug thereof. 
     
     
         6 . The χ-conotoxin peptide according to  claim 5  consisting of the following sequence of amino acids: 
       
         
           
                 
                 
               
                   SEQ ID NO. 5 
                     
                 
                 
                 
               
                   Xaa1 Xaa2 Xaa3 Xaa4 Cys Cys Gly Tyr Lys Leu Cys 
                     
                 
                     
                 
                   Xaa5 Xaa6 Cys 
                 
             
                
               
            
             
                
                
                
               
            
           
         
       
       where
 Xaa1 is an N-terminal residue and is selected from pGlu, Pro, Hyp or an N-acetylated amino acid residue; 
 Xaa2 is selected from Arg, Asn, Lys, BHK, Orn, Lys, DArg, Nle, DLys, DMK, DAsn, Thr, ABZ, Nap, Cit, pGlu, Val and a deletion, 
 Xaa3 is selected from Gly, Asp, Lys, Arg, Ala, Nle and Ser, 
 Xaa4 is selected from Val, Leu, Nle, Ile, Thr, Ala and Abu, and 
 Xaa5 and Xaa6 are independently absent or represent any amino acid residue except Cys; 
 
       or such a sequence where one or more of the loop 1 residues Gly, Tyr, Lys and Leu are subject to conservative amino and substitution or said chain modification, or a salt or prodrug thereof. 
     
     
         7 . The χ-conotoxin peptide according to  claim 2  comprising the following sequence of amino acids: 
       
         
           
                 
                 
               
                   SEQ ID NO. 6 
                     
                 
                 
                 
               
                   Xaa2 Xaa3 Xaa4 Cys Cys Gly Tyr Lys Leu Cys Xaa5 
                     
                 
                     
                 
                   Xaa6 Cys 
                 
             
                
               
            
             
                
                
                
               
            
           
         
       
       where
 Xaa2 is BHK, Orn, Arg, DArg or DMK; 
 Xaa3 is selected from Gly, Asp, Lys, Arg, Ala, Nle and Ser, 
 Xaa4 is selected from Val, Leu, Nle, Ile, Thr, Ala and Abu, and 
 Xaa5 and Xaa6 are independently absent or represent any amino acid residue except Cys; 
 
       or such a sequence where one or more of the loop 1 residues Gly, Tyr, Lys and Leu are subject to conservative amino acid or side chain modification, or a salt, ester, amide, prodrug or cyclised derivative thereof. 
     
     
         8 . The χ-conotoxin peptide according to  claim 7  consisting of the following sequence of amino acids: 
       
         
           
                 
                 
               
                   SEQ ID NO. 6 
                     
                 
                 
                 
               
                   Xaa2 Xaa3 Xaa4 Cys Cys Gly Tyr Lys Leu Cys Xaa5 
                     
                 
                     
                 
                   Xaa6 Cys 
                 
             
                
               
            
             
                
                
                
               
            
           
         
       
       where
 Xaa2 is BHK, Orn, Arg, DArg or DMK; 
 Xaa3 is selected from Gly, Asp, Lys, Arg, Ala, Nle and Ser; 
 Xaa4 is selected from Val, Leu, Nle, Ile, Thr, Ala and Abu; and 
 Xaa5 and Xaa6 are independently absent or represent any amino acid residue except Cys; 
 
       or such a sequence where one or more of the loop 1 residues Gly, Tyr, Lys and Leu are subject to conservative amino acid or side chain modification, or a salt, ester, amide, prodrug or cyclised derivative thereof. 
     
     
         9 . The peptide according to  claim 2  wherein Xaa1 is Trp, Tyr or hPhe. 
     
     
         10 . The peptide according to  claim 9  wherein Xaa1 is Trp. 
     
     
         11 . The peptide according to  claim 2  wherein Xaa2 is Arg, Lys or Asn. 
     
     
         12 . The peptide according to  claim 5  wherein Xaa1 is pGlu or DpGlu. 
     
     
         13 . The peptide according to  claim 5  wherein Xaa2 is a deletion. 
     
     
         14 . The peptide according to  claim 5  wherein Xaa2 is BHK or Orn. 
     
     
         15 . The peptide according to  claim 2  wherein Xaa3 is Gly or Asp. 
     
     
         16 . The peptide according to  claim 15  wherein Xaa3 is Gly. 
     
     
         17 . The peptide according to  claim 2  wherein Xaa4 is Leu, Nle or Val. 
     
     
         18 . The peptide according to  claim 2  wherein Xaa5 is selected from the group consisting of His, Arg, Trp, NaI, Glu and a deletion. 
     
     
         19 . The peptide according to  claim 18  wherein Xaa5 is Arg or His. 
     
     
         20 . The peptide according to  claim 2  wherein Xaa6 is selected from the group consisting of Hyp, Pro, Ala, Tic, Pip, MeY, DMD, Phe, THZ, Glu, Nle, Tyr and a deletion. 
     
     
         21 . The peptide according to  claim 20  wherein Xaa6 is Hyp or Pro. 
     
     
         22 . The peptide according to  claim 1  wherein the Tyr of loop 1 has been replaced with MeY and/or the Leu of loop 1 is replaced with Hle or Nle. 
     
     
         23 . The peptide according to  claim 5  wherein the Tyr of loop 1 has been replaced with MeY and/or the Leu of loop 1 is replaced with Hle or Nle. 
     
     
         24 . The peptide according to  claim 1  having from 11 to 20 amino acids. 
     
     
         25 . An isolated, synthetic or recombinant χ-conotoxin peptide as set forth in Table 2. 
     
     
         26 . An isolated, synthetic or recombinant peptide as set forth in Table 3, excluding SEQ ID NO. 1 and 7. 
     
     
         27 . The peptide according to  claim 1  with the ability to selectively inhibit neuronal noradrenaline transporter, and has negligible or no substantial anticholinergic effect. 
     
     
         28 . A composition comprising an isolated, synthetic or recombinant χ-conotoxin peptide having the ability to inhibit neuronal noradrenaline transporter, wherein said χ-conotoxin peptide comprises the following sequence of amino acids: 
       
         
           
                 
                 
               
                   (SEQ ID NO: 3) 
                     
                 
                 
                 
                 
               
                     
                   Cys Cys Gly Tyr Lys Leu Cys Xaa5 Xaa6 Cys 
                     
                 
             
                
               
            
             
                
               
            
           
         
       
       where Xaa5 and Xaa6 are independently absent or represent any amino acid residue except Cys, or such a sequence in which loop 1 residues Gly, Tyr, Lys or Leu are subject to conservative amino acid substitution or side chain modification, with the proviso that the peptide is not χ-MrIA or χ-MrIB; or a salt, ester, amide, prodrug or cyclised derivative thereof, 
       and a pharmaceutically acceptable carrier or diluent. 
     
     
         29 . The composition of  claim 28  wherein the peptide is as defined in any one of  claims 2 - 8 . 
     
     
         30 . A method for the treatment or prophylaxis of urinary or cardiovascular conditions or diseases or mood disorders or for the treatment or control of pain or inflammation including the step of administering to a mammal an effective amount of an isolated, synthetic or recombinant χ-conotoxin peptide having the ability to inhibit neuronal noradrenaline transporter, wherein said χ-conotoxin peptide comprises the following sequence of amino acids: 
       
         
           
                 
                 
               
                   (SEQ ID NO: 3) 
                     
                 
                 
                 
                 
               
                     
                   Cys Cys Gly Tyr Lys Leu Cys Xaa5 Xaa6 Cys 
                     
                 
             
                
               
            
             
                
               
            
           
         
       
       where Xaa5 and Xaa6 are independently absent or represent any amino acid residue except Cys, or such a sequence in which Gly, Tyr, Lys or Leu are subject to conservative amino acid substitution or side chain modification, with the proviso that the peptide is not χ-MrIA or χ-MrIB; or a salt, ester, amide, prodrug or cyclised derivative thereof. 
     
     
         31 . The method of  claim 30  wherein the peptide is as defined in any one of  claims 2 - 8 . 
     
     
         32 . The method according to  claim 30  wherein the peptide is administered substantially simultaneously or sequentially with other active agents useful in the treatment of the conditions, diseases or disorders.