Conjugate comprising oxyntomodulin and an immunoglobulin fragment, and use thereof

• US 9,731,031 B2
• Filed: 06/15/2012
• Issued: 08/15/2017
• Est. Priority Date: 06/17/2011
• Status: Active Grant

• Current Assignee: Hanmi Science Co., Ltd
• Original Assignee: Hanmi Science Co., Ltd
• Inventors: Park, Sung Hee, Woo, Young Eun, Choi, In Young, Jung, Sung Youb, Kim, Dae Jin, Kwon, Se Chang
• Primary Examiner(s): Ha, Julie
• Assistant Examiner(s): KOMATSU, LI N
• Application Number:
  US14/126,914
• Publication Number:
  US 20140212440A1
• Time in Patent Office: 1,887 Days
• 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 38/26 Glucagons
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• 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: A61K 47/68 the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
• CPC Class Codes: A61K 47/6801 Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
• CPC Class Codes: A61K 47/6811 the drug being a protein or peptide, e.g. transferrin or bleomycin
• CPC Class Codes: A61K 47/6889 Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: C07K 14/575 Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695
• CPC Class Codes: C07K 14/605 Glucagons
• 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

The present invention relates to a conjugate comprising oxyntomodulin, an immunoglobulin Fc region, and non-peptidyl polymer wherein the conjugate being obtainable by covalently linking oxyntomodulin to immunoglobulin Fc region via non-peptidyl polymer, and a pharmaceutical composition for the prevention or treatment of obesity comprising the conjugates. The conjugate comprising oxyntomodulin and the immunoglobulin Fc of the present invention reduces food intake, suppresses gastric emptying, and facilitates lipolysis without side-effects, unlike native oxyntomodulin, and also shows excellent receptor-activating effects and long-term sustainability, compared to native oxyntomodulin. Thus, it can be widely used in the treatment of obesity with safety and efficacy.

Claims

1. A conjugate comprising:

an oxyntomodulin derivative comprising the amino acid sequence of SEQ ID NO;

24;

an immunoglobulin Fc region; and

a non-peptidyl polymer, wherein the non-peptidyl polymer links the oxyntomodulin derivative and the immunoglobulin Fc region via covalent bonds.

2. The conjugate according to claim 1, wherein the oxyntomodulin derivative activates glucagon-like peptide-1 receptor and glucagon receptor.

3. The conjugate according to claim 1, wherein the conjugate has anti-obesity effects.

4. The conjugate according to claim 1, wherein the non-peptidyl polymer is polyethylene glycol, polypropylene glycol, copolymers of ethylene glycol and propylene glycol, polyoxyethylated polyols, polyvinyl alcohol, polysaccharides, polyvinyl ethyl ether, polylactic acid, polylacticglycolic acid, lipid polymers, hyaluronic acid, or combinations thereof.

5. The conjugate according to claim 4, wherein the non-peptidyl polymer is polyethylene glycol.

6. The conjugate according to claim 4, wherein the polysaccharide is dextran, chitin, or a combination thereof.

7. The conjugate according to claim 1, wherein one functional group selected from the group consisting of an amine group and a thiol group of the immunoglobulin Fc region is attached to one end of the non-peptidyl polymer, and one functional group selected from the group consisting of an amine group and a thiol group of the oxyntomodulin derivative is attached to the other end of the non-peptidyl polymer.

8. The conjugate according to claim 1, wherein the conjugate is prepared by covalently linking the oxyntomodulin derivative to the immunoglobulin Fc region via the non-peptidyl polymer, the non-peptidyl polymer having, prior to forming the conjugate, reactive functional groups at both ends.

9. The conjugate according to claim 8, wherein the reactive functional groups are an aldehyde group, a maleimide group, a succinimide derivative, or combinations thereof.

10. The conjugate according to claim 1, wherein the immunoglobulin Fc region is a non-glycosylated Fc region.

11. The conjugate according to claim 1, wherein the immunoglobulin Fc region is selected from the group consisting of a heavy-chain constant region 1 (CH1) domain, a heavy-chain constant region 2 (CH2) domain, a heavy-chain constant region 3 (CH3) domain and a heavy-chain constant region 4 (CH4) domain;

a CH1 domain and a CH2 domain;

a CH1 domain and a CH3 domain;

a CH2 domain and a CH3 domain;

a combination of one or more domains and an immunoglobulin hinge region or a portion of the hinge region; and

a dimer of each domain of the heavy-chain constant regions and the light-chain constant region.

12. The conjugate according to claim 1, wherein the immunoglobulin Fc region comprises a modification of a native Fc region selected from the group consisting of:

(i) an amino acid modification to prevent formation of a disulfide bond;

(ii) elimination of one or more amino acid residues from the N-terminal end of the native Fc region;

(iii) addition of a methionine residue to the N-terminal end of the Fc region;

(iv) deletion of a complement-binding site; and

(v) deletion of an antibody dependent cell mediated cytotoxicity (ADCC) site.

13. The conjugate according to claim 1, wherein the immunoglobulin Fc region is an Fc region from an immunoglobulin selected from the group consisting of immunoglobulin G, immunoglobulin A, immunoglobulin D, immunoglobulin E, and immunoglobulin M.

14. The conjugate according to claim 13, wherein the immunoglobulin Fc region is an IgG4 Fc region.

15. The conjugate according to claim 1, wherein the immunoglobulin Fc region is a human IgG4-derived non-glycosylated Fc region.

16. The conjugate of claim 1, wherein the non-peptidyl polymer is linked via a covalent bond to the oxyntomodulin derivative at a cysteine residue.

17. A pharmaceutical composition comprising the conjugate of claim 1 and a pharmaceutically acceptable carrier.

18. The pharmaceutical composition according to claim 17, further comprising a glucagon-like peptide-1 receptor agonist, a leptin receptor agonist, a dipeptidyl peptidase-IV inhibitor, a Y5 receptor antagonist, a melanin-concentrating hormone receptor antagonist, a Y2/3 receptor agonist, a MC3/4 receptor agonist, a gastric/pancreatic lipase inhibitor, a 5HT2c agonist, a β

3A receptor agonist, an amylin receptor agonist, a ghrelin antagonist, or a ghrelin receptor antagonist.

19. A method for treating obesity, comprising administering the conjugate of claim 1 to a subject in need thereof.

20. The method of claim 19, which further comprises administering an additional drug selected from the group consisting of a glucagon-like peptide-1 receptor agonist, a leptin receptor agonist, a dipeptidyl peptidase-IV inhibitor, a Y5 receptor antagonist, a melanin-concentrating hormone receptor antagonist, a Y2/3 receptor agonist, a MC3/4 receptor agonist, a gastric/pancreatic lipase inhibitor, a 5HT2c agonist, a β

3A receptor agonist, an amylin receptor agonist, a ghrelin antagonist, and a ghrelin receptor antagonist.

21. A conjugate comprising:

an oxyntomodulin derivative comprising the amino acid sequence of SEQ ID NO;

25;

an immunoglobulin Fc region; and

a non-peptidyl polymer, wherein the non-peptidyl polymer links the oxyntomodulin derivative and the immunoglobulin Fc region via covalent bonds.

22. The conjugate according to claim 21, wherein the oxyntomodulin derivative activates glucagon-like peptide-1 receptor and glucagon receptor.

23. The conjugate according to claim 21, wherein the conjugate has anti-obesity effects.

24. The conjugate according to claim 21, wherein the non-peptidyl polymer is polyethylene glycol, polypropylene glycol, copolymers of ethylene glycol and propylene glycol, polyoxyethylated polyols, polyvinyl alcohol, polysaccharides, polyvinyl ethyl ether, polylactic acid, polylacticglycolic acid, lipid polymers, hyaluronic acid, or combinations thereof.

25. The conjugate according to claim 24, wherein the non-peptidyl polymer is polyethylene glycol.

26. The conjugate according to claim 24, wherein the polysaccharide is dextran, chitin, or a combination thereof.

27. The conjugate according to claim 21, wherein one functional group selected from the group consisting of an amine group and a thiol group of the immunoglobulin Fc region is attached to one end of the non-peptidyl polymer, and one functional group selected from the group consisting of an amine group and a thiol group of the oxyntomodulin derivative is attached to the other end of the non-peptidyl polymer.

28. The conjugate according to claim 21, wherein the conjugate is prepared by covalently linking the an oxyntomodulin derivative to the an immunoglobulin Fc region via the non-peptidyl polymer, the non-peptidyl polymer having, prior to forming the conjugate, reactive functional groups at both ends.

29. The conjugate according to claim 28, wherein the reactive functional groups are an aldehyde group, a maleimide group, a succinimide derivative, or combinations thereof.

30. The conjugate according to claim 21, wherein the immunoglobulin Fc region is a non-glycosylated Fc region.

31. The conjugate according to claim 21, wherein the immunoglobulin Fc region is selected from the group consisting of a heavy-chain constant region 1 (CH1) domain, a heavy-chain constant region 2 (CH2) domain, a heavy-chain constant region 3 (CH3) domain and a heavy-chain constant region 4 (CH4) domain;

a CH1 domain and a CH2 domain;

a CH1 domain and a CH3 domain;

a CH2 domain and a CH3 domain;

a combination of one or more domains and an immunoglobulin hinge region or a portion of the hinge region; and

a dimer of each domain of the heavy-chain constant regions and the light-chain constant region.

32. The conjugate according to claim 21, wherein the immunoglobulin Fc region comprises a modification of a native Fc region selected from the group consisting of:

(i) an amino acid modification to prevent formation of a disulfide bond;

(ii) elimination of one or more amino acid residues from the N-terminal end of the native Fc region;

(iii) addition of a methionine residue to the N-terminal end of the Fc region;

(iv) deletion of a complement-binding site; and

(v) deletion of an antibody dependent cell mediated cytotoxicity (ADCC) site.

33. The conjugate according to claim 21, wherein the immunoglobulin Fc region is an Fc region from an immunoglobulin selected from the group consisting of immunoglobulin G, immunoglobulin A, immunoglobulin D, immunoglobulin E, and immunoglobulin M.

34. The conjugate according to claim 33, wherein the immunoglobulin Fc region is an IgG4 Fc region.

35. The conjugate according to claim 21, wherein the immunoglobulin Fc region is a human IgG4-derived non-glycosylated Fc region.

36. The conjugate of claim 21, wherein the non-peptidyl polymer is linked via a covalent bond to the oxyntomodulin derivative at a cysteine residue.

37. A pharmaceutical composition comprising the conjugate of claim 21 and a pharmaceutically acceptable carrier.

38. The pharmaceutical composition according to claim 37, further comprising a glucagon-like peptide-1 receptor agonist, a leptin receptor agonist, a dipeptidyl peptidase-IV inhibitor, a Y5 receptor antagonist, a melanin-concentrating hormone receptor antagonist, a Y2/3 receptor agonist, a MC3/4 receptor agonist, a gastric/pancreatic lipase inhibitor, a 5HT2c agonist, a β

3A receptor agonist, an amylin receptor agonist, a ghrelin antagonist, or a ghrelin receptor antagonist.

39. A method for treating obesity, comprising administering the conjugate of claim 21 to a subject in need thereof.

40. The method of claim 39, which further comprises administering an additional drug selected from the group consisting of a glucagon-like peptide-1 receptor agonist, a leptin receptor agonist, a dipeptidyl peptidase-IV inhibitor, a Y5 receptor antagonist, a melanin-concentrating hormone receptor antagonist, a Y2/3 receptor agonist, a MC3/4 receptor agonist, a gastric/pancreatic lipase inhibitor, a 5HT2c agonist, a β

3A receptor agonist, an amylin receptor agonist, a ghrelin antagonist, and a ghrelin receptor antagonist.

41. A conjugate comprising:

an oxyntomodulin derivative comprising the amino acid sequence of SEQ ID NO;

26;

an immunoglobulin Fc region; and

a non-peptidyl polymer, wherein the non-peptidyl polymer links the oxyntomodulin derivative and the immunoglobulin Fc region via covalent bonds.

42. The conjugate according to claim 41, wherein the oxyntomodulin derivative activates glucagon-like peptide-1 receptor and glucagon receptor.

43. The conjugate according to claim 41, wherein the conjugate has anti-obesity effects.

44. The conjugate according to claim 41, wherein the non-peptidyl polymer is comprises polyethylene glycol, polypropylene glycol, copolymers of ethylene glycol and propylene glycol, polyoxyethylated polyols, polyvinyl alcohol, polysaccharides, polyvinyl ethyl ether, polylactic acid, polylacticglycolic acid, lipid polymers, hyaluronic acid, or combinations thereof.

45. The conjugate according to claim 44, wherein the non-peptidyl polymer is polyethylene glycol.

46. The conjugate according to claim 44, wherein the polysaccharide is dextran, chitin, or a combination thereof.

47. The conjugate according to claim 41, wherein one functional group selected from the group consisting of an amine group and a thiol group of the immunoglobulin Fc region is attached to one end of the non-peptidyl polymer, and one functional group selected from the group consisting of an amine group and a thiol group of the oxyntomodulin derivative is attached to the other end of the non-peptidyl polymer.

48. The conjugate according to claim 41, wherein the conjugate is prepared by covalently linking the oxyntomodulin derivative to the immunoglobulin Fc region via the non-peptidyl polymer, the non-peptidyl polymer having, prior to forming the conjugate, reactive functional groups at both ends.

49. The conjugate according to claim 48, wherein the reactive functional groups are an aldehyde group, a maleimide group, a succinimide derivative, or combinations thereof.

50. The conjugate according to claim 41, wherein the immunoglobulin Fc region is a non-glycosylated Fc region.

51. The conjugate according to claim 41, wherein the immunoglobulin Fc region is selected from the group consisting of a heavy-chain constant region 1 (CH1) domain, a heavy-chain constant region 2 (CH2) domain, a heavy-chain constant region 3 (CH3) domain and a heavy-chain constant region 4 (CH4) domain;

a CH1 domain and a CH2 domain;

a CH1 domain and a CH3 domain;

a CH2 domain and a CH3 domain;

a combination of one or more domains and an immunoglobulin hinge region or a portion of the hinge region; and

a dimer of each domain of the heavy-chain constant regions and the light-chain constant region.

52. The conjugate according to claim 41, wherein the immunoglobulin Fc region comprises a modification of a native Fc region selected from the group consisting of:

(i) an amino acid modification to prevent formation of a disulfide bond;

(ii) elimination of one or more amino acid residues from the N-terminal end of the native Fc region;

(iii) addition of a methionine residue to the N-terminal end of the Fc region;

(iv) deletion of a complement-binding site; and

(v) deletion of an antibody dependent cell mediated cytotoxicity (ADCC) site.

53. The conjugate according to claim 41, wherein the immunoglobulin Fc region is an Fc region from an immunoglobulin selected from the group consisting of immunoglobulin G, immunoglobulin A, immunoglobulin D, immunoglobulin E, and immunoglobulin M.

54. The conjugate according to claim 53, wherein the immunoglobulin Fc region is an IgG4 Fc region.

55. The conjugate according to claim 54, wherein the immunoglobulin Fc region is a human IgG4-derived non-glycosylated Fc region.

56. The conjugate of claim 41, wherein the non-peptidyl polymer is linked via a covalent bond to the oxyntomodulin derivative at a cysteine residue.

57. A pharmaceutical composition comprising the conjugate of claim 41 and a pharmaceutically acceptable carrier.

58. The pharmaceutical composition according to claim 57, further comprising a glucagon-like peptide-1 receptor agonist, a leptin receptor agonist, a dipeptidyl peptidase-IV inhibitor, a Y5 receptor antagonist, a melanin-concentrating hormone receptor antagonist, a Y2/3 receptor agonist, a MC3/4 receptor agonist, a gastric/pancreatic lipase inhibitor, a 5HT2c agonist, a β

3A receptor agonist, an amylin receptor agonist, a ghrelin antagonist, or a ghrelin receptor antagonist.

59. A method for treating obesity, comprising administering the conjugate of claim 41 to a subject in need thereof.

60. The method of claim 59, which further comprises administering an additional drug selected from the group consisting of a glucagon-like peptide-1 receptor agonist, a leptin receptor agonist, a dipeptidyl peptidase-IV inhibitor, a Y5 receptor antagonist, a melanin-concentrating hormone receptor antagonist, a Y2/3 receptor agonist, a MC3/4 receptor agonist, a gastric/pancreatic lipase inhibitor, a 5HT2c agonist, a β

3A receptor agonist, an amylin receptor agonist, a ghrelin antagonist, and a ghrelin receptor antagonist.

61. A conjugate comprising:

an oxyntomodulin derivative comprising the amino acid sequence of SEQ ID NO;

28;

an immunoglobulin Fc region; and

a non-peptidyl polymer selected from the group consisting of polyethylene glycol, polypropylene glycol, copolymers of ethylene glycol and propylene glycol, polyoxyethylated polyols, polyvinyl alcohol, polyvinyl ethyl ether, polylactic acid, polylacticglycolic acid, lipid polymers, hyaluronic acid, and combinations thereof, and wherein the non-peptidyl polymer links the oxyntomodulin derivative and the immunoglobulin Fc region via covalent bonds.

62. The conjugate according to claim 61, wherein the oxyntomodulin derivative activates glucagon-like peptide-1 receptor and glucagon receptor.

63. The conjugate according to claim 61, wherein the conjugate has anti-obesity effects.

64. The conjugate according to claim 61, wherein the non-peptidyl polymer is polyethylene glycol.

65. The conjugate according to claim 61, wherein one functional group selected from the group consisting of an amine group and a thiol group of the immunoglobulin Fc region is attached to one end of the non-peptidyl polymer, and one functional group selected from the group consisting of an amine group and a thiol group of the oxyntomodulin derivative is attached to the other end of the non-peptidyl.

66. The conjugate according to claim 61, wherein the conjugate is prepared by covalently linking the oxyntomodulin derivative to the immunoglobulin Fc region via the non-peptidyl polymer, the non-peptidyl polymer having, prior to forming the conjugate, reactive functional groups at both ends.

67. The conjugate according to claim 66, wherein the reactive functional groups are an aldehyde group, a maleimide group, a succinimide derivative, or combinations thereof.

68. The conjugate according to claim 61, wherein the immunoglobulin Fc region is a non-glycosylated Fc region.

69. The conjugate according to claim 61, wherein the immunoglobulin Fc region is selected from the group consisting of a heavy-chain constant region 1 (CH1) domain, a heavy-chain constant region 2 (CH2) domain, a heavy-chain constant region 3 (CH3) domain and a heavy-chain constant region 4 (CH4) domain;

a CH1 domain and a CH2 domain;

a CH1 domain and a CH3 domain;

a CH2 domain and a CH3 domain;

a combination of one or more domains and an immunoglobulin hinge region or a portion of the hinge region; and

a dimer of each domain of the heavy-chain constant regions and the light-chain constant region.

70. The conjugate according to claim 61, wherein the immunoglobulin Fc region comprises a modification of a native Fc region selected from the group consisting of:

(i) an amino acid modification to prevent formation of a disulfide bond;

(ii) elimination of one or more amino acid residues from the N-terminal end of the native Fc region;

(iii) addition of a methionine residue to the N-terminal end of the Fc region;

(iv) deletion of a complement-binding site; and

(v) deletion of an antibody dependent cell mediated cytotoxicity (ADCC) site.

71. The conjugate according to claim 61, wherein the immunoglobulin Fc region is an Fc region from an immunoglobulin selected from the group consisting of immunoglobulin G, immunoglobulin A, immunoglobulin D, immunoglobulin E, and immunoglobulin M.

72. The conjugate according to claim 71, wherein the immunoglobulin Fc region is an IgG4 Fc region.

73. The conjugate according to claim 61, wherein the immunoglobulin Fc region is a human IgG4-derived non-glycosylated Fc region.

74. The conjugate of claim 61, wherein the non-peptidyl polymer is linked via a covalent bond to the oxyntomodulin derivative at a cysteine residue.

75. A pharmaceutical composition comprising the conjugate of claim 61 and a pharmaceutically acceptable carrier.

76. The pharmaceutical composition according to claim 75, further comprising a glucagon-like peptide-1 receptor agonist, a leptin receptor agonist, a dipeptidyl peptidase-IV inhibitor, a Y5 receptor antagonist, a melanin-concentrating hormone receptor antagonist, a Y2/3 receptor agonist, a MC3/4 receptor agonist, a gastric/pancreatic lipase inhibitor, a 5HT2c agonist, a β

3A receptor agonist, an amylin receptor agonist, a ghrelin antagonist, or a ghrelin receptor antagonist.

77. A method for treating obesity, comprising administering the conjugate of claim 61 to a subject in need thereof.

78. The method of claim 77, which further comprises administering an additional drug selected from the group consisting of a glucagon-like peptide-1 receptor agonist, a leptin receptor agonist, a dipeptidyl peptidase-IV inhibitor, a Y5 receptor antagonist, a melanin-concentrating hormone receptor antagonist, a Y2/3 receptor agonist, a MC3/4 receptor agonist, a gastric/pancreatic lipase inhibitor, a 5HT2c agonist, a β

3A receptor agonist, an amylin receptor agonist, a ghrelin antagonist, and a ghrelin receptor antagonist.