Treatment of pain

Abstract

Use of a therapeutic molecule, for the treatment of specific pain conditions, wherein the therapeutic molecule is a single chain, polypeptide fusion protein, comprising: a non-cytotoxic protease, or a fragment thereof, which protease or protease fragment is capable of cleaving a protein of the exocytic fusion apparatus of a nociceptive sensory afferent; a Targeting Moiety that is capable of binding to a Binding Site on the nociceptive sensory afferent, which Binding Site is capable of undergoing endocytosis to be incorporated into an endosome within the nociceptive sensory afferent; a protease cleavage site at which site the fusion protein is cleavable by a protease, wherein the protease cleavage site is located between the non-cytotoxic protease or fragment thereof and the Targeting Moiety; and a translocation domain that is capable of translocating the protease or protease fragment from within an endosome, across the endosomal membrane and into the cytosol of the nociceptive sensory afferent.

1 . Use of a therapeutic molecule for the manufacture of a medicament for the treatment of particular types of pain, wherein the therapeutic molecule is a single chain, polypeptide fusion protein, comprising:
 a. a non-cytotoxic protease, or a fragment thereof, which protease or protease fragment is capable of cleaving a protein of the exocytic fusion apparatus of a nociceptive sensory afferent;   b. a Targeting Moiety that is capable of binding to a Binding Site on the nociceptive sensory afferent, which Binding Site is capable of undergoing endocytosis to be incorporated into an endosome within the nociceptive sensory afferent;   c. a protease cleavage site at which site the fusion protein is cleavable by a protease, wherein the protease cleavage site is located between the non-cytotoxic protease or fragment thereof and the Targeting Moiety; and   d. a translocation domain that is capable of translocating the protease or protease fragment from within an endosome, across the endosomal membrane and into the cytosol of the nociceptive sensory afferent.   
     
     
         2 . Use according to  claim 1 , wherein the Targeting Moiety and the protease cleavage site are separated by at most 10 amino acid residues, preferably by at most 5 amino acid residues, and more preferably by at most zero amino acid residues. 
     
     
         3 . Use according to  claim 1  or  claim 2 , wherein the Targeting Moiety is located between the protease cleavage site and the translocation domain. 
     
     
         4 . Use according to any preceding claim, wherein the non-cytotoxic protease is a clostridial neurotoxin L-chain or an IgA protease. 
     
     
         5 . Use according to any preceding claim, wherein the translocation domain is the H N  domain of a clostridial neurotoxin. 
     
     
         6 . Use according to any preceding claim, wherein the Targeting Moiety comprises at most 50 amino acid residues, preferably at most 40 amino acid residues, more preferably at least 30 amino acid residues, and most preferably at most 20 amino acid residues. 
     
     
         7 . Use according to any of  claims 1 - 6 , wherein the Targeting Moiety is an opioid. 
     
     
         8 . Use according to any of  claim 1 - 6 , wherein the Targeting Moiety is an agonist of a receptor present on a nociceptive sensory afferent. 
     
     
         9 . Use according to  claim 8 , wherein the Targeting Moiety is an agonist of a receptor present on a primary nociceptive sensory afferent. 
     
     
         10 . Use according to any of  claims 1 - 6 , wherein the Targeting Moiety binds to the ORL 1  receptor. 
     
     
         11 . Use according to  claim 10 , wherein the Targeting Moiety binds specifically to the ORL 1  receptor. 
     
     
         12 . Use according to  claim 10  or  11 , wherein the Targeting Moiety is an agonist of the ORL 1  receptor. 
     
     
         13 . Use according to any one of  claims 10 - 12 , wherein the Targeting Moiety has at least 70% homology to SEQ ID No. 38 or a fragment thereof. 
     
     
         14 . Use according to  claim 13 , wherein the Targeting Moiety as at least 80% homology to SEQ ID No. 38 or a fragment thereof. 
     
     
         15 . Use according to  claim 14 , wherein the Targeting Moiety has at least 90% homology to SEQ ID No. 38 or a fragment thereof. 
     
     
         16 . Use according to  claim 15 , wherein the Targeting Moiety has at least 95% homology to SEQ ID No. 38 or a fragment thereof. 
     
     
         17 . Use according to any one of  claims 10 - 12 , wherein the Targeting Moiety is SEQ ID No. 38 or a fragment thereof. 
     
     
         18 . Use according to any of  claims 10 - 12 , wherein the Targeting Moiety is one of SEQ ID Nos: 40, 42, 44, 46, 48 or 50. 
     
     
         19 . Use according to any one of  claims 10 - 12 , wherein the Targeting Moiety is nociceptin. 
     
     
         20 . Use according to any of  claims 1 - 6 , wherein the Targeting Moiety is selected from the group consisting of nociceptin, β-endorphin, endomorphine-1, endomorphine-2, dynorphin, met-enkephalin, leu-enkephalin, galanin, and PAR-2 peptide. 
     
     
         21 . Use according to any preceding claim, wherein the fusion protein comprises a purification tag. 
     
     
         22 . Use according to  claim 21 , wherein the fusion protein comprises a purification tag, which is present at the N-terminal and/or C-terminal end of the fusion protein. 
     
     
         23 . Use according to  claim 21  or  22 , wherein the purification tag is joined to the fusion protein by a peptide spacer molecule. 
     
     
         24 . Use according to any preceding claim, wherein the translocation domain is separated from the Targeting Moiety by a peptide spacer molecule. 
     
     
         25 . Use according to any preceding claim, wherein the polypeptide fusion protein comprising any one of SEQ ID NOs: 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 52, 59, 61, 64, 67, 69, 71, 73, 76, 79, 82, 85, or 88. 
     
     
         26 . A method of preparing a non-cytotoxic fusion protein, comprising:
 a. contacting a single-chain polypeptide fusion protein as defined in any of  claims 1 - 25  with a protease capable of cleaving the protease cleavage site;   b. cleaving the protease cleavage site; and thereby forming a di-chain fusion protein.   
     
     
         27 . A non-cytotoxic fusion protein, obtainable by the method of  claim 26 , wherein the protein is a di-chain polypeptide, and wherein:
 a. the first chain comprises the non-cytotoxic protease, or a fragment thereof, which protease or protease fragment is capable of cleaving a protein of the exocytic fusion apparatus of a nociceptive sensory afferent;   b. the second chain comprises the TM and the translocation domain that is capable of translocating the protease or protease fragment from within an endosome, across the endosomal membrane and into the cytosol of the nociceptive sensory afferent; and   the first and second chains are disulphide linked together.   
     
     
         28 . Use of a fusion protein according to  claim 27 , for the manufacture of a medicament for treating, preventing or ameliorating particular types of pain. 
     
     
         29 . A method of treating, preventing or ameliorating particular types of pain in a subject, comprising administering to said patient a therapeutically effective amount of a fusion protein as defined in any of  claims 1 - 27 .