Modulation of apolipoprotein C-III expression

• US 8,530,439 B2
• Filed: 05/17/2010
• Issued: 09/10/2013
• Est. Priority Date: 04/16/2003
• Status: Expired due to Term

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Crooke, Rosanne M., Graham, Mark J., Lemonidis, Kristina M., Dobie, Kenneth W.
• Primary Examiner(s): GIBBS, TERRA C
• Application Number:
  US12/781,479
• Publication Number:
  US 20110060030A1
• Time in Patent Office: 1,212 Days
• Field of Search: 514/44, 536/24.5
• US Class Current: 514/44.A
• 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 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 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: 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 2310/314 Phosphoramidates
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• 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 2310/3521 Methyl
• CPC Class Codes: C12N 2320/30 Special therapeutic applications
• CPC Class Codes: C12N 2330/31 Libraries, arrays

Abstract

Compounds, compositions and methods are provided for modulating the expression of apolipoprotein C-III. The compositions comprise oligonucleotides, targeted to nucleic acid encoding apolipoprotein C-III. Methods of using these compounds for modulation of apolipoprotein C-III expression and for diagnosis and treatment of disease associated with expression of apolipoprotein C-III are provided.

Claims

1. A method of inhibiting the expression of apolipoprotein C-III in cells or tissues comprising contacting said cells or tissues with a compound 10 to 25 nucleobases in length targeted to a nucleic acid molecule encoding apolipoprotein C-III, wherein said compound specifically hybridizes with a 5′

untranslated region or coding region of said nucleic acid molecule encoding apolipoprotein C-III (SEQ ID NO;

     4), so that expression of apolipoprotein C-III is inhibited by at least 45%.

7. The method of claim 1, wherein the compound is 20 nucleobases in length.

8. The method of claim 1, wherein the compound is 20 nucleotides in length, comprising ten 2′

-deoxynucleotides, flanked on each side by five 2′

-methoxyethyl nucleotides, wherein all internucleoside linkages are phosphorothioate, and all cytosine residues are 5-methylcytosines.

9. The method of claim 1, wherein the compound comprises at least one modified internucleoside linkage, sugar moiety, or nucleobase.

10. The method of claim 1, wherein the compound comprises at least one 2′

-O-methoxyethyl sugar moiety.

11. The method of claim 1, wherein the compound comprises at least one phosphorothioate internucleoside linkage.

12. The method of claim 1, wherein the compound comprises at least one 5-methylcytosine.

2. A method of treating an animal having a disease or condition associated with apolipoprotein C-III or delaying the onset of a disease or condition associated with apolipoprotein C-III in an animal comprising administering to said animal a therapeutically or prophylactically effective amount of a compound 10 to 25 nucleobases in length targeted to a nucleic acid molecule encoding apolipoprotein C-III, wherein said compound specifically hybridizes with a 5′

untranslated region or coding region of said nucleic acid molecule encoding apolipoprotein C-III (SEQ ID NO;

     4), so that expression of apolipoprotein C-III is inhibited by at least 45%, thereby treating an animal having a disease or condition associated with apolipoprotein C-III or delaying the onset of a disease or condition associated with apolipoprotein C-III in an animal.

3. The method of claim 2, wherein the condition involves abnormal lipid metabolism, abnormal cholesterol metabolism, atherosclerosis, abnormal metabolic condition, hyperlipidemia, diabetes, Type 2 diabetes, obesity, hypertriglyceridemia, and/or cardiovascular disease.

13. The method of claim 2, wherein the compound is 20 nucleobases in length.

14. The method of claim 2, wherein the compound is 20 nucleotides in length, comprising ten 2′

-deoxynucleotides, flanked on each side by five 2′

-methoxyethyl nucleotides, wherein all internucleoside linkages are phosphorothioate, and all cytosine residues are 5-methylcytosines.

15. The method of claim 2, wherein the compound comprises at least one modified internucleoside linkage, sugar moiety, or nucleobase.

16. The method of claim 2, wherein the compound comprises at least one 2′

-O-methoxyethyl sugar moiety.

17. The method of claim 2, wherein the compound comprises at least one phosphorothioate internucleoside linkage.

18. The method of claim 2, wherein the compound comprises at least one 5-methylcytosine.

4. A method of modulating glucose levels in an animal comprising administering to said animal a compound 10 to 25 nucleobases in length targeted to a nucleic acid molecule encoding apolipoprotein C-III, wherein said compound specifically hybridizes with a 5′

untranslated region or coding region of said nucleic acid molecule encoding apolipoprotein C-III (SEQ ID NO;

     4), so that expression of apolipoprotein C-III is inhibited by at least 45%, thereby modulating glucose levels in an animal.

5. The method of claim 4, wherein serum glucose levels are reduced.

6. The method of claim 4, wherein fasted serum insulin levels in an animal are decreased.

19. The method of claim 4, wherein the compound is 20 nucleobases in length.

20. The method of claim 4, wherein the compound is 20 nucleotides in length, comprising ten 2′

-deoxynucleotides, flanked on each side by five 2′

-methoxyethyl nucleotides, wherein all internucleoside linkages are phosphorothioate, and all cytosine residues are 5-methylcytosines.

21. The method of claim 4, wherein the compound comprises at least one modified internucleoside linkage, sugar moiety, or nucleobase.

22. The method of claim 4, wherein the compound comprises at least one 2′

-O-methoxyethyl sugar moiety.

23. The method of claim 4, wherein the compound comprises at least one phosphorothioate internucleoside linkage.

24. The method of claim 4, wherein the compound comprises at least one 5-methylcytosine.

25. A method of inhibiting the expression of apolipoprotein C-III in cells or tissues comprising contacting said cells or tissues with a compound 10 to 25 nucleobases in length targeted to a nucleic acid molecule encoding apolipoprotein C-III, wherein said compound comprises at least an 8 consecutive nucleobase portion of SEQ ID NO:

19, 20, 21, 22, 23, 25, 27, 28, 29, 30, 31, 32, 33, 34, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 55, 56, 57, 58, 59, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95 or 96, and wherein said compound specifically hybridizes with said nucleic acid molecule encoding apolipoprotein C-III, so that expression of apolipoprotein C-III is inhibited.

31. The method of claim 25, wherein the compound is 20 nucleobases in length.

32. The method of claim 25, wherein the compound is 20 nucleotides in length, comprising ten 2′

-deoxynucleotides, flanked on each side by five 2′

-methoxyethyl nucleotides, wherein all internucleoside linkages are phosphorothioate, and all cytosine residues are 5-methylcytosines.

33. The method of claim 25, wherein the compound comprises at least one modified internucleoside linkage, sugar moiety, or nucleobase.

34. The method of claim 25, wherein the compound comprises at least one 2′

-O-methoxyethyl sugar moiety.

35. The method of claim 25, wherein the compound comprises at least one phosphorothioate internucleoside linkage.

36. The method of claim 25, wherein the compound comprises at least one 5-methylcytosine.

26. A method of treating an animal having a disease or condition associated with apolipoprotein C-III or delaying the onset of a disease or condition associated with apolipoprotein C-III in an animal comprising administering to said animal a therapeutically or prophylactically effective amount of a compound 10 to 25 nucleobases in length targeted to a nucleic acid molecule encoding apolipoprotein C-III, wherein said compound comprises at least an 8 consecutive nucleobase portion of SEQ ID NO:

19, 20, 21, 22, 23, 25, 27, 28, 29, 30, 31, 32, 33, 34, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 55, 56, 57, 58, 59, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95 or 96, and wherein said compound specifically hybridizes with said nucleic acid molecule encoding apolipoprotein C-III, so that expression of apolipoprotein C-III is inhibited, thereby treating an animal having a disease or condition associated with apolipoprotein C-III or delaying the onset of a disease or condition associated with apolipoprotein C-III in an animal.

27. The method of claim 26, wherein the condition involves abnormal lipid metabolism, abnormal cholesterol metabolism, atherosclerosis, abnormal metabolic condition, hyperlipidemia, diabetes, Type 2 diabetes, obesity, hypertriglyceridemia, and/or cardiovascular disease.

37. The method of claim 26, wherein the compound is 20 nucleobases in length.

38. The method of claim 26, wherein the compound is 20 nucleotides in length, comprising ten 2′

-deoxynucleotides, flanked on each side by five 2′

-methoxyethyl nucleotides, wherein all internucleoside linkages are phosphorothioate, and all cytosine residues are 5-methylcytosines.

39. The method of claim 26, wherein the compound comprises at least one modified internucleoside linkage, sugar moiety, or nucleobase.

40. The method of claim 26, wherein the compound comprises at least one 2′

-O-methoxyethyl sugar moiety.

41. The method of claim 26, wherein the compound comprises at least one phosphorothioate internucleoside linkage.

42. The method of claim 26, wherein the compound comprises at least one 5-methylcytosine.

28. A method of modulating glucose levels in an animal comprising administering to said animal a compound 10 to 25 nucleobases in length targeted to a nucleic acid molecule encoding apolipoprotein C-III, wherein said compound comprises at least an 8 consecutive nucleobase portion of SEQ ID NO:

19, 20, 21, 22, 23, 25, 27, 28, 29, 30, 31, 32, 33, 34, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 55, 56, 57, 58, 59, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95 or 96, and wherein said compound specifically hybridizes with said nucleic acid molecule encoding apolipoprotein C-III, so that expression of apolipoprotein C-III is inhibited, thereby modulating glucose levels in an animal.

29. The method of claim 28, wherein serum glucose levels are reduced.

30. The method of claim 28, wherein fasted serum insulin levels in an animal are decreased.

43. The method of claim 28, wherein the compound is 20 nucleobases in length.

44. The method of claim 28, wherein the compound is 20 nucleotides in length, comprising ten 2′

-deoxynucleotides, flanked on each side by five 2′

-methoxyethyl nucleotides, wherein all internucleoside linkages are phosphorothioate, and all cytosine residues are 5-methylcytosines.

45. The method of claim 28, wherein the compound comprises at least one modified internucleoside linkage, sugar moiety, or nucleobase.

46. The method of claim 28, wherein the compound comprises at least one 2′

-O-methoxyethyl sugar moiety.

47. The method of claim 28, wherein the compound comprises at least one phosphorothioate internucleoside linkage.

48. The method of claim 28, wherein the compound comprises at least one 5-methylcytosine.