Long lasting synthetic exendin-4 peptide conjugates and methods of use thereof

• US 20090075890A1
• Filed: 08/01/2008
• Published: 03/19/2009
• Est. Priority Date: 05/17/1999
• Status: Abandoned Application

• Current Assignee: ConjuChem LLC (Colabor Group, Inc.)
• Original Assignee: ConjuChem Biotechnologies, Inc. /Old/ (Colabor Group, Inc.)
• Inventors: Leblanc, Anouk, L'Archeveque, Benoit, Pierre, Serge St., Holmes, Darren L., Bridon, Dominique P.
• Application Number:
  US12/221,553
• Publication Number:
  US 20090075890A1
• Time in Patent Office: Days
• US Class Current: 514/1.100
• 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 47/643 Albumins, e.g. HSA, BSA, ovalbumin or a Keyhole Limpet Hemocyanin [KHL]
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 14/57563 Vasoactive intestinal peptide [VIP]; Related peptides
• CPC Class Codes: C07K 14/605 Glucagons
• CPC Class Codes: C12N 2740/16122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Abstract

Modified insulinotropic peptides are disclosed. The modified insulinotropic peptides are capable of forming a peptidase stabilized insulinotropic peptide. The modified insulinotropic peptides are capable of forming covalent bonds with one or more blood components to form a conjugate. The conjugates may be formed in vivo or ex vivo. The modified peptides are administered to treat humans with diabetes and other related diseases.

Claims

1-19. -19. (canceled)

20. A modified peptide comprising:

a peptide; and

a maleimide or a maleimido-containing reactive group coupled, optionally via a linker, to the peptide,

the modified peptide being capable of covalently binding in vivo or in vitro by reaction of the reactive group, with a functionality on a blood component;

wherein the peptide comprises exendin-4 or a C-terminal amide thereof.

21. The modified peptide of claim 20, wherein the amino acid sequence of the peptide is that of exendin-4(1-39) or a C-terminal amide thereof.

22. The modified peptide of claim 20, wherein the amino acid sequence of the peptide is that of exendin-4(1-39) Lys⁴⁰ or a C-terminal amide thereof.

23. The modified peptide of claim 22 which is according to the following formula:

24. The modified peptide of any one of claims 20-23, wherein the blood component is albumin.

25. The modified peptide of claim 20, wherein the functionality on the blood component is an amino group, an hydroxyl group, or a thiol group.

43. A pharmaceutical composition comprising the modified peptide of any of claims 20 to 25 and a physiologically acceptable medium.

49. A method of preparing a conjugate comprising:

contacting a modified peptide of claim 20 with a blood component under conditions suitable for formation of a conjugate between said modified peptide and said blood component.

50. The method of claim 49, wherein the amino acid sequence of the modified peptide is that of exendin-4(1-39) or a C-terminal amide thereof.

51. The method of claim 49, wherein the amino acid sequence of the modified peptide is that of exendin-4(1-39) Lys⁴⁰ or a C-terminal amide thereof.

52. The method of claim 49, wherein the reactive group of the modified peptide is a maleimido-containing group.

53. The method of claim 49, wherein the blood component is albumin.

54. The method of claim 53, wherein said modified peptide is conjugated to Cys-34 of human serum albumin.

55. The method of claim 49, wherein the contacting occurs in vitro.

56. The method of claim 49, wherein the modified peptide is according to the following formula:

57. A method for treating diabetes in a subject with diabetes, comprising administering to the subject a therapeutically effective amount of the modified peptide of claim 23.

58. The method of claim 57, wherein said modified compound is in association with a physiologically acceptable medium.

63. The method of any one of claims 57 to 62, wherein the diabetes is Type II diabetes.

64. The method of any one of claims 57 to 62, wherein the subject is a human.

65. A method for enhancing the expression of insulin in a subject, comprising administering to the subject a therapeutically effective amount of the modified peptide of claim 23.

66. The method of claim 65, wherein said modified peptide is in association with a physiologically acceptable medium.

71. The method of any one of claims 65 to 70, wherein the subject is a human.

72. A method for treating obesity in an obese subject, comprising administering to the subject a therapeutically effective amount of the modified peptide of claim 23.

73. The method of claim 72, wherein said modified peptide is in association with a physiologically acceptable medium.

78. The method of any one of claims 72 to 77, wherein the subject is a human.

79. A method for treating decreased insulin sensitivity in a subject with decreased insulin sensitivity, comprising administering to the subject a therapeutically effective amount of the modified peptide of claim 23.

80. The method of claim 79, wherein said modified peptide is in association with a physiologically acceptable medium.

85. The method of any one of claims 79 to 84, wherein the subject is a human.

26. A conjugate formed by conjugating

(a) a peptide coupled, optionally via a linker, to a maleimide or a maleimido-containing reactive group, with

(b) a blood component, by covalent linking of the reactive group with a functionality on the blood component,

wherein the peptide comprises exendin-4 or a C-terminal amide thereof.

27. The conjugate of claim 26, wherein the amino acid sequence of the peptide is that of exendin-4(1-39) or a C-terminal amide thereof.

28. The conjugate of claim 26, wherein the amino acid sequence of the peptide is that of exendin-4(1-39) Lys⁴⁰ or a C-terminal amide thereof.

29. The conjugate of claim 26 which is according to the formula:

44. A pharmaceutical composition comprising the conjugate of any of claims 26 to 42 and a physiologically acceptable medium.

30. A conjugate according to the formula:

31. The conjugate of claim 30, wherein the amino acid sequence of the peptide is that of exendin-4(1-39) or a C-terminal amide thereof.

32. The conjugate of claim 30, wherein the amino acid sequence of the peptide is that of exendin-4(1-39) Lys⁴⁰ or a C-terminal amide thereof.

33. The conjugate of claim 30, wherein L is present.

34. The conjugate of claim 33, wherein Z is covalently attached to L through the NH of an amino acid of Z.

35. The conjugate of claim 33, wherein L is a poly ethoxy amino acid.

36. The conjugate of claim 35, wherein the poly ethoxy amino acid is according to the formula:

—

[C(O)—

CH₂—

O—

(CH₂)₂—

NH]_m—

wherein m is 1, 2 or 3.

37. The conjugate of claim 35, wherein the poly ethoxy amino acid is according to the formula:

—

[C(O)—

CH₂—

O—

(CH₂)₂—

O—

(CH₂)₂—

NH]_m—

wherein m is 1, 2 or 3.

38. The conjugate of claim 30, wherein X is S.

39. The conjugate of claim 30, wherein the blood component is albumin.

40. The conjugate of claim 38, wherein the S is of Cys-34 of human serum albumin.

41. A conjugate according to the following formula:

42. The conjugate of claim 41, wherein said albumin is human serum albumin.

59. A method for treating diabetes in a subject with diabetes, comprising administering to the subject a therapeutically effective amount of the conjugate of claim 41.

60. The method of claim 59, wherein said albumin is human serum albumin.

61. The method of claim 59, wherein said conjugate is in association with a physiologically acceptable medium.

62. The method of claim 60, wherein said conjugate is in association with a physiologically acceptable medium.

67. A method for enhancing the expression of insulin in a subject, comprising administering to the subject a therapeutically effective amount of the conjugate of claim 41.

68. The method of claim 67, wherein said albumin is human serum albumin.

69. The method of claim 67, wherein said conjugate is in association with a physiologically acceptable medium.

70. The method of claim 68, wherein said conjugate is in association with a physiologically acceptable medium.

74. A method for treating obesity in an obese subject, comprising administering to the subject a therapeutically effective amount of the conjugate of claim 41.

75. The method of claim 74, wherein said albumin is human serum albumin.

76. The method of claim 74, wherein said conjugate is in association with a physiologically acceptable medium.

77. The method of claim 75, wherein said conjugate is in association with a physiologically acceptable medium.

81. A method for treating decreased insulin sensitivity in a subject with decreased insulin sensitivity, comprising administering to the subject a therapeutically effective amount of the conjugate of claim 41.

82. The method of claim 81, wherein said albumin is human serum albumin.

83. The method of claim 81, wherein said conjugate is in association with a physiologically acceptable medium.

84. The method of claim 82, wherein said conjugate is in association with a physiologically acceptable medium.

45. A method of synthesizing a modified exendin-4(1-39) peptide comprising:

(a) synthesizing a peptide from the carboxy terminal amino acid, wherein the amino acid sequence of the peptide is that of exendin-4(1-39); and

(b) adding a reactive group, optionally via a linking group, to the carboxy terminal amino acid.

47. The method of claim 45 or 46, wherein the reactive group is maleimide or a maleimido-containing group.

48. The method of claim 45 or 46, wherein the reactive group is succinimide or a succinimido-containing group.

46. A method of synthesizing a modified exendin-4(1-39) peptide comprising:

(a) synthesizing a peptide from the carboxy terminal amino acid, wherein the amino acid sequence of the peptide is that of exendin-4(1-39) Lys⁴⁰; and

(b) adding a reactive group, optionally via a linking group, to the carboxy terminal amino acid.