Fusion proteins comprising a non-cytotoxic protease, a translocation domain, and a targeting moiety that binds a galanin receptor and methods for treating, preventing or ameliorating pain using such fusion proteins

Patent metadata

Field	Value
Publication number	US-11248219-B2
Application number	US-201916416435-A
Country	US
Kind code	B2
Filing date	May 20, 2019
Priority date	Aug 27, 2012
Publication date	Feb 15, 2022
Grant date	Feb 15, 2022

Abstract

Single chain polypeptide fusion protein, comprising: a non-cytotoxic protease capable of cleaving a protein of the exocytic fusion apparatus of a nociceptive sensory afferent; a galanin targeting moiety; a protease cleavage site; a translocation domain; a first spacer located between the non-cytotoxic protease and the protease cleavage site; and a second spacer located between the galanin targeting moiety and the translocation domain.

First claim

The invention claimed is:

1. A fusion protein comprising an analgesic, non-cytotoxic single chain polypeptide, the polypeptide comprising:
a non-cytotoxic protease capable of cleaving a protein of the exocytic fusion apparatus of a nociceptive sensory afferent;
a targeting moiety that binds to a binding site on the nociceptive sensory afferent, wherein the binding site is capable of being incorporated into an endosome within the nociceptive sensory afferent;
a protease cleavage site;
a translocation domain capable of translocating the protease from within the endosome, across the endosomal membrane, and into the cytosol of the nociceptive sensory afferent;
a first spacer located between the non-cytotoxic protease and the protease cleavage site, the first spacer comprising an amino acid sequence of from 4 to 25 amino acid residues; and
a second spacer located between the targeting moiety and the translocation domain, the second spacer comprising an amino acid sequence of from 4 to 35 amino acid residues;

wherein:
the protease cleavage site is located between the non-cytotoxic protease and the targeting moiety;
the targeting moiety is located between the protease cleavage site and the translocation domain; and
the analgesic, single-chain polypeptide has at least 99% sequence identity to SEQ ID NO: 32.

2. The fusion protein of claim 1 , wherein the first spacer consists of an amino acid sequence of from 6 to 16 amino acid residues.

3. The fusion protein of claim 1 , wherein the first spacer comprises amino acid residues selected from the group consisting of: glycine, threonine, arginine, serine, alanine, asparagine, glutamine, aspartic acid, proline, glutamic acid, and lysine.

4. The fusion protein of claim 1 , wherein the first spacer comprises amino acid residues selected from the group consisting of: glycine, serine, and alanine.

5. The fusion protein of claim 1 , wherein the first spacer is a GS5 spacer.

6. The fusion protein of claim 1 , wherein the targeting moiety comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 8.

7. The fusion protein of claim 1 consisting of an analgesic, single-chain polypeptide having at least 99% sequence identity with the sequence of SEQ ID NO: 32.

8. The fusion protein of claim 1 comprising the analgesic, single-chain polypeptide of SEQ ID NO: 32.

9. The fusion protein of claim 1 consisting of the analgesic, single-chain polypeptide of SEQ ID NO: 32.

Assignees

• Ipsen Bioinnovation Ltd
• Allergan Inc

Inventors

• James Peter
• Foster Keith
• Chaddock John
• Aoki Roger
• Steward Lance
• Francis Joseph

Classifications

• C12N15/62

  DNA sequences coding for fusion proteins · CPC title

• C07K19/00

  Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes} · CPC title

• C07K14/575

  Hormones (derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695) · CPC title

• A61K38/16

  Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof {(enzyme inhibitors A61K38/005)} · CPC title

• C12Y304/24069

  Bontoxilysin (3.4.24.69), i.e. botulinum neurotoxin · CPC title