Compositions and methods for modulating angiopoietin-like 3 expression

• US 9,957,292 B2
• Filed: 12/04/2015
• Issued: 05/01/2018
• Est. Priority Date: 05/01/2014
• Status: Active Grant

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: Ionis Pharmaceuticals, Inc.
• Inventors: Prakash, Thazha P., Seth, Punit P., Swayze, Eric E., Freier, Susan M., Graham, Mark J., Crooke, Rosanne M.
• Primary Examiner(s): McIntosh, III, Traviss C
• Application Number:
  US14/959,714
• Publication Number:
  US 20160194349A1
• Time in Patent Office: 879 Days
• Field of Search: None
• 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: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• 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: A61P 9/12 Antihypertensives
• CPC Class Codes: C07H 15/04 attached to an oxygen atom of the saccharide radical
• CPC Class Codes: C07H 19/20 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C07H 21/02 with ribosyl as saccharide radical
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C12N 15/1136 against growth factors, growth regulators, cytokines, lymphokines or hormones
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3231 having an additional ring, e.g. LNA, ENA
• CPC Class Codes: C12N 2310/3341 5-Methylcytosine
• CPC Class Codes: C12N 2310/341 Gapmers, i.e. of the type ===---===
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/351 Conjugate
• CPC Class Codes: C12N 2310/3525 : MOE, methoxyethoxy
• CPC Class Codes: C12N 2320/32 : Special delivery means, e.g. tissue-specific

Abstract

Provided herein are methods, compounds, and compositions for reducing expression of an ANGPTL3 mRNA and protein in an animal. Also provided herein are methods, compounds, and compositions for reducing lipids and/or glucose in an animal. Such methods, compounds, and compositions are useful to treat, prevent, delay, or ameliorate any one or more of cardiovascular disease and/or metabolic disease, or a symptom thereof, in an individual in need thereof.

Claims

1. A compound comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides having a nucleobase sequence comprising at least 12 contiguous nucleobases complementary to an equal length portion of any of SEQ ID NOs:

1-2, and wherein the conjugate group comprises;

2. The compound of claim 1, wherein the modified oligonucleotide comprises at least one modified sugar.

3. The compound of claim 2, wherein at least one modified sugar is a bicyclic sugar.

4. The compound of claim 2, wherein at least one modified sugar comprises a 2′

-O-methoxyethyl, a constrained ethyl, a 3′

-fluoro-HNA or a 4′

-(CH₂)_n-O-2′

bridge, wherein n is 1 or 2.

5. The compound of claim 2, wherein at least one modified sugar is 2′

-O-methoxyethyl.

6. The compound of claim 1, wherein the modified oligonucleotide comprises at least one modified nucleobase.

7. The compound of claim 6, wherein the modified nucleobase is a 5-methylcytosine.

8. The compound of claim 1, wherein the conjugate group is linked to the modified oligonucleotide at the 5′

end of the modified oligonucleotide.

9. The compound of claim 1, wherein the conjugate group is linked to the modified oligonucleotide at the 3′

end of the modified oligonucleotide.

10. The compound of claim 1, wherein each internucleoside linkage of the modified oligonucleotide is selected from a phosphodiester internucleoside linkage and a phosphorothioate internucleoside linkage.

11. The compound of claim 10, wherein the modified oligonucleotide comprises at least 5 phosphodiester internucleoside linkages.

12. The compound of claim 10, wherein the modified oligonucleotide comprises at least 2 phosphorothioate internucleoside linkages.

13. The compound of claim 1, wherein the modified oligonucleotide is single stranded.

14. The compound of claim 1, wherein the modified oligonucleotide is double stranded.

15. The compound of claim 1, wherein the modified oligonucleotide comprises:

a gap segment consisting of linked deoxynucleosides;

a 5′

wing segment consisting of linked nucleosides;

a 3′

wing segment consisting of linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

16. The compound of claim 15, wherein each internucleoside linkage in the gap segment of the modified oligonucleotide is a phosphorothioate linkage.

17. The compound of claim 16, wherein the modified oligonucleotide further comprises at least one phosphorothioate internucleoside linkage in each wing segment.

18. The compound of claim 1, wherein the modified oligonucleotide is 20 linked nucleosides in length and comprises:

a gap segment consisting of ten linked deoxynucleosides;

a 5′

wing segment consisting of five linked nucleosides;

a 3′

wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment, wherein each nucleoside of each wing segment comprises a 2′

-O-methoxyethyl sugar, and wherein each cytosine residue is a 5-methylcytosine.

19. The compound of claim 18, wherein each internucleoside linkage in the gap segment of the modified oligonucleotide is a phosphorothioate linkage.

20. The compound of claim 19, wherein the modified oligonucleotide further comprises at least one phosphorothioate internucleoside linkage in each wing segment.

21. The compound of claim 1, wherein the modified oligonucleotide consists of 20 linked nucleosides.

22. The compound of claim 1, wherein the modified oligonucleotide comprises the formula

G_esG_eoA_eo^mC_eoA_eoT_dsT_dsG_ds^mC_ds^mC_dsA_dsG_dsT_dsA_dsA_dsT_eo^mC_eoG_es^mC_esA_e,

wherein “

e”

indicates a 2′

-O-methoxyethyl modified nucleoside, “

d”

indicates a β

-D-2′

-deoxyribonucleoside, “

s”

indicates a phosphorothioate internucleoside linkage, “

o”

indicates a phosphodiester internucleoside linkage, and “

m”

indicates a 5-methylcytosine.

23. A composition comprising the compound of claim 1, or a salt thereof, and a pharmaceutically acceptable carrier or diluent.

24. A composition comprising the compound of claim 22, or a salt thereof, and a pharmaceutically acceptable carrier or diluent.

25. A method for reducing angiopoietin-like 3 (ANGPTL3) expression in a subject, comprising administering to the subject a compound, or salt thereof, comprising a modified oligonucleotide and a conjugate group, wherein:

the modified oligonucleotide consists of 12 to 30 linked nucleosides having a nucleobase sequence comprising at least 12 contiguous nucleobases complementary to an equal length portion of any of SEQ ID NOs;

1-2;

wherein the conjugate group comprises;

27. The method of claim 25, wherein the modified oligonucleotide comprises at least one modified sugar.

28. The method of claim 27, wherein at least one modified sugar is a bicyclic sugar.

29. The method of claim 27, wherein at least one modified sugar comprises a 2′

-O-methoxyethyl, a constrained ethyl, a 3′

-fluoro-HNA or a 4′

-(CH₂)_n-O-2′

bridge, wherein n is 1 or 2.

30. The method of claim 27, wherein at least one modified sugar is 2′

-O-methoxyethyl.

31. The method of claim 25, wherein the modified oligonucleotide comprises at least one modified nucleobase.

32. The method of claim 31, wherein the modified nucleobase is a 5-methylcytosine.

33. The method of claim 25, wherein the conjugate group is linked to the modified oligonucleotide at the 5′

end of the modified oligonucleotide.

34. The method of claim 25, wherein the conjugate group is linked to the modified oligonucleotide at the 3′

end of the modified oligonucleotide.

35. The method of claim 25, wherein each internucleoside linkage of the modified oligonucleotide is selected from a phosphodiester internucleoside linkage and a phosphorothioate internucleoside linkage.

36. The method of claim 35, wherein the modified oligonucleotide comprises at least 5 phosphodiester internucleoside linkages.

37. The method of claim 35, wherein the modified oligonucleotide comprises at least 2 phosphorothioate internucleoside linkages.

38. The method of claim 25, wherein the modified oligonucleotide is single stranded.

39. The method of claim 25, wherein the modified oligonucleotide is double stranded.

40. The method of claim 25, wherein the modified oligonucleotide comprises:

a gap segment consisting of linked deoxynucleosides;

a 5′

wing segment consisting of linked nucleosides;

a 3′

wing segment consisting of linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

41. The method of claim 40, wherein each internucleoside linkage in the gap segment of the modified oligonucleotide is a phosphorothioate linkage.

42. The method of claim 41, wherein the modified oligonucleotide further comprises at least one phosphorothioate internucleoside linkage in each wing segment.

43. The method of claim 25, wherein the modified oligonucleotide is 20 linked nucleosides in length and comprises:

a gap segment consisting of ten linked deoxynucleosides;

a 5′

wing segment consisting of five linked nucleosides;

a 3′

wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment, wherein each nucleoside of each wing segment comprises a 2′

-O-methoxyethyl sugar, and wherein each cytosine residue is a 5-methylcytosine.

44. The method of claim 43, wherein each internucleoside linkage in the gap segment of the modified oligonucleotide is a phosphorothioate linkage.

45. The method of claim 44, wherein the modified oligonucleotide further comprises at least one phosphorothioate internucleoside linkage in each wing segment.

46. The method of claim 25, wherein the modified oligonucleotide consists of 20 linked nucleosides.

47. The method of claim 25, wherein the modified oligonucleotide comprises the formula G_esG_eoA_eo^mC_eoA_eoT_dsT_dsG_ds^mC_ds^mC_dsA_dsG_dsT_dsA_dsA_dsT_eo^mC_eoG_es^mC_esA_e, wherein “

e”

indicates a 2′

-O-methoxyethyl modified nucleoside, “

d”

indicates a β

-D-2′

-deoxyribonucleoside, “

s”

indicates a phosphorothioate internucleoside linkage, “

o”

indicates a phosphodiester internucleoside linkage, and “

m”

indicates a 5-methylcytosine.

48. The method of claim 25, wherein a composition comprising the compound, or salt thereof, is administered to the subject, and the composition comprises a pharmaceutically acceptable carrier or diluent.

49. The method of claim 47, wherein a composition comprising the compound, or salt thereof, is administered to the subject, and the composition comprises a pharmaceutically acceptable carrier or diluent.

50. The method of claim 25, wherein the subject is human.

26. A method for treating, delaying, slowing the progression of and/or ameliorating a cardiovascular and/or metabolic disease, disorder or condition, comprising administering to a subject in need thereof an effective amount of a compound, or salt thereof, comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides having a nucleobase sequence comprising at least 12 contiguous nucleobases complementary to an equal length portion of any of SEQ ID NOs:

1-2, and wherein the conjugate group comprises;

51. The method of claim 26, wherein the modified oligonucleotide comprises at least one modified sugar.

52. The method of claim 51, wherein at least one modified sugar is a bicyclic sugar.

53. The method of claim 51, wherein at least one modified sugar comprises a 2′

-O-methoxyethyl, a constrained ethyl, a 3′

-fluoro-HNA or a 4′

-(CH₂)_n-O-2′

bridge, wherein n is 1 or 2.

54. The method of claim 51, wherein at least one modified sugar is 2′

-O-methoxyethyl.

55. The method of claim 26, wherein the modified oligonucleotide comprises at least one modified nucleobase.

56. The method of claim 55, wherein the modified nucleobase is a 5-methylcytosine.

57. The method of claim 26, wherein the conjugate group is linked to the modified oligonucleotide at the 5′

end of the modified oligonucleotide.

58. The method of claim 26, wherein the conjugate group is linked to the modified oligonucleotide at the 3′

end of the modified oligonucleotide.

59. The method of claim 26, wherein each internucleoside linkage of the modified oligonucleotide is selected from a phosphodiester internucleoside linkage and a phosphorothioate internucleoside linkage.

60. The method of claim 59, wherein the modified oligonucleotide comprises at least 5 phosphodiester internucleoside linkages.

61. The method of claim 59, wherein the modified oligonucleotide comprises at least 2 phosphorothioate internucleoside linkages.

62. The method of claim 26, wherein the modified oligonucleotide is single stranded.

63. The method of claim 26, wherein the modified oligonucleotide is double stranded.

64. The method of claim 26, wherein the modified oligonucleotide comprises:

a gap segment consisting of linked deoxynucleosides;

a 5′

wing segment consisting of linked nucleosides;

a 3′

wing segment consisting of linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

65. The method of claim 64, wherein each internucleoside linkage in the gap segment of the modified oligonucleotide is a phosphorothioate linkage.

66. The method of claim 65, wherein the modified oligonucleotide further comprises at least one phosphorothioate internucleoside linkage in each wing segment.

67. The method of claim 26, wherein the modified oligonucleotide is 20 linked nucleosides in length and comprises:

a gap segment consisting of ten linked deoxynucleosides;

a 5′

wing segment consisting of five linked nucleosides;

a 3′

wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment, wherein each nucleoside of each wing segment comprises a 2′

-O-methoxyethyl sugar, and wherein each cytosine residue is a 5-methylcytosine.

68. The method of claim 67, wherein each internucleoside linkage in the gap segment of the modified oligonucleotide is a phosphorothioate linkage.

69. The method of claim 68, wherein the modified oligonucleotide further comprises at least one phosphorothioate internucleoside linkage in each wing segment.

70. The method of claim 26, wherein the modified oligonucleotide consists of 20 linked nucleosides.

71. The method of claim 26, wherein the modified oligonucleotide comprises the formula G_esG_eoA_eo^mC_eoA_eoT_dsT_dsG_ds^mC_ds^mC_dsA_dsG_dsT_dsA_dsA_dsT_eo^mC_eoG_es^mC_esA_e, wherein “

e”

indicates a 2′

-O-methoxyethyl modified nucleoside, “

d”

indicates a β

-D-2′

-deoxyribonucleoside, “

s”

indicates a phosphorothioate internucleoside linkage, “

o”

indicates a phosphodiester internucleoside linkage, and “

m”

indicates a 5-methylcytosine.

72. The method of claim 26, wherein a composition comprising the compound, or salt thereof, is administered to the subject, and the composition comprises a pharmaceutically acceptable carrier or diluent.

73. The method of claim 71, wherein a composition comprising the compound, or salt thereof, is administered to the subject, and the composition comprises a pharmaceutically acceptable carrier or diluent.

74. The method of claim 26, wherein the subject is human.

75. The method of claim 26, wherein the disease, disorder or condition is atherosclerosis, hepatic steatosis, obesity, diabetes, dyslipidemia, coronary heart disease, non-alcoholic fatty liver disease (NAFLD), hyperfattyacidemia and/or metabolic syndrome.

76. The method of claim 75, wherein the hepatic steatosis is nonalcoholic steatohepatitis (NASH) and the dyslipidemia is hyperlipidemia, hypercholesterolemia and/or hypertriglyceridemia.

77. The method of claim 75, wherein the disease, disorder or condition is non-alcoholic fatty liver disease (NAFLD).

78. The method of claim 75, wherein the disease, disorder or condition is hepatic steatosis and the hepatic steatosis is nonalcoholic steatohepatitis (NASH).

79. The method of claim 75, wherein the disease, disorder or condition is dyslipidemia and the dyslipidemia is hypercholesterolemia.

80. The method of claim 26, wherein the administering is by parenteral administration.

81. The method of claim 80, wherein the administering is by injection.