Modulation of apolipoprotein CIII (ApoCIII) expression

• US 9,157,082 B2
• Filed: 04/27/2012
• Issued: 10/13/2015
• Est. Priority Date: 04/27/2011
• Status: Active Grant

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Mullick, Adam, Crooke, Rosanne M., Graham, Mark J., Dobie, Kenneth W., Lee, Richard
• Primary Examiner(s): Gibbs, Terra C
• Application Number:
  US14/114,187
• Publication Number:
  US 20140128453A1
• Time in Patent Office: 1,264 Days
• Field of Search: 514/44, 536/24.5
• US Class Current: 1/1
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• 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 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/06 Antiarrhythmics
• 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: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• 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 2310/341 : 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

Abstract

Provided herein are methods, compounds, and compositions for reducing expression of ApoCIII mRNA and protein in an animal. Also provided herein are methods, compounds, and compositions for increasing HDL levels and/or improving the ratio of TG to HDL and reducing plasma lipids and plasma glucose in an animal. Such methods, compounds, and compositions are useful to treat, prevent, delay, or ameliorate any one or more of cardiovascular disease or metabolic disorder, or a symptom thereof.

Claims

1. A method of increasing high density lipoprotein (HDL) levels in a human, comprising administering to a human in need thereof an antisense compound, wherein the antisense compound is a single-stranded oligonucleotide 12 to 30 nucleobases in length targeted to human apolipoprotein C-III (ApoCIII), whereby HDL levels are increased by at least 20% in the human.

2. The method of claim 1, wherein the oligonucleotide targeting ApoCIII is a modified single-stranded oligonucleotide.

3. The method of claim 2, wherein the modified oligonucleotide has a nucleobase sequence at least 90% or 100% complementary to a nucleobase sequence of SEQ ID NO:

1.

4. The method of claim 2, wherein the modified oligonucleotide consists of 20 linked nucleosides.

5. The method of claim 2, wherein at least one internucleoside linkage of the modified oligonucleotide is a modified internucleoside linkage, at least one nucleoside of the modified oligonucleotide comprises a modified sugar and/or at least one nucleoside of the modified oligonucleotide comprises a modified nucleobase.

6. The method of claim 5, wherein each modified internucleoside linkage of the modified oligonucleotide is a phosphorothioate internucleoside linkage.

7. The method of claim 5, wherein at least one modified sugar is a bicyclic sugar.

8. The method of claim 5, wherein at least one modified sugar comprises a 2′

-O-methyoxyethyl.

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

10. The method of claim 2, wherein the modified oligonucleotide comprises:

(a) a gap segment consisting of linked deoxynucleosides;

(b) a 5′

wing segment consisting of linked nucleosides;

(c) a 3′

wing segment consisting linked nucleosides;

wherein the gap segment is positioned immediately adjacent to and between the 5′

wing segment and the 3′

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

11. The method of claim 2, wherein the modified oligonucleotide comprises:

(a) a gap segment consisting of 10 linked deoxynucleosides;

(b) a 5′

wing segment consisting of 5 linked nucleosides;

(c) a 3′

wing segment consisting 5 linked nucleosides;

wherein the gap segment is positioned immediately adjacent to and between the 5′

wing segment and the 3′

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

-O-methyoxyethyl sugar, wherein each cytosine is a 5′

-methylcytosine, and wherein each internucleoside linkage is a phosphorothioate linkage.

12. The method of claim 1, wherein the compound is parenterally administered.

13. The method of claim 12, wherein the parenteral administration is subcutaneous administration.

14. The method of claim 1, further comprising administering a second agent.

15. The method of claim 14, wherein the second agent is selected from among siRNA targeting ApoCIII, an anti-ApoCIII antibody, an ApoCIII peptide inhibitor, an ApoCIII lowering agent, a cholesterol lowering agent, a non-HDL lipid lowering agent, a LDL lowering agent, a HDL raising agent, fish oil, a statin, niacin, a fibrate, DCCR (salt of diazoxide), an anti-diabetic agent, a glucose lowering agent and an anti-inflammatory agent.

16. A method of reducing chylomicronemia in a human, comprising administering to a human in need thereof an antisense compound, wherein the antisense compound is a single-stranded oligonucleotide 12 to 30 nucleobases in length targeted to human apolipoprotein C-III (ApoCIII), whereby chylomicronemia is reduced in the human.

17. The method of claim 16, wherein the antisense compound targeting ApoCIII is a modified single-stranded oligonucleotide.

18. The method of claim 17, wherein the modified oligonucleotide has a nucleobase sequence at least 90% or 100% complementary to a nucleobase sequence of SEQ ID NO:

1.

19. The method of claim 17, wherein the modified oligonucleotide consists of 20 linked nucleosides.

20. The method of claim 17, wherein at least one internucleoside linkage of the modified oligonucleotide is a modified internucleoside linkage, at least one nucleoside of the modified oligonucleotide comprises a modified sugar and/or at least one nucleoside of the modified oligonucleotide comprises a modified nucleobase.

21. The method of claim 20, wherein each modified internucleoside linkage of the modified oligonucleotide is a phosphorothioate internucleoside linkage.

22. The method of claim 20, wherein at least one modified sugar is a bicyclic sugar.

23. The method of claim 20, wherein at least one modified sugar comprises a 2′

-O-methyoxyethyl.

24. The method of claim 20, wherein the modified nucleobase is a 5-methylcytosine.

25. The method of claim 17, wherein the modified oligonucleotide comprises:

(g) a gap segment consisting of linked deoxynucleosides;

(h) a 5′

wing segment consisting of linked nucleosides;

(i) a 3′

wing segment consisting linked nucleosides;

wherein the gap segment is positioned immediately adjacent to and between the 5′

wing segment and the 3′

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

26. The method of claim 17, wherein the modified oligonucleotide comprises:

(g) a gap segment consisting of 10 linked deoxynucleosides;

(h) a 5′

wing segment consisting of 5 linked nucleosides;

(i) a 3′

wing segment consisting 5 linked nucleosides;

wherein the gap segment is positioned immediately adjacent to and between the 5′

wing segment and the 3′

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

-O-methyoxyethyl sugar, wherein each cytosine is a 5′

-methylcytosine, and wherein each internucleoside linkage is a phosphorothioate linkage.

27. The method of claim 16, wherein the compound is parenterally administered.

28. The method of claim 27, wherein the parenteral administration is subcutaneous administration.

29. The method of claim 16, further comprising administering a second agent.

30. The method of claim 29, wherein the second agent is selected from among siRNA targeting ApoCIII, an anti-ApoCIII antibody, an ApoCIII peptide inhibitor, an ApoCIII lowering agent, a cholesterol lowering agent, a non-HDL lipid lowering agent, a LDL lowering agent, a HDL raising agent, fish oil, a statin, niacin, a fibrate, DCCR (salt of diazoxide), an anti-diabetic agent, a glucose lowering agent and an anti-inflammatory agent.

31. A method of reducing pancreatitis in a human, comprising administering to a human in need thereof an antisense compound, wherein the antisense compound is a single-stranded oligonucleotide 12 to 30nucleobases in length targeted to human apolipoprotein C-III (ApoCIII), whereby pancreatitis is reduced in the human.

32. The method of claim 31, wherein the antisense compound targeting ApoCIII is a modified single-stranded oligonucleotide.

33. The method of claim 32, wherein the modified oligonucleotide has a nucleobase sequence at least 90% or 100% complementary to a nucleobase sequence of SEQ ID NO:

1.

34. The method of claim 32, wherein the modified oligonucleotide consists of 20 linked nucleosides.

35. The method of claim 32, wherein at least one internucleoside linkage of the modified oligonucleotide is a modified internucleoside linkage, at least one nucleoside of the modified oligonucleotide comprises a modified sugar and/or at least one nucleoside of the modified oligonucleotide comprises a modified nucleobase.

36. The method of claim 35, wherein each modified internucleoside linkage of the modified oligonucleotide is a phosphorothioate internucleoside linkage.

37. The method of claim 35, wherein at least one modified sugar is a bicyclic sugar.

38. The method of claim 35, wherein at least one modified sugar comprises a 2′

-O-methyoxyethyl.

39. The method of claim 35, wherein the modified nucleobase is a 5-methylcytosine.

40. The method of claim 32, wherein the modified oligonucleotide comprises:

(j) a gap segment consisting of linked deoxynucleosides;

(k) a 5′

wing segment consisting of linked nucleosides;

(l) a 3′

wing segment consisting linked nucleosides;

wherein the gap segment is positioned immediately adjacent to and 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 32, wherein the modified oligonucleotide comprises:

(j) a gap segment consisting of 10 linked deoxynucleosides;

(k) a 5′

wing segment consisting of 5 linked nucleosides;

(l) a 3′

wing segment consisting 5 linked nucleosides;

wherein the gap segment is positioned immediately adjacent to and between the 5′

wing segment and the 3′

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

-O-methyoxyethyl sugar, wherein each cytosine is a 5′

-methylcytosine, and wherein each internucleoside linkage is a phosphorothioate linkage.

42. The method of claim 31, wherein the compound is parenterally administered.

43. The method of claim 42, wherein the parenteral administration is subcutaneous administration.

44. The method of claim 31, further comprising administering a second agent.

45. The method of claim 44, wherein the second agent is selected from among siRNA targeting ApoCIII, an anti-ApoCIII antibody, an ApoCIII peptide inhibitor, an ApoCIII lowering agent, a cholesterol lowering agent, a non-HDL lipid lowering agent, a LDL lowering agent, a HDL raising agent, fish oil, a statin, niacin, a fibrate, DCCR (salt of diazoxide), an anti-diabetic agent, a glucose lowering agent and an anti-inflammatory agent.