GIP AND GLP-1 RECEPTOR DUAL-AGONISTS FOR THE TREATMENT OF DIABETES

• US 20160185837A1
• Filed: 08/15/2014
• Published: 06/30/2016
• Est. Priority Date: 08/16/2013
• Status: Abandoned Application

• Current Assignee: Medimmmune Limited
• Original Assignee: MedImmune Ltd. (Astrazeneca PLC)
• Inventors: Collinson, Andie, Hornigold, David, O'Brien, Siobhan, Papworth, Monika, Ravn, Peter, Sermadiras, Isabelle, Bednarek, Maria
• Application Number:
  US14/911,910
• Publication Number:
  US 20160185837A1
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/54 characterised by the route of administration
• CPC Class Codes: A61K 38/2235 Secretins
• CPC Class Codes: A61K 38/26 Glucagons
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: C07K 14/57563 Vasoactive intestinal peptide [VIP]; Related peptides
• CPC Class Codes: C07K 14/605 Glucagons
• CPC Class Codes: C07K 14/645 Secretins
• CPC Class Codes: C07K 16/26 against hormones ; against hormone releasing or inhibiting factors
• CPC Class Codes: C07K 2317/524 CH2 domain
• CPC Class Codes: C07K 2317/526 CH3 domain
• CPC Class Codes: C07K 2317/71 Decreased effector function due to an Fc-modification
• CPC Class Codes: C07K 2317/75 Agonist effect on antigen
• CPC Class Codes: C07K 2317/94 Stability, e.g. half-life, pH, temperature or enzyme-resistance
• 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

Abstract

This disclosure provides GIP/GLP-1 dual agonist polypeptides for the treatment of hypoglycemic conditions, e.g., type-2 diabetes.

Claims

1. An isolated polypeptide comprising the amino acid sequence:

2. The polypeptide of claim 1, wherein X₁₃ is L.

3. The polypeptide of claim 2, wherein X₃ is E, X₁₀ is Y, X₁₂ is K, X₁₄ is L, X₁₅ is E, X₁₆ is E, X₁₇ is E, X₁₈ is A, X₁₉ is V, X₂₀ is R, X₂₁ is L, X₂₃ is I;

X₂₄ is E; and

X₂₉ is G.

4. The polypeptide of any one of claims 1 to 3, wherein X₂₇ is L and X₂₈ is A;

X₂₇ is K and X₂₈ is N;

or X₂₇ is V and X₂₈ is K.

5. The polypeptide of claim 1, comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

2-32.

26. The polypeptide of any one of claims 1 to 25, further comprising a heterologous polypeptide fused thereto.

27. The polypeptide of claim 26, wherein the heterologous polypeptide comprises a linker, a hinge, an Fc domain, or a combination thereof.

28. The polypeptide of claim 27, wherein the linker comprises (GGGGS)n, wherein n 1, 2, 3, or 4.

29. The polypeptide of claim 27 or claim 28, wherein the linker comprises the amino acid sequence:

G GGGGS GGGGS GGGGS GGGGS A (SEQ ID NO;

66), A PPGGS GGGGS GGGGS GGGGS A (SEQ ID NO;

67), GT GGGGS GGGGS GGGGS GGGGS A (SEQ ID NO;

68), G GGGGS GGGGS GGGGS GGGGS A (SEQ ID NO;

69), G GGGGS A (SEQ ID NO;

70), G GGGGS GGGGS A (SEQ ID NO;

71), G GGGGS GGGGS GGGGS A (SEQ ID NO;

72), G KGGGS GGGGS GGGGS GGGGS A (SEQ ID NO;

73), G GGGGS GGGGS GGGGS GGGGSA (SEQ ID NO;

179), G GGGG GGGG GGGG GGGG A (SEQ ID NO;

180) or any combination thereof.

30. The polypeptide of claim 27, wherein the linker comprises GG SGSTA SSGSG SATGG GGAA (SEQ ID NO:

74), any variant thereof, or any fragment thereof.

31. The polypeptide of claim 30, wherein the linker comprises AAAGG SGSTA SSGSG SATGG GGAA (SEQ ID NO:

75), APPGG SGSTA SSGSG SATGG GGAA (SEQ ID NO;

76), or any combination thereof.

32. The polypeptide of claim 27, wherein the hinge comprises an amino acid sequence of an IgG1 hinge, an IgG4 hinge, a fragment thereof, a variant thereof, or any combination thereof.

33. The polypeptide of claim 32, wherein the hinge comprises ESKYGPPCPPCPAPEAA (SEQ ID NO:

77), THTCPPCPAPEF (SEQ ID NO;

78), THTCPPC (SEQ ID NO;

79), CPPCPAPEF (SEQ ID NO;

80), TYTCPPCPAPEF (SEQ ID NO;

81), TSTCPPCPAPEF (SEQ ID NO;

82), PPCPPCPAPEF (SEQ ID NO;

83), ESKYGPPCPPCPAPEF (SEQ ID NO;

84), APEF (SEQ ID NO;

85), ESKYGPPCPPC (SEQ ID NO;

86), THTCPPCPAPELL (SEQ ID NO;

87), any variant thereof, any fragment thereof, or any combination thereof.

34. The polypeptide of claim 27, wherein the Fc region comprises an IgG1 Fc region, an IgG1-TM Fc region, an IgG1-FQQ Fc region, an IgG4 Fc region, an IgG1-YTE Fc region, any fragment thereof, any variant thereof, or any combination thereof.

35. The polypeptide of claim any one of claims 26 to 34, comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

88-175 and 326-460.

36. The polypeptide of any one of claims 1 to 35, further comprising a heterologous moiety attached thereto.

37. The polypeptide of claim 36, wherein the heterologous moiety is a polypeptide, an organic polymer, an inorganic polymer, a polyethylene glycol (PEG), biotin, an albumin, a human serum albumin (HSA), a HSA FcRn binding portion, an antibody, a domain of an antibody, an antibody fragment, a single chain antibody, a domain antibody, an albumin binding domain, an enzyme, a ligand, a receptor, a binding peptide, a non-FnIII scaffold, an epitope tag, a recombinant polypeptide polymer, a cytokine, or a combination of two or more of the recited moieties.

38. The polypeptide of any one of claims 1 to 37, wherein the peptide binds to a GIP receptor, binds to a GLP-1 receptor, or binds to both a GIP and a GLP-1 receptor.

39. The polypeptide of claim 38, which binds to a GIP receptor.

40. The polypeptide of claim 39, wherein the GIP receptor is a mouse GIP receptor, a rat GIP receptor, or a human GIP receptor.

41. The polypeptide any one of claims 38 to 40, which binds to a human GIP receptor with an EC50 in the cAMP assay 1 of less than 10,000 pM, less than 5000 pM, less than 2500 pM, less than 1000 pM, less than 900 pM, less than 800 pM, less than 700 pM, less than 600 pM, less than 500 pM, less than 400 pM, less than 300 pM, less than 200 pM, less than 100 pM, less than 50 pM, less than 25 pM, less than 20 pM, less than 15 pM, less than 10 pM, less than 5 pM, less than 4 pM, less than 3 pM, or less than 2 pM.

42. The polypeptide of any one of claims 38 to 41, which binds to a human glucagon receptor with at least 1000-fold lower affinity than its binding affinity for a human GIP receptor, as measured in the cAMP assay.

43. The polypeptide of any one of claims 38 to 42, which binds to a GLP-1 receptor.

44. The polypeptide of claim 43, wherein the GLP-1 receptor is a mouse GLP-1 receptor, a rat GLP-1 receptor, or a human GLP-1 receptor.

45. The polypeptide of claim 43 or claim 44, which binds to a human GLP-1 receptor with an EC50 in the cAMP assay 1 of less than 10,000 pM, less than 5000 pM, less than 2500 pM, less than 1000 pM, less than 900 pM, less than 800 pM, less than 700 pM, less than 600 pM, less than 500 pM, less than 400 pM, less than 300 pM, less than 200 pM, less than 100 pM, less than 50 pM, less than 25 pM, less than 20 pM, less than 15 pM, less than 10 pM, less than 5 pM, less than 4 pM, less than 3 pM, or less than 2 pM.

46. The polypeptide of any one of claims 43 to 45, which binds to a human glucagon receptor with at least 1000-fold lower affinity than its binding affinity for a human GLP-1 receptor, as measured in the cAMP assay.

47. The polypeptide of any one of claims 38 to 46, which is an agonist of GLP-1 activity, an agonist of GIP activity, or an agonist of both GLP-1 and GIP activity.

48. An isolated polynucleotide encoding the polypeptide of any one of claims 1 to 47.

49. A vector comprising the polynucleotide of claim 48.

50. A host cell comprising the polynucleotide of claim 48 or the vector of claim 49.

51. A method of making the polypeptide of any one of claims 1 to 47, comprising culturing the host cell of claim 50 under conditions allowing expression of the polypeptide, and recovering the polypeptide.

52. A pharmaceutical composition comprising the polypeptide of any one of claims 1 to 47, and a carrier.

53. A kit comprising the composition of claim 52.

54. A method of treating or preventing a disease or condition caused or characterized by hypoglycemia or impaired insulin release, comprising administering to a subject in need of treatment an effective amount of the polypeptide of any one of claims 1 to 47 or the composition of claim 52.

55. The method of claim 54, wherein the disease or condition is diabetes.

56. The method of claim 55, wherein the disease or condition is type-2 diabetes.

57. The method of any one of claims 54 to 56, wherein the administration further improves glycemic control, provides body weight control, improves β

-cell function and mass, reduces the rate of gastric acid secretion and gastric emptying, or any combination thereof.

58. The method of any one of claims 54 to 57, wherein the peptide is administered orally or by injection.

59. The method of claim 58, wherein the injection is administered subcutaneously or intravenously.

60. The method of claim 58 or claim 59, wherein the polypeptide is administered once per day.

61. The method of any one of claims 54 to 60, further comprising administering one or more additional therapies.

62. The method of claim 61, wherein the additional therapy comprises blood sugar monitoring, diet modifications, exercise, insulin, a thiazolidinedione, a sulfonylurea, an incretin, metformin, a glyburide, a dipeptidyl peptidase 4 inhibitor, a bile acid sequestrant, or any combination thereof.

63. The method of any one of claims 54 to 62, wherein the subject is human.

6. An isolated polypeptide comprising the amino acid sequence:

7. The polypeptide of claim 6, wherein X₁ is Y and X₂ is S.

8. The polypeptide of any one of claim 6 or claim 7, wherein X₁₀ is I, X₁₁ is A, and X₁₂ is W.

9. The polypeptide of any one of claims 6 to 8, wherein X₁₄ is G and X₁₅ is no amino acid or G.

10. The polypeptide of any one of claims 6 to 9, wherein X₇ is E.

11. The polypeptide of any one of claims 6 to 10, wherein X₈ is R and X₉ is Q.

12. The polypeptide of any one of claims 6 to 11, wherein X₁₃ is K.

13. The polypeptide of any one of claims 6 to 12, wherein X₃ is V.

14. The polypeptide of any one of claims 6 to 14, wherein X₄ is S, X₅ is Y, and X₆ is M.

15. The polypeptide of claim 6, comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

33-43.

16. An isolated polypeptide comprising the amino acid sequence of IP171, IP176, IP206, IP214, IP215, IP175. IP208, IP204, IP199, IP173, IP150, IP174, IP205, IP127, or IP200.

17. An isolated polypeptide comprising the amino acid sequence:

18. The polypeptide of claim 17, comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

2-32, and 191-256.

19. An isolated polypeptide comprising the amino acid sequence:

20. The polypeptide of claim 19, comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

33-43 and 257-294.

21. An isolated polypeptide comprising the amino acid sequence:

22. The polypeptide of claim 21, wherein X₄ is L.

23. The polypeptide of claim 22, wherein X₁ is E;

X₂ is Y;

X₃ is K;

X₅ is L;

X₆ is E;

X₇ is E;

X₈ is E;

X₉ is A;

X₁₀ is V;

X₁₁ is R, X₁₂ is L;

X₁₃ is I;

X₁₄ is E; and

X₁₇ is G.

24. The polypeptide of any one of claims 21 to 23, wherein X₁₅ is L and X₁₆ is A;

X₁₅ is K and X₁₆ is N;

or X₁₅ is V and X₁₆ is K.

25. The polypeptide of claim 21, comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

2-32.