COMPOSITIONS AND METHODS FOR INHIBITING SHIGA TOXIN AND SHIGA-LIKE TOXIN

• US 20090280104A1
• Filed: 05/11/2009
• Published: 11/12/2009
• Est. Priority Date: 05/09/2008
• Status: Abandoned Application

• Current Assignee: Recopharma AB
• Original Assignee: Recopharma AB
• Inventors: Gustafsson, Anki, Holgersson, Jan
• Application Number:
  US12/463,943
• Publication Number:
  US 20090280104A1
• Time in Patent Office: Days
• US Class Current: 424/94.500
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: C07K 14/4727 Mucins, e.g. human intestinal mucin
• CPC Class Codes: C07K 14/70589 CD45
• CPC Class Codes: C07K 14/70596 Molecules with a "CD"-designation not provided for elsewhere
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 2319/32 fusions with soluble part of a cell surface receptor, "decoy receptors"
• CPC Class Codes: C07K 2319/91 containing a motif for glycosylation

Abstract

The present invention provides compositions and methods for treating or preventing infection by shiga toxin producing bacteria.

Claims

1. A fusion polypeptide comprising a first polypeptide linked to a second polypeptide, wherein the first polypeptide is a mucin polypeptide glycosylated by an α

1,4galactosyltransferase, and the second polypeptide comprises at least a region of an immunoglobulin polypeptide.

2. The fusion polypeptide of claim 1, wherein said mucin polypeptide is further glycosylated by a core 2β

1,6-N-acetylglucosaminyltransferase.

3. The fusion polypeptide of claim 1 or 2, wherein said mucin polypeptide has a glycan repertoire including a Galα

4Galβ

3GalNacα

structure or a Galα

4Galβ

4GlcNac structure.

4. The fusion polypetide of claim 1, wherein said mucin polypeptide is selected from the group consisting of PSGL-1, MUC1, MUC2, MUC3, MUC4, MUC5a, MUC5b, MUC5c, MUC6, MUC11, MUC12, CD34, CD43, CD45, CD96, GlyCAM-1, and MAdCAM-1 or fragment thereof.

5. The fusion polypeptide of claim 4, wherein said mucin polypeptide comprises at least a region of a P-selectin glycoprotein ligand-1 (PSGL-1).

6. The fusion polypeptide of claim 5, wherein said mucin polypeptide includes an extracellular portion of a P-selectin glycoprotein ligand-1.

7. The fusion polypeptide of claim 1, wherein the second polypeptide comprises a region of a heavy chain immunoglobulin polypeptide.

8. The fusion polypeptide of claim 1, wherein said second polypeptide comprises an Fc region of an immunoglobulin heavy chain.

9. A method for preventing or alleviating a symptom of bacterial toxin infection in a subject in need thereof, the method comprising administering to the subject fusion polypeptide of claim 1.

10. The method of claim 9, wherein said fusion polypeptide is administered to the subject systemically.

11. The method of claim 9, wherein said fusion polypeptide is administered to the subject rectally.

12. The method of claim 9, wherein said bacterial toxin is produced by a bacteria selected from the group consisting of Shigella dysenteria, enterohaemorrhagic E. coli, Aeromononas caviae, Aeromononas hydrophila, Citrobacter freundii, and Enterobacter cloacae.

13. The method of claim 12, wherein the bacterial toxin is Shiga toxin or Shiga-like toxin 1.

14. The method of claim 12, wherein the bacterial toxin is Shiga-like toxin 2.

15. A method of producing a mucin-immunoglobulin fusion polypeptide comprising:

a) providing a cell comprising;

i) a nucleic acid encoding a mucin polypeptide linked to a nucleic acid encoding at least a portion of an immunoglobulin polypeptide;

ii) a nucleic acid encoding a α

1,4galactosyltransferase polypeptide; and

iii) optionally a nucleic acid encoding a core 2β

1,6-N-acetylglucosaminyltransferase; and

b) culturing the cell under conditions that permit production of said mucin-immunoglobulin fusion polypeptide wherein said fusion polypeptide has a glycan repertoire including a Galα

4Galβ

3GalNAc α

structure or a Galα

4Galβ

4GlcNac structure; and

c) isolating said mucin-immunoglobulin fusion polypeptide.

16. The method of claim 15, wherein said cell is a eukaryotic cell or a prokaryotic cell.

17. The method of claim 16, wherein said eukaryotic cell is a mammalian cell, or a yeast cell.

18. The method of claim 17, wherein said mammalian cell is a CHO cell.

19. The method of claim 16, wherein said prokaryotic cell is a bacterial cell.

20. A cell comprising:

a) a nucleic acid encoding a mucin polypeptide linked to a nucleic acid encoding at least a portion of an immunoglobulin polypeptide;

b) a nucleic acid encoding a α

1,4galactosyltransferase polypeptide; and

c) optionally a nucleic acid encoding a core 2β

1,6-N-acetylglucosaminyltransferase.