Polymer conjugated glycosylated neublastin

• US 8,163,875 B2
• Filed: 04/16/2004
• Issued: 04/24/2012
• 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:
  US10/553,710
• Publication Number:
  US 20070238650A1
• Time in Patent Office: 2,930 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.

2. The dimer according to claim 1, wherein the first neublastin polypeptide is 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).

3. The dimer according to claim 1, wherein the amino acid sequence of the first neublastin polypeptide and the second neublastin polypeptide are the same.

4. The dimer of claim 3, wherein the amino acid sequence of the first neublastin polypeptide is NBN104 (SEQ ID NO:

16).

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

6. The dimer of claim 5, 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.

7. The dimer according to claim 6, wherein the average total molecular weight of the polyalkylene glycol moiety or moieties conjugated to the dimer is 10-50 kDa.

8. The dimer of claim 7, wherein the average total molecular weight of the polyalkylene glycol moiety or moieties conjugated to the dimer is 15-45 kDa.

9. The dimer of claim 8, wherein the average total molecular weight of the polyalkylene glycol moiety or moieties conjugated to the dimer is 20-40 kDa.

10. The dimer according to claim 5, wherein the polyalkylene glycol moiety is linear.

11. The dimer according to claim 5, wherein the polyalkylene glycol is branched.

12. The dimer of claim 5, wherein the polyalkylene glycol moiety is a polyethylene glycol (PEG) moiety.

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

14. A method of treating neuropathic pain in a mammal, comprising administering to the mammal a therapeutically effective amount of the dimer of claim 1.

15. The method of claim 14, wherein the mammal is a human.

16. The method of claim 15, wherein the route of administration is intravenous, intramuscular or subcutaneous.

17. The method claim 14, wherein the therapeutically effective amount is from 0.1 μ

g/kg to 1000 μ

g/kg.

18. The method of claim 17, wherein the therapeutically effective amount is from 1 μ

g/kg to 100 μ

g/kg.

19. The method of claim 18, wherein the therapeutically effective amount is from 1 μ

g/kg to 30 μ

g/kg.

20. The method of claim 19, wherein the therapeutically effective amount is from 3 μ

g/kg to 10 μ

g/kg.

21. A method of treating tactile allodynia in a mammal, comprising administering to the mammal a therapeutically effective amount of the dimer of claim 1.

22. A method of treating thermal hyperalgesia, comprising administering to the mammal a therapeutically effective amount of the dimer of claim 1.

23. A method of activating the RET receptor in a mammal, comprising administering to the mammal an effective amount of the dimer of claim 1.

24. A method of treating neuropathic pain, tactile allodynia or thermal hyperalgesia in a mammal, comprising co-administering to the mammal an effective amount of the dimer of claim 1 and an analgesic agent.

25. 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 polyethylene glycol moiety; and

(c) neither of the polypeptides is conjugated to a polyethylene 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 is NBN104 (SEQ ID NO;

16).

26. A composition comprising the dimer of claim 25 and a pharmaceutically acceptable carrier.

27. A method of treating neuropathic pain in a human, comprising administering to the human a therapeutically effective amount of the dimer of claim 25.

28. A method of activating the RET receptor in a human, comprising administering to the human an effective amount of the dimer of claim 25.

29. A method of treating a peripheral neuropathy in a human, comprising administering to the human a therapeutically effective amount of the dimer of claim 25.

30. A method of treating painful diabetic neuropathy in a human, comprising administering to the human a therapeutically effective amount of the dimer of claim 25.

31. The dimer of claim 25, 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.

32. A composition comprising the dimer of claim 31 and a pharmaceutically acceptable carrier.

33. A method of treating neuropathic pain in a human, comprising administering to the human a therapeutically effective amount of the dimer of claim 31.

34. A method of activating the RET receptor in a human, comprising administering to the human an effective amount of the dimer of claim 31.

35. A method of treating a peripheral neuropathy in a human, comprising administering to the human a therapeutically effective amount of the dimer of claim 31.

36. A method of treating painful diabetic neuropathy in a human, comprising administering to the human a therapeutically effective amount of the dimer of claim 31.

37. 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 polyethylene glycol moiety; and

(c) neither of the polypeptides is conjugated to a polyethylene 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 is NBN113 (SEQ ID NO;

2).

38. The dimer of claim 37, 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.

39. A composition comprising the dimer of claim 37 and a pharmaceutically acceptable carrier.

40. A method of treating neuropathic pain in a human, comprising administering to the human a therapeutically effective amount of the dimer of claim 37.

41. A method of activating the RET receptor in a human, comprising administering to the human an effective amount of the dimer of claim 37.

42. A method of treating a peripheral neuropathy in a human, comprising administering to the human a therapeutically effective amount of the dimer of claim 37.

43. A method of treating painful diabetic neuropathy in a human, comprising administering to the human a therapeutically effective amount of the dimer of claim 37.

44. A composition comprising the dimer of claim 38 and a pharmaceutically acceptable carrier.

45. A method of treating neuropathic pain in a human, comprising administering to the human a therapeutically effective amount of the dimer of claim 38.

46. A method of activating the RET receptor in a human, comprising administering to the human an effective amount of the dimer of claim 38.

47. A method of treating a peripheral neuropathy in a human, comprising administering to the human a therapeutically effective amount of the dimer of claim 38.

48. A method of treating painful diabetic neuropathy in a human, comprising administering to the human a therapeutically effective amount of the dimer of claim 38.