Treatment of pain
View Patent ↗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 can cleave a protein of the exocytic fusion apparatus of a nociceptive sensory afferent; a Targeting Moiety that can bind to a Binding Site on the nociceptive sensory afferent, which Binding Site can undergo 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 translation domain that can translocate the protease or protease fragment from within an endosome, across the endosomal membrane and into the cytosol of the nociceptive sensory afferent.
1. A method of preventing or suppressing a specific pain condition in a subject, wherein said pain condition is selected from the group consisting of: neuropathic pain, inflammatory pain, headache pain, somatic pain, visceral pain and referred pain, said method comprising administering to said subject a therapeutically effective amount of a polypeptide fusion protein, comprising:
a) a clostridial neurotoxin protease or an IgA protease that cleaves a protein of the exocytic fusion apparatus in a nociceptive sensory afferent;
b) a Targeting Moiety that binds to a Binding Site present on the nociceptive sensory afferent;
c) a protease cleavage site located between the clostridial neurotoxin protease or the IgA protease and the Targeting Moiety; and
d) a translocation domain that translocates the protease from within an endosome, across an endosomal membrane and into the cytosol of the nociceptive sensory afferent;
wherein the Targeting Moiety is located between the protease cleavage site and the translocation domain and wherein,
I. the clostridial neurotoxin protease or the IgA protease forms a disulphide bond with the translocation domain and,
wherein the amino acid residue of the clostridial neurotoxin protease or the IgA protease that forms the first part of the disulphide bond is located within the last 20 C-terminal amino acid residues of the clostridial neurotoxin protease or the IgA protease, and the amino acid residue within the translocation domain that forms the second part of the disulphide bond is located within the first 20 N-terminal amino acid residues of the translocation domain; or
II. the clostridial neurotoxin protease or the IgA protease forms a disulphide bond with the Targeting Moiety component of the polypeptide fusion protein,
wherein the amino acid residue of the clostridial neurotoxin protease or the IgA protease that forms the first part of the disulphide bond is located within the last 20 C-terminal amino acid residues of the clostridial neurotoxin protease or the IgA protease, and the amino acid residue within the Targeting Moiety that forms the second part of the disulphide bond is located within the last 20 C-terminal amino acid residues of the Targeting Moiety.
2. The method according to claim 1 , wherein the Targeting Moiety and the protease cleavage site are separated by at most 10 amino acid residues.
3. The method according to claim 1 , wherein the Targeting Moiety and the protease cleavage site are separated by at most 5 amino acid residues, or by at most 0 amino acid residues.
4. The method according to claim 1 , wherein the translocation domain comprises a clostridial neurotoxin peptide.
5. The method according to claim 1 , wherein the Targeting Moiety binds to an ORL1 receptor.
6. The method according to claim 1 , wherein the Targeting Moiety has at least 80% sequence identity to SEQ ID No: 38.
7. The method according to claim 1 , wherein the Targeting Moiety has at least 80% sequence identity to SEQ ID No: 40.
8. The method according to claim 1 , wherein the Targeting Moiety has at least 80% sequence identity to SEQ ID No: 42.
9. The method according to claim 1 , wherein the Targeting Moiety has at least 80% sequence identity to SEQ ID No: 44.
10. The method according to claim 1 , wherein the Targeting Moiety has at least 80% sequence identity to SEQ ID No: 46.
11. The method according to claim 1 , wherein the Targeting Moiety has at least 80% sequence identity to SEQ ID No: 48.
12. The method according to claim 1 , wherein the Targeting Moiety has at least 80% sequence identity to SEQ ID No: 50.
13. The method according to claim 1 , wherein the Targeting Moiety is selected from the group consisting of: opioid, nociceptin; β-endorphin, endomorphine-1, endomorphine-2, dynorphin, met-enkephalin, leu-enkephalin, galanin, and PAR-2 peptide.
14. The method according to claim 1 , wherein the polypeptide fusion protein comprises a polypeptide sequence having at least 80% sequence identity to a polypeptide sequence according to a SEQ ID NO: selected from the group consisting 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, and 88.
15. A method according to claim 1 , wherein the polypeptide fusion protein is in the form of a di-chain polypeptide that has been prepared by a process comprising:
a) contacting a single-chain polypeptide fusion protein with a protease capable of cleaving the single-chain polypeptide fusion protein; and
b) cleaving the single-chain polypeptide fusion protein at the protease cleavage site; and thereby forming the di-chain polypeptide.
16. The method according to claim 15 , wherein the protease cleavage site is cleaved by a protease selected from the group consisting of enterokinase, Factor X, TEV (Tobacco Etch virus), Thrombin, and PreScission.
17. The method according to claim 16 , wherein the protease cleavage site is integrated at a position within the fusion protein such that cleavage of the integrated protease converts the single-chain fusion protein into the di-chain polypeptide in which the Targeting Moiety has a free amino terminus that interacts directly with the Binding Site.
18. The method according to claim 1 , wherein the inflammatory pain is selected from a pain associated with an inflammatory condition selected from the group consisting of arthritic disorder; autoimmune disease; connective tissue disorder; injury; infection; neuritis; and joint inflammation.
19. The method according to claim 1 , wherein the headache pain is selected from the group consisting of muscular/myogenic headache; vascular headache; high blood pressure headache; fraction and inflammatory headache; hormone headache; rebound headache; chronic sinusitis headache; organic headache; and ictal headache.
20. The method according to claim 1 , wherein the somatic pain is selected from the group consisting of excessive muscle tension; repetitive motion disorder; muscle disorder; myalgia; infection; and drugs.
21. The method according to claim 1 , wherein the visceral pain is selected from the group consisting of functional visceral pain; chronic gastrointestinal inflammation; autoimmune pain; organic visceral pain; and treatment-induced visceral pain.
22. The method according to claim 1 , wherein the referred pain is selected from the group consisting of pain associated with intervertebral disc herniation and myocardial ischemia.