G-type peptides to ameliorate atherosclerosis

• US 6,930,085 B2
• Filed: 04/05/2002
• Issued: 08/16/2005
• Est. Priority Date: 04/05/2002
• Status: Expired due to Term

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Fogelman, Alan M., Navab, Mohamad
• Primary Examiner(s): RUSSEL, JEFFREY E
• Application Number:
  US10/120,508
• Publication Number:
  US 20030191057A1
• Time in Patent Office: 1,229 Days
• Field of Search: 530/300, 530/324, 530/325, 530/326, 530/327, 530/328, 530/345, 436/86, 436/87, 514/2, 514/7, 514/12, 514/13, 514/14, 514/15
• US Class Current: 424/94.4
• 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: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 33/00 Antiparasitic agents
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/775 Apolipopeptides

Abstract

This invention provides novel peptides that ameliorate one or more symptoms of atherosclerosis and/or other pathologies characterized by an inflammatory response. In certain embodiment, the peptides resemble a G* amphipathic helix of apolipoprotein J. The peptides are highly stable and readily administered via an oral route.

Claims

1. An isolated polypeptide that ameliorates a symptom of atherosclerosis or other pathology associated with an inflammatory response, said polypeptide comprising an amphipathic helical peptide having charged residues on the polar face of the peptide and possessing a wide non-polar face, wherein said peptide comprises an amino acid sequence selected from the group consisting of LLEQLNEQFNWVSRLANL (SEQ ID NO:

2), and LVGRQLEEFL, (SEQ ID NO;

9), and wherein said polypeptide is 40 or fewer amino acids in length and at least 10 amino acids in length.

2. The polypeptide of claim 1, wherein said peptide comprises a G* amphipathic helix.

3. The polypeptide of claim 2, wherein said peptide shows greater than about 50% sequence identity with apo J.

4. The polypeptide of claim 1, wherein said peptide protects a phospholipid against oxidation by an oxidizing agent.

5. The polypeptide of claim 1, wherein said peptide is a concatamer of two or more of said amino acid sequences.

6. The polypeptide of claim 1, wherein said peptide further comprises a protecting group.

7. The polypeptide of claim 1, wherein said peptide further comprises a protecting group coupled to the amino or carboxyl terminus.

8. The polypeptide of claim 6, wherein said protecting group is a protecting group selected from the group consisting of an acetyl, amide, 3 to 20 carbon alkyl groups, Fmoc, 9-fluoreneacetyl group, 1-fluorenecarboxylic group, 9-fluorenecarboxylic, 9-fluorenone-1-carboxylic group, benzyloxycarbonyl, Xanthyl (Xan), Trityl (Trt), 4-methyltrityl (Mtt), 4-methoxytrityl (Mint), 4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr), Mesitylene-2-sulphonyl (Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl (Tos), 2,2,5,7,8-pentamethyl chroman-6-sulphonyl (Pmc), 4-methylbenzyl (MeBzl), 4-methoxybenzyl (MeOBzl), Benzyloxy (BzlO), Benzyl (Bzl), Benzoyl (Bz), 3-nitro-2-pyridinesulphenyl (Npys), 1-(4,4dimethyl-2,6-dioxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl (2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl (2-Cl-Z), 2-bromobenzyloxycarbonyl (2-Br-Z), Benzyloxymethyl (Bom), t-butoxycarbonyl (Boc), cyclohexyloxy (cHxO),t-butoxymethyl (Bum), t-butoxy (tBuO), t-Butyl (tBu), a propyl group, a butyl group, a pentyl group, a hexyl group, and Trifluoroacetyl (TFA).

9. The polypeptide of claim 6, wherein said peptide comprises a protecting group coupled to the amino terminus and said amino terminal protecting group is a protecting group selected from the group consisting of a benzoyl group, an acetyl, a propionyl, a carbobenzoxy, a propyl, a butyl, a pentyl, a hexyl, and a 3 to 20 carbon alkyl.

10. The polypeptide of claim 6, wherein said peptide comprises a protecting group coupled to the carboxyl terminus and said carboxyl terminal protecting group is an amide.

11. The polypeptide of claim 6, wherein said peptide further comprises:

a first protecting group coupled to the amino terminus wherein said protecting group is a protecting group selected from the group consisting of a benzoyl group, an acetyl, a propionyl, a carbobenzoxy, a propyl, a butyl, a pentyl, a hexyl, and a 3 to 20 carbon alkyl; and

a second protecting group coupled to the carboxyl terminus and said carboxyl terminal protecting group is an amide.

12. The polypeptide of claim 1, wherein said peptide comprises a first protecting group coupled to the amino terminus and a second protecting group coupled to the carboxyl terminus.

13. The polypeptide of claim 1, wherein said polypeptide comprises an Ac group on the amino terminus and an —

NH₂ on the carboxyl terminus.

14. The polypeptide of claim 1, wherein said peptide comprises a “

D”

amino acid.

15. The polypeptide of claim 1, wherein said peptide comprises a plurality of “

D”

amino acids.

16. The polypeptide of claim 1, wherein all enantiomeric amino acids comprising said peptide are “

D”

amino acids.

17. The polypeptide of claim 1, wherein said polypeptide is mixed with a pharmacologically acceptable excipient.

18. The polypeptide of claim 1, wherein said polypeptide is mixed with a pharmacologically acceptable excipient suitable for oral administration to a mammal.

39. A pharmaceutical composition, said composition comprising a polypeptide of any of claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 in a pharmaceutically acceptable excipient.

40. The composition of claim 39, wherein said composition is in the form of a unit dosage formulation.

63. A method of ameliorating a symptom of atherosclerosis in a mammal, said method comprising administering to said mammal a polypeptide according to any of claims 1, 2, 3, 4, 5-12, 13, 16, 19-23, and 4562.

64. The method of claim 63, wherein said administering comprises orally administering said polypeptide.

65. The method of claim 63, wherein said mammal is a mammal diagnosed as having one or more symptoms of atherosclerosis.

66. The method of claim 63, wherein said mammal is a mammal diagnosed as at risk for atherosclerosis.

67. The method of claim 63, wherein said mammal is a human.

68. The method of claim 63, wherein said mammal is non-human mammal.

69. A method of ameliorating a symptom of a pathology characterized by an inflammatory response in a mammal, said method comprising administering to said mammal a polypeptide according to any of claims 1, 2, 3, 4, 5-12, 13-16, 19-23, and 45-62.

70. The method of claim 69, wherein said mammal is a mammal diagnosed as having one or more symptoms of an inflammatory response.

71. The method of claim 69, wherein said mammal is a mammal diagnosed as at risk for a pathology associated with an inflammatory response.

72. The method of claim 69, wherein said mammal is a human.

73. The method of claim 69, wherein said mammal is a non-human mammal.

74. A kit for ameliorating a symptom of atherosclerosis, said kit comprising a container containing a polypeptide according to any of claims 2, 3, 4, 5-12, 13-18, 19-23, and 45-62.

75. The kit of claim 74, wherein said polypeptide is combined with a pharmaceutically acceptable excipient in a unit dosage formulation.

76. The kit of claim 75, wherein said unit dosage formulation is for oral administration.

77. The kit of claim 74, further comprising instructional materials teaching the use of said polypeptide or said composition for ameliorating one or more symptoms of atherosclerosis or of a pathology characterized by an inflammatory response.

78. A method of mitigating or preventing a coronary complication associated with an acute phase response to an inflammation in a mammal, wherein said coronary complication is a symptom of atherosclerosis, said method comprising administering to a mammal having said acute phase response or at risk for said acute phase response, a polypeptide according to any of claims 1, 2, 3, 4, 5-12, 13-16, 19-23, and 45-62.

79. The method of claim 78, said administration is by a route selected from the group consisting of oral administration, nasal administration, rectal administration, intraperitoneal injection, intravascular injection, subcutaneous injection, transcutaneous administration, and intramuscular injection.

80. The method of claim 78, wherein said polypeptide is administered in combination with an all L-form of the same polypeptide.

81. The method of claim 78, wherein said polypeptide is provided as a unit formulation in a pharmaceutically acceptable excipient.

82. The method of claim 78, wherein said acute phase response is an inflammatory response associated with a recurrent inflammatory disease.

83. The method of claim 79, wherein said acute phase response is associated with a disease selected from the group consisting of leprosy, tuberculosis, systemic lupus erythematosus, polymyalgia rheumatica, polyarteritis nodosa, scleroderma, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, coronary calcification, calcific aortic stenosis, osteoporosis, and rheumatoid arthritis.

84. The method of claim 78, wherein said acute phase response is an inflammatory response associated with a condition selected from the group consisting of a bacterial infection, a viral infection, a fungal infection, an organ transplant, a wound, an implanted prosthesis, parasitic infection, sepsis, endotoxic shock syndrome, and biofilm formation.

85. A method of mitigating or preventing a coronary complication associated with an acute phase response to an inflammation in a mammal, wherein said coronary complication is a symptom of atherosclerosis, said method comprising:

assaying said mammal for an acute phase protein (APP) level indicative of an acute phase response or a significant risk of an acute phase response; and

administering to a mammal showing an acute phase protein (APP) level indicative of an acute phase response a polypeptide according to any of claims 1, 2, 3, 4, 5-12, 13-16, 19-23, and 45-62.

86. The method of claim 85, wherein said acute phase protein (APP) is a postive APR selected from the group consisting of serum amyloid A, c-reactive protein, serum amyloid P component, C2 complement protein, C3 complement protein, C4 complement protein, C5 complement protein, C9 complement protein, B complement protein, C1 inhibitor, C4 binding protein, fibrinogen, von Willebrand factor, α

1-antitrypsin, α

1-antichymotrypsin, α

2 antiplasmin, heparin cofactor II, plasminogen activator inhibitor I, haptoglobin, haemopexin, ceruloplasmin , manganese superoxide dismutase, α

1-acid glycoprotein, haeme oxygenase, mannose binding protein, leukocyte protein I, lipoprotein (a), and lipopolysaccharide binding protein.

87. The method of claim 85, wherein said acute phase protein (APP) is a negative APR selected from the group consisting of albumin, prealbumin, transferin, apoAI, apoAII, α

2-HS glycoprotein, inter-α

-trypsin inhibitor, and histidine-rich glycoprotein.

88. The method of claim 63, wherein said polypeptide is in a pharmaceutically acceptable excipient.

89. The method of claim 63, wherein said polypeptide is in a pharmaceutically acceptable excipient suitable for oral administration to a mammal.

90. The method of claim 63, wherein said polypeptide is in a pharmaceutically acceptable excipient in the form of a unit dosage formulation.

91. The method of claim 69, wherein said polypeptide is in a pharmaceutically acceptable excipient.

92. The method of claim 69, wherein said polypeptide is in a pharmaceutically acceptable excipient suitable for oral administration to a mammal.

93. The method of claim 69, wherein said polypeptide is in a pharmaceutically acceptable excipient in the form of a unit dosage formulation.

94. The kit of claim 74, wherein said polypeptide is in a pharmaceutically acceptable excipient.

95. The kit of claim 74, wherein said polypeptide is in a pharmaceutically acceptable excipient suitable for oral administration to a mammal.

96. The kit method of claim 74, wherein said polypeptide is in a pharmaceutically acceptable excipient in the form of a unit dosage formulation.

97. The method of claim 78, wherein said polypeptide is in a pharmaceutically acceptable excipient.

98. The method of claim 78, wherein said polypeptide is in a pharmaceutically acceptable excipient suitable for oral administration to a mammal.

99. The method of claim 78, wherein said polypeptide is in a pharmaceutically acceptable excipient in the form of a unit dosage formulation.

100. The method of claim 85, wherein said polypeptide is in a pharmaceutically acceptable excipient.

101. The method of claim 85, wherein said polypeptide is in a pharmaceutically acceptable excipient suitable for oral administration to a mammal.

102. The method of claim 85, wherein said polypeptide is in a pharmaceutically acceptable excipient in the form of a unit dosage formulation.

19. An isolated polypeptide that ameliorates a symptom of atherosclerosis or other pathology associated with an inflammatory response, said polypeptide comprising an amphipathic helical peptide having charged residues on the polar face of the peptide and possessing a wide non-polar face, wherein said polypeptide is coupled to a phospholipid.

20. The polypeptide of claim 19, wherein said polypeptide is covalently coupled to a phospholipid.

21. The polypeptide of claim 19, wherein said polypeptide is covalently coupled to a phospholipid comprising lysophosphatidyl choline.

22. The polypeptide of claim 19, wherein said polypeptide is covalently coupled to a phospholipid comprising a fatty acid selected from the group consisting of propionoyl, butanoyl, pentanoyl, caproyl, heptanoyl, capryloyl, nonanoyl, capryl, undecanoyl, lauroyl, tridecanoyl, myristoyl, pentadecanoyl, palmitoyl, heptadecanoyl, stearoyl, nonadecanoyl, arachidoyl, heniecosanoyl, behenoyl, trucisanoyl, lignoceroyl, myristoleoyl (9-cis), myristelaidoyl (9-trans), palmitoleoyl (9-cis), and palmitelaidoyl (9-trans).

23. The polypeptide of claim 22, wherein said polypeptide is covalently coupled to the sn-1 or sn-2 position of said phospholipid.

45. The polypeptide of claim 19, wherein said peptide is at least 10 amino acids in length.

46. The polypeptide of claim 19, wherein said peptide is about 40 or fewer amino acids in length.

47. The polypeptide of claim 19, wherein said peptide comprises a G* amphipathic helix.

48. The polypeptide of claim 47, wherein said peptide shows greater than about 50% sequence identity with apo J.

49. The polypeptide of claim 19, wherein said peptide comprises an amino acid sequence selected from the group consisting of LLEQLNEQFNWVSRLANL (SEQ ID NO:

2), and LVGRQLEEFL (SEQ ID NO;

9).

50. The polypeptide of claim 49, wherein said peptide is a concatamer of two or more of said amino acid sequences.

51. The polypeptide of claim 19, wherein said peptide further comprises a protecting group coupled to the amino or carboxyl terminus.

52. The polypeptide of claim 51, wherein said protecting group is a protecting group selected from the group consisting of an acetyl, amide, 3 to 20 carbon alkyl groups, Fmoc, 9-fluoreneacetyl group, 1-fluorenecarboxylic group, 9-fluorenecarboxylic, 9-fluorenone-1-carboxylic group, benzyloxycarbonyl, Xanthyl (Xan), Trityl (Trt), 4-methyltrityl (Mtt), 4-methoxytrityl (Mint), 4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr), Mesitylene-2-sulphonyl (Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl (Tos), 2,2,5,7,8-pentamethyl chroman-6-sulphonyl (Pmc), 4-methylbenzyl (MeBzl), 4-methoxybenzyl (MeOBzl), Benzyloxy (BzlO), Benzyl (Bzl), Benzoyl (Bz), 3-nitro-2-pyridinesulphenyl (Npys), 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl (2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl (2-Cl-Z), 2-bromobenzyloxycarbonyl (2-Br-Z), Benzyloxymethyl (Bom), t-butoxycarbonyl (Boc), cyclohexyloxy (cHxO), t-butoxymethyl (Bum), t-butoxy (tBuO), t-Butyl (tBu), a propyl group, a butyl group, a pentyl group, a hexyl group, and Trifluoroacetyl (TFA).

53. The polypeptide of claim 51, wherein said peptide comprises a protecting group coupled to the amino terminus and said amino terminal protecting group is a protecting group selected from the group consisting of a benzoyl group, an acetyl, a propionyl, a carbobenzoxy, a propyl, a butyl, a pentyl, a hexyl, and a 3 to 20 carbon alkyl.

54. The polypeptide of claim 51, wherein said peptide comprises a protecting group coupled to the carboxyl terminus and said carboxyl terminal protecting group is an amide.

55. The polypeptide of claim 51, wherein said peptide further comprises:

a first protecting group coupled to the amino terminus wherein said protecting group is a protecting group selected from the group consisting of a benzoyl group, an acetyl, a propionyl, a carbobenzoxy, a propyl, a butyl, a pentyl, a hexyl, and a 3 to 20 carbon alkyl; and

a second protecting group coupled to the carboxyl terminus and said carboxyl terminal protecting group is an amide.

56. The polypeptide of claim 51, wherein said peptide comprises a first protecting group coupled to the amino terminus and a second protecting group coupled to the carboxyl terminus.

57. The polypeptide of claim 51, wherein said peptide comprises an Ac group on the amino terminus.

58. The polypeptide of claim 51, wherein said peptide comprises an —

NH₂ on the carboxyl terminus.

59. The polypeptide of claim 51, wherein said peptide comprises an Ac group on the amino terminus and an —

NH₂ on the carboxyl terminus.

60. The polypeptide of claim 51, wherein said peptide comprises a “

D”

amino acid.

61. The polypeptide of claim 51, wherein said peptide comprises a plurality of “

D”

amino acids.

62. The polypeptide of claim 51, wherein all enantiomeric amino acids comprising said peptide are “

D”

amino acids.

24. A composition suitable for oral administration that ameliorates a symptom of atherosclerosis, wherein said composition comprises a peptide comprising an amphipathic helix having charged residues on the polar face of the peptide and possessing a wide non-polar face, wherein said peptide comprises a D amino acid and said peptide is blocked at the amino terminus and the carboxyl terminus.

25. The composition of claim 24, wherein said peptide is at least 10 amino acids in length.

26. The composition of claim 25, wherein said peptide is about 40 or fewer peptides in length.

27. The composition of claim 25, wherein said peptide comprises a G* amphipathic helix.

28. The composition of claim 27, wherein said peptide shows greater than about 50% sequence identity with apo J.

29. The composition of claim 25, wherein said peptide protects a phospholipid against oxidation by an oxidizing agent.

30. The composition of claim 25, wherein said peptide comprises an amino acid sequence selected from the group consisting of LLEQLNEQFNWVSRLANL (SEQ ID NO:

2), and LVGRQLEEFL (SEQ ID NO;

9).

31. The composition of claim 25, wherein said peptide is blocked at the amino terminus with a first protecting group and said peptide is blocked at the carboxyl terminus with a second protecting group and said first protecting group and said second protecting group are independently selected from the group consisting of an acetyl, amide, 3 to 20 carbon alkyl groups, Fmoc, 9-fluoreneacetyl group, 1-fluorenecarboxylic group, 9-fluorenecarboxylic, 9-fluorenone-1-carboxylic group, benzyloxycarbonyl, Xanthyl (Xan), Trityl (Trt), 4-methyltrityl (Mtt), 4-methoxytrityl (Mmt), 4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr), Mesitylene-2-sulphonyl (Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl (Tos), 2,2,5,7,8-pentamethyl chroman-6-sulphonyl (Pmc), 4-methylbenzyl (MeBzl), 4-methoxyberizyl (MeOBzl), Benzyloxy (BzlO), Benzyl (Bzl), Benzoyl (Bz), 3-nitro-2-pyridinesulphenyl (Npys), 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl (2,6-DiCI-Bzl), 2-chlorobenzyloxycarbonyl (2-Cl-Z), 2-bromobenzyloxycarbonyl (2-Br-Z), Benzyloxymethyl (Bom), t-butoxycarbonyl (Boc), cyclohexyloxy (cHxO), t-butoxymethyl (Bum), t-butoxy (tBuO), t-Butyl (tBu), a propyl group, a butyl group, a pentyl group, a hexyl group, and Trifluoroacetyl (TFA).

32. The composition of claim 25, wherein said first protecting group is an acetyl.

33. The composition of claim 25, wherein said second protecting group is an amide.

34. The composition of claim 25, wherein more than half of the enantiomeric amino acids comprising said peptide are D amino acids.

35. The composition of claim 25, wherein all enantiomeric amino acids comprising said peptide are D amino acids.

36. The composition of claim 25, wherein said composition further comprises a pharmaceutically acceptable excipient.

37. The composition of claim 36, wherein said excipient is an excipient suitable for oral administration.

38. The composition of claim 36, wherein said excipient is an excipient suitable for injection.

41. The composition of claim 25, wherein said peptide is coupled to a phospholipid.

42. The composition of claim 41, wherein said peptide is covalently coupled to a phospholipid.

43. The composition of claim 41, wherein said peptide is covalently coupled to a phospholipid comprising lysophosphatidyl choline.

44. The composition of claim 41, wherein said peptide is covalently coupled to a phospholipid comprising a fatty acid selected from the group consisting of propionoyl, butanoyl, pentanoyl, caproyl, heptanoyl, capryloyl, nonanoyl, capryl, undecanoyl, lauroyl, tridecanoyl, myristoyl, pentadecanoyl, palmitoyl, heptadecanoyl, stearoyl, nonadecanoyl, arachidoyl, heniecosanoyl, behenoyl, trucisanoyl, lignoceroyl, myristoleoyl (9-cis), myristelaidoyl (9-trans), palmitoleoyl (9-cis), and palmitelaidoyl (9-trans).