PEGYLATED OXM VARIANTS

• US 20150148289A1
• Filed: 06/04/2013
• Published: 05/28/2015
• Est. Priority Date: 06/04/2012
• Status: Active Grant

• Current Assignee: Opko Biologics Ltd. (Opko Health, Inc.)
• Original Assignee: Opko Biologics Ltd. (Opko Health, Inc.)
• Inventors: Fima, Udi Eyal, Hershkovitz, Oren
• Granted Patent: US 10,537,646 B2
• Time in Patent Office: Days
• US Class Current: 514/5.3
• CPC Class Codes: A61K 38/22 Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin A61K38/33, e.g. corticotropin A61K38/35
• CPC Class Codes: A61K 38/26 Glucagons
• 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 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C07K 1/00 General methods for the preparation of peptides , i.e. processes for the organic chemical preparation of peptides or proteins of any length
• CPC Class Codes: C07K 14/605 Glucagons

Abstract

A composition which includes oxyntomodulin and polyethylene glycol polymer (PEG polymer) linked via a reversible linker such as 9-fluorenylmethoxycarbonyl (Fmoc) or 2-sulfo-9-fluorenylmethoxycarbonyl (FMS) is disclosed. Pharmaceutical compositions comprising the reverse pegylated oxyntomodulin and methods of using same are also disclosed.

Claims

1-43. -43. (canceled)

44. A composition consisting of an oxyntomodulin, a polyethylene glycol polymer (PEG polymer) and 9-fluorenylmethoxycarbonyl (Fmoc) or 2-sulfo-9-fluorenylmethoxycarbonyl (FMS), wherein said PEG polymer is attached to the amino terminus of said oxyntomodulin or to a lysine residue on position number twelve (Lys₁₂) or to a lysine residue on position number thirty (Lys₃₀) of said oxyntomodulin, via Fmoc or FMS.

45. The composition of claim 44, wherein said oxyntomodulin consists of the amino acid sequence set forth in SEQ ID NO:

1.

46. The composition of claim 44, wherein said PEG polymer is a PEG polymer with a sulfhydryl moiety.

47. The composition of claim 44, wherein said PEG polymer is PEG₃₀.

48. A pharmaceutical composition comprising the composition of claim 44 and a pharmaceutical acceptable carrier.

69. A method of inducing glucose tolerance in a subject in need thereof, or increasing insulin sensitivity in a subject in need thereof, or inducing glycemic control in a subject in need thereof, or reducing insulin resistance in a subject in need thereof, the method comprising administering the pharmaceutical composition of claim 48.

70. A method of reducing food intake in a subject in need thereof, or reducing body weight in a subject in need thereof, or for increasing energy expenditure in a subject in need thereof, or improving the cholesterol levels in a subject in need thereof, the method comprising administering the pharmaceutical composition of claim 48.

49. A method of improving the area under the curve (AUC) of oxyntomodulin, consisting of the step of conjugating a polyethylene glycol polymer (PEG polymer) to the Lysine residue on position number 12 or to the Lysine residue on position number 30 or to the amino terminus of said oxyntomodulin amino acid sequence via 9-fluorenylmethoxycarbonyl (Fmoc) or 2-sulfo-9-fluorenylmethoxycarbonyl (FMS).

50. The method of claim 49, wherein said oxyntomodulin consists of an amino acid sequence of SEQ ID NO:

1.

51. The method of claim 49, wherein said PEG polymer is conjugated to the amino terminus or lysine residue of said oxyntomodulin via Fmoc or FMS.

52. The method of claim 49, wherein said PEG polymer is a PEG polymer with a sulfhydryl moiety.

53. The method of claim 49, wherein said PEG polymer is PEG₃₀.

54. A method of reducing the dosing frequency of oxyntomodulin, consisting of the step of conjugating a polyethylene glycol polymer (PEG polymer) to the Lysine residue on position number 12 or to the Lysine residue on position number 30 or to the amino terminus of said oxyntomodulin amino acid sequence via 9-fluorenylmethoxycarbonyl (Fmoc) or 2-sulfo-9-fluorenylmethoxycarbonyl (FMS).

55. The method of claim 54, wherein said oxyntomodulin consists of an amino acid sequence of SEQ ID NO:

1.

56. The method of claim 54, wherein said PEG polymer is a PEG polymer with a sulfhydryl moiety.

57. The method of claim 54, wherein said PEG polymer is PEG₃₀.

58. A method for extending the biological half-life of oxyntomodulin, consisting of the step of conjugating oxyntomodulin, a polyethylene glycol polymer (PEG polymer) and 9-fluorenylmethoxycarbonyl (Fmoc) or 2-sulfo-9-fluorenylmethoxycarbonyl (FMS), wherein said PEG polymer is conjugated to a Lysine residue on position number 12 or to a Lysine residue on position number 30 or to the amino terminus of said oxyntomodulin amino acid sequence via 9-fluorenylmethoxycarbonyl (Fmoc) or 2-sulfo-9-fluorenylmethoxycarbonyl (FMS).

59. The method of claim 58, wherein said oxyntomodulin consists of an amino acid sequence of SEQ ID NO:

1.

60. The method of claim 58, wherein said PEG polymer is conjugated to the amino terminus or lysine residue of said oxyntomodulin via Fmoc or FMS.

61. The method of claim 58, wherein said PEG polymer is a PEG polymer with a sulfhydryl moiety.

62. The method of claim 58, wherein said PEG polymer is PEG₃₀.

63. The method of claim 58, wherein said oxyntomodulin is released into a biological fluid by chemically hydrolyzing said FMS or said Fmoc linker from said composition.

64. The method of claim 63, wherein the released oxyntomodulin is intact and regains GLP-1 and glucagon receptor binding activity.

65. The method of claim 63, wherein said biological fluid is blood, sera, or cerebrospinal fluid.

66. The method of claim 63, wherein said biological fluid is in a human subject.

67. The method of claim 63, wherein chemically hydrolyzing said FMS or said Fmoc extends the circulating time of said oxyntomodulin peptide in said biological fluid.

68. The method of claim 67, wherein extending the circulating time of said oxyntomodulin (OXM) allows said OXM to cross the blood brain barrier and target the CNS.