Polymer conjugates of mutated neublastin

• US 8,119,114 B2
• Filed: 06/27/2008
• Issued: 02/21/2012
• Est. Priority Date: 02/01/2001
• Status: Expired due to Fees

• Current Assignee: Gloriana Therapeutics, Inc.
• Original Assignee: Biogen Idec MA Inc. (Biogen, Inc.)
• Inventors: Sah, Dinah Wen-Yee, Pepinsky, R. Blake, Boriack-Sjodin, Paula Ann, Miller, Stephan S., Rossomando, Anthony, Silvian, Laura
• Primary Examiner(s): Kolker, Daniel E
• Assistant Examiner(s): Emch, Gregory S
• Application Number:
  US12/163,425
• Publication Number:
  US 20080306212A1
• Time in Patent Office: 1,334 Days
• Field of Search: None
• US Class Current: 424/78.18
• 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: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 25/04 Centrally acting analgesics, e.g. opioids
• CPC Class Codes: C07K 14/475 Growth factors; Growth regulators
• CPC Class Codes: C07K 14/4756 Neuregulins, i.e. p185erbB2 ligands, glial growth factor, heregulin, ARIA, neu differentiation factor
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

A dimer comprising a mutated neublastin polypeptide coupled to a polymer is disclosed. Such dimers exhibit prolonged bioavailability and, in preferred embodiments, prolonged biological activity relative to wild-type forms of neublastin.

Claims

1. A polymer-conjugated homodimer comprising two polypeptides, wherein each polypeptide comprises an amino acid sequence at least 90% identical to amino acids 8-113 of SEQ ID NO:

1, wherein the polypeptide comprises an amino acid other than arginine substituted at the position corresponding to position 14 in SEQ ID NO;

1, wherein the polymer-conjugated homodimer binds to GFRalpha3 and stimulates tyrosine phosphorylation of RET receptor tyrosine kinase, and wherein a non-naturally occurring polymer is conjugated to the amino acid substituted at the position corresponding to position 14 in SEQ ID NO;

1.

2. The polymer-conjugated homodimer of claim 1, wherein the amino acid sequence is at least 95% identical to amino acids 8-113 of SEQ ID NO:

1.

3. The polymer-conjugated homodimer of claim 1, wherein the amino acid sequence is at least 95% identical to amino acids 8-113 of SEQ ID NO:

2.

4. The polymer-conjugated homodimer of claim 1, wherein the polymer is a polyalkylene glycol.

5. The polymer-conjugated homodimer of claim 2, wherein the polymer is a polyalkylene glycol.

6. The polymer-conjugated homodimer of claim 3, wherein the polymer is a polyalkylene glycol.

7. The polymer-conjugated homodimer of claim 4, wherein the polyalkylene glycol is conjugated to a lysine residue substituted at the position corresponding to position 14 in SEQ ID NO:

1.

8. The polymer-conjugated homodimer of claim 5, wherein the polyalkylene glycol is conjugated to a lysine residue substituted at the position corresponding to position 14 in SEQ ID NO:

1.

9. The polymer-conjugated homodimer of claim 6, wherein the polyalkylene glycol is conjugated to a lysine residue substituted at the position corresponding to position 14 in SEQ ID NO:

1.

10. The polymer-conjugated homodimer of claim 4, wherein the polyalkylene glycol is polyethylene glycol.

11. The polymer-conjugated homodimer of claim 5, wherein the polyalkylene glycol is polyethylene glycol.

12. The polymer-conjugated homodimer of claim 6, wherein the polyalkylene glycol is polyethylene glycol.

13. The polymer-conjugated homodimer of claim 7, wherein the polyalkylene glycol is polyethylene glycol.

14. The polymer-conjugated homodimer of claim 8, wherein the polyalkylene glycol is polyethylene glycol.

15. The polymer-conjugated homodimer of claim 9, wherein the polyalkylene glycol is polyethylene glycol.

16. A polymer-conjugated homodimer comprising two polypeptides, wherein each polypeptide comprises an amino acid sequence at least 90% identical to amino acids 8-113 of SEQ ID NO:

1, wherein the polypeptide comprises an amino acid other than arginine substituted at the position corresponding to position 39 in SEQ ID NO;

1, wherein the polymer-conjugated homodimer binds to GFRalpha3 and stimulates tyrosine phosphorylation of RET receptor tyrosine kinase, and wherein a non-naturally occurring polymer is conjugated to the amino acid substituted at the position corresponding to position 39 in SEQ ID NO;

1.

17. The polymer-conjugated homodimer of claim 16, wherein the amino acid sequence is at least 95% identical to amino acids 8-113 of SEQ ID NO:

1.

18. The polymer-conjugated homodimer of claim 16, wherein the amino acid sequence is at least 95% identical to amino acids 8-113 of SEQ ID NO:

2.

19. The polymer-conjugated homodimer of claim 16, wherein the polymer is a polyalkylene glycol.

20. The polymer-conjugated homodimer of claim 17, wherein the polymer is a polyalkylene glycol.

21. The polymer-conjugated homodimer of claim 18, wherein the polymer is a polyalkylene glycol.

22. The polymer-conjugated homodimer of claim 19, wherein the polyalkylene glycol is conjugated to a lysine residue substituted at the position corresponding to position 39 in SEQ ID NO:

1.

23. The polymer-conjugated homodimer of claim 20, wherein the polyalkylene glycol is conjugated to a lysine residue substituted at the position corresponding to position 39 in SEQ ID NO:

1.

24. The polymer-conjugated homodimer of claim 21, wherein the polyalkylene glycol is conjugated to a lysine residue substituted at the position corresponding to position 39 in SEQ ID NO:

1.

25. The polymer-conjugated homodimer of claim 19, wherein the polyalkylene glycol is polyethylene glycol.

26. The polymer-conjugated homodimer of claim 20, wherein the polyalkylene glycol is polyethylene glycol.

27. The polymer-conjugated homodimer of claim 21, wherein the polyalkylene glycol is polyethylene glycol.

28. The polymer-conjugated homodimer of claim 22, wherein the polyalkylene glycol is polyethylene glycol.

29. The polymer-conjugated homodimer of claim 23, wherein the polyalkylene glycol is polyethylene glycol.

30. The polymer-conjugated homodimer of claim 24, wherein the polyalkylene glycol is polyethylene glycol.

31. A polymer-conjugated homodimer comprising two polypeptides, wherein each polypeptide comprises an amino acid sequence at least 90% identical to amino acids 8-113 of SEQ ID NO:

1, wherein the polypeptide comprises an amino acid other than arginine substituted at the position corresponding to position 68 in SEQ ID NO;

1, wherein the polymer-conjugated homodimer binds to GFRalpha3 and stimulates tyrosine phosphorylation of RET receptor tyrosine kinase, and wherein a non-naturally occurring polymer is conjugated to the amino acid substituted at the position corresponding to position 68 in SEQ ID NO;

1.

32. The polymer-conjugated homodimer of claim 31, wherein the amino acid sequence is at least 95% identical to amino acids 8-113 of SEQ ID NO:

1.

33. The polymer-conjugated homodimer of claim 31, wherein the amino acid sequence is at least 95% identical to amino acids 8-113 of SEQ ID NO:

2.

34. The polymer-conjugated homodimer of claim 31, wherein the polymer is a polyalkylene glycol.

35. The polymer-conjugated homodimer of claim 32, wherein the polymer is a polyalkylene glycol.

36. The polymer-conjugated homodimer of claim 33, wherein the polymer is a polyalkylene glycol.

37. The polymer-conjugated homodimer of claim 34, wherein the polyalkylene glycol is conjugated to a lysine residue substituted at the position corresponding to position 68 in SEQ ID NO:

1.

38. The polymer-conjugated homodimer of claim 35, wherein the polyalkylene glycol is conjugated to a lysine residue substituted at the position corresponding to position 68 in SEQ ID NO:

1.

39. The polymer-conjugated homodimer of claim 36, wherein the polyalkylene glycol is conjugated to a lysine residue substituted at the position corresponding to position 68 in SEQ ID NO:

1.

40. The polymer-conjugated homodimer of claim 34, wherein the polyalkylene glycol is polyethylene glycol.

41. The polymer-conjugated homodimer of claim 35, wherein the polyalkylene glycol is polyethylene glycol.

42. The polymer-conjugated homodimer of claim 36, wherein the polyalkylene glycol is polyethylene glycol.

43. The polymer-conjugated homodimer of claim 37, wherein the polyalkylene glycol is polyethylene glycol.

44. The polymer-conjugated homodimer of claim 38, wherein the polyalkylene glycol is polyethylene glycol.

45. The polymer-conjugated homodimer of claim 39, wherein the polyalkylene glycol is polyethylene glycol.

46. A polymer-conjugated homodimer comprising two polypeptides, wherein each polypeptide comprises an amino acid sequence at least 90% identical to amino acids 8-113 of SEQ ID NO:

1, wherein the polypeptide comprises an amino acid other than asparagine substituted at the position corresponding to position 95 in SEQ ID NO;

1, wherein the polymer-conjugated homodimer binds to GFRalpha3 and stimulates tyrosine phosphorylation of RET receptor tyrosine kinase, and wherein a non-naturally occurring polymer is conjugated to the amino acid substituted at the position corresponding to position 95 in SEQ ID NO;

1.

47. The polymer-conjugated homodimer of claim 46, wherein the amino acid sequence is at least 95% identical to amino acids 8-113 of SEQ ID NO:

1.

48. The polymer-conjugated homodimer of claim 46, wherein the amino acid sequence is at least 95% identical to amino acids 8-113 of SEQ ID NO:

2.

49. The polymer-conjugated homodimer of claim 46, wherein the polymer is a polyalkylene glycol.

50. The polymer-conjugated homodimer of claim 47, wherein the polymer is a polyalkylene glycol.

51. The polymer-conjugated homodimer of claim 48, wherein the polymer is a polyalkylene glycol.

52. The polymer-conjugated homodimer of claim 49, wherein the polyalkylene glycol is conjugated to a lysine residue substituted at the position corresponding to position 95 in SEQ ID NO:

1.

53. The polymer-conjugated homodimer of claim 50, wherein the polyalkylene glycol is conjugated to a lysine residue substituted at the position corresponding to position 95 in SEQ ID NO:

1.

54. The polymer-conjugated homodimer of claim 51, wherein the polyalkylene glycol is conjugated to a lysine residue substituted at the position corresponding to position 95 in SEQ ID NO:

1.

55. The polymer-conjugated homodimer of claim 49, wherein the polyalkylene glycol is polyethylene glycol.

56. The polymer-conjugated homodimer of claim 50, wherein the polyalkylene glycol is polyethylene glycol.

57. The polymer-conjugated homodimer of claim 51, wherein the polyalkylene glycol is polyethylene glycol.

58. The polymer-conjugated homodimer of claim 52, wherein the polyalkylene glycol is polyethylene glycol.

59. The polymer-conjugated homodimer of claim 53, wherein the polyalkylene glycol is polyethylene glycol.

60. The polymer-conjugated homodimer of claim 54, wherein the polyalkylene glycol is polyethylene glycol.