Polymer-conjugated glycosylated neublastin

• US 8,642,732 B2
• Filed: 03/21/2012
• Issued: 02/04/2014
• Est. Priority Date: 04/18/2003
• Status: Active Grant

• Current Assignee: Sinfonia Biotherapeutics AB
• Original Assignee: Biogen Idec MA Inc. (Biogen, Inc.)
• Inventors: Sah, Dinah Wen-Yee, Pepinsky, R. Blake, Rossomando, Anthony
• Primary Examiner(s): Gamett, Daniel C
• Application Number:
  US13/425,583
• Publication Number:
  US 20120316109A1
• Time in Patent Office: 685 Days
• Field of Search: None
• US Class Current: 530/350
• 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 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 25/04 Centrally acting analgesics, e.g. opioids
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4756 Neuregulins, i.e. p185erbB2 ligands, glial growth factor, heregulin, ARIA, neu differentiation factor

Abstract

The following class of molecule is disclosed: a dimer containing a first neublastin polypeptide and a second neublastin polypeptide, wherein: (a) at least one of the polypeptides is glycosylated; (b) at least one of the polypeptides is conjugated at its N-terminus to a water-soluble synthetic polymer; and (c) neither of the polypeptides is conjugated to a water-soluble synthetic polymer at a position other than the N-terminus. Such dimers possess the biological activity of wild-type neublastin while displaying enhanced serum half-life and enhanced potency relative to wild-type neublastin.

Claims

1. A dimer comprising a first neublastin polypeptide and a second neublastin polypeptide, wherein:

(a) at least one of the polypeptides is glycosylated;

(b) at least one of the polypeptides is conjugated at its N-terminus to a water-soluble synthetic polymer; and

(c) neither of the polypeptides is conjugated to a water-soluble synthetic polymer at a position other than the N-terminus, and wherein the amino acid sequence of the first neublastin polypeptide and the second neublastin polypeptide are the same and are selected from the group consisting of NBN113 (SEQ ID NO;

2), NBN140 (SEQ ID NO;

6), NBN116 (SEQ ID NO;

7), NBN112 (SEQ ID NO;

8), NBN111 (SEQ ID NO;

9), NBN110 (SEQ ID NO;

10), NBN109 (SEQ ID NO;

11), NBN108 (SEQ ID NO;

12), NBN107 (SEQ ID NO;

13), NBN106 (SEQ ID NO;

14), NBN105 (SEQ ID NO;

15), NBN104 (SEQ ID NO;

16), NBN103 (SEQ ID NO;

17), NBN102 (SEQ ID NO;

18), NBN101 (SEQ ID NO;

19), NBN100 (SEQ ID NO;

20) and NBN99 (SEQ ID NO;

21).

2. The dimer of claim 1, wherein the water-soluble synthetic polymer is a polyalkylene glycol moiety.

3. The dimer of claim 1, wherein the N-terminal amino acid of the first neublastin polypeptide and the N-terminal amino acid of the second neublastin polypeptide each is conjugated to a polyalkylene glycol moiety.

4. The dimer of claim 2, wherein the polyalkylene glycol moiety is a polyethylene glycol moiety.

5. The dimer of claim 4, wherein at least one of the polypeptides is glycosylated at the asparagine residue corresponding to the asparagine at position 95 of SEQ ID NO:

2.

6. The dimer of claim 1, wherein the N-terminal amino acid of the first neublastin polypeptide and the N-terminal amino acid of the second neublastin polypeptide each is conjugated to a polyethylene glycol moiety.

7. The dimer of claim 6, wherein at least one of the polypeptides is glycosylated at the asparagine residue corresponding to the asparagine at position 95 of SEQ ID NO:

2.

8. A composition comprising the dimer of claim 1 and a pharmaceutically acceptable carrier.

9. A method of treating neuropathic pain in a human in need thereof, comprising administering to the human a therapeutically effective amount of the dimer of claim 1.

10. A method of treating a peripheral neuropathy in a human in need thereof, comprising administering to the human a therapeutically effective amount of the dimer of claim 1.

11. A method of treating painful diabetic neuropathy in a human in need thereof, comprising administering to the human a therapeutically effective amount of the dimer of claim 1.

12. A dimer comprising a first neublastin polypeptide and a second neublastin polypeptide, wherein:

(a) at least one of the polypeptides is glycosylated;

(b) at least one of the polypeptides is conjugated at its N-terminus to a polyalkylene glycol moiety; and

(c) neither of the polypeptides is conjugated to a polyalkylene glycol moiety at a position other than the N-terminus, and wherein the amino acid sequence of the first neublastin polypeptide and the second neublastin polypeptide is NBN104 (SEQ ID NO;

16).

13. The dimer of claim 12, wherein the N-terminal amino acid of the first neublastin polypeptide and the N-terminal amino acid of the second neublastin polypeptide each is conjugated to a polyalkylene glycol moiety.

14. A composition comprising the dimer of claim 12 and a pharmaceutically acceptable carrier.

15. A method of treating neuropathic pain in a human in need thereof, comprising administering to the human a therapeutically effective amount of the dimer of claim 12.

16. A method of treating a peripheral neuropathy in a human in need thereof, comprising administering to the human a therapeutically effective amount of the dimer of claim 12.

17. A method of treating painful diabetic neuropathy in a human in need thereof, comprising administering to the human a therapeutically effective amount of the dimer of claim 12.

18. A homodimer comprising a first neublastin polypeptide and a second neublastin polypeptide, wherein the amino acid sequence of the first neublastin polypeptide and the second neublastin polypeptide is NBN104 (SEQ ID NO:

16), wherein each of the neublastin polypeptides is conjugated to a polyethylene glycol moiety at its amino terminus and is glycosylated at the asparagine residue corresponding to the asparagine at position 95 of SEQ ID NO;

2, and wherein neither of the polypeptides is conjugated to a polyethylene glycol moiety at a position other than the N-terminus.

19. A composition comprising the homodimer of claim 18 and a pharmaceutically acceptable carrier.

20. A method of treating neuropathic pain in a human in need thereof, comprising administering to the human a therapeutically effective amount of the homodimer of claim 18.

21. A method of treating a peripheral neuropathy in a human in need thereof, comprising administering to the human a therapeutically effective amount of the homodimer of claim 18.

22. A method of treating painful diabetic neuropathy in a human in need thereof, comprising administering to the human a therapeutically effective amount of the homodimer of claim 18.