Modulation of apolipoprotein C-III expression

• US 20040208856A1
• Filed: 04/16/2003
• Published: 10/21/2004
• Est. Priority Date: 04/16/2003
• Status: Active Grant

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Inventors: Dobie, Kenneth W., Graham, Mark J., Crooke, Rosanne M., Lemonidis, Kristina M.
• Granted Patent: US 7,598,227 B2
• Time in Patent Office: Days
• US Class Current: 424/93.21
• 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 compound 8 to 80 nucleobases in length targeted to a nucleic acid molecule encoding apolipoprotein C-III, wherein said compound specifically hybridizes with said nucleic acid molecule encoding apolipoprotein C-III (SEQ ID NO:

4) and inhibits the expression of apolipoprotein C-III.

2. The compound of claim 1 comprising 12 to 50 nucleobases in length.

3. The compound of claim 2 comprising 15 to 30 nucleobases in length.

4. The compound of claim 1 comprising an oligonucleotide.

5. The compound of claim 4 comprising an antisense oligonucleotide.

6. The compound of claim 4 comprising a DNA oligonucleotide.

7. The compound of claim 4 comprising an RNA oligonucleotide.

8. The compound of claim 4 comprising a chimeric oligonucleotide.

9. The compound of claim 4 wherein at least a portion of said compound hybridizes with RNA to form an oligonucleotide-RNA duplex.

10. The compound of claim 1 having at least 70% complementarity with a nucleic acid molecule encoding apolipoprotein C-III (SEQ ID NO:

4) said compound specifically hybridizing to and inhibiting the expression of apolipoprotein C-III.

11. The compound of claim 1 having at least 80% complementarity with a nucleic acid molecule encoding apolipoprotein C-III (SEQ ID NO:

4) said compound specifically hybridizing to and inhibiting the expression of apolipoprotein C-III.

12. The compound of claim 1 having at least 90% complementarity with a nucleic acid molecule encoding apolipoprotein C-III (SEQ ID NO:

4) said compound specifically hybridizing to and inhibiting the expression of apolipoprotein C-III.

13. The compound of claim 1 having at least 95% complementarity with a nucleic acid molecule encoding apolipoprotein C-III (SEQ ID NO:

4) said compound specifically hybridizing to and inhibiting the expression of apolipoprotein C-III.

14. The compound of claim 1 having at least one modified internucleoside linkage, sugar moiety, or nucleobase.

15. The compound of claim 1 having at least one 2′

-O-methoxyethyl sugar moiety.

16. The compound of claim 1 having at least one phosphorothioate internucleoside linkage.

17. The compound of claim 1 having at least one 5-methylcytosine.

18. A method of inhibiting the expression of apolipoprotein C-III in cells or tissues comprising contacting said cells or tissues with the compound of claim 1 so that expression of apolipoprotein C-III is inhibited.

22. A kit or assay device comprising the compound of claim 1.

23. A method of treating an animal having a disease or condition associated with apolipoprotein C-III comprising administering to said animal a therapeutically or prophylactically effective amount of the compound of claim 1 so that expression of apolipoprotein C-III is inhibited.

24. The method of claim 23 wherein the condition involves abnormal lipid metabolism.

25. The method of claim 23 wherein the condition involves abnormal cholesterol metabolism.

26. The method of claim 23 wherein the condition is atherosclerosis.

27. The method of claim 23 wherein the condition is an abnormal metabolic condition.

28. The method of claim 27 wherein the abnormal metabolic condition is hyperlipidemia.

29. The method of claim 23 wherein the disease is diabetes.

30. The method of claim 29 wherein the diabetes is Type 2 diabetes.

31. The method of claim 23 wherein the condition is obesity.

32. The method of claim 23 wherein the disease is cardiovascular disease.

33. A method of modulating glucose levels in an animal comprising administering to said animal the compound of claim 1.

34. The method of claim 33 wherein the animal is a human.

35. The method of claim 33 wherein the glucose levels are plasma glucose levels.

36. The method of claim 33 wherein the glucose levels are serum glucose levels.

37. The method of claim 33 wherein the animal is a diabetic animal.

38. A method of preventing 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 the compound of claim 1.

39. The method of claim 38 wherein the animal is a human.

40. The method of claim 38 wherein the condition is an abnormal metabolic condition.

41. The method of claim 40 wherein the abnormal metabolic condition is hyperlipidemia.

42. The method of claim 38 wherein the disease is diabetes.

43. The method of claim 42 wherein the diabetes is Type 2 diabetes.

44. The method of claim 38 wherein the condition is obesity.

45. A method of lowering cholesterol levels in an animal comprising administering to said animal the compound of claim 1.

46. The method of claim 45 wherein the animal is a human.

47. The method of claim 45 wherein the cholesterol levels are plasma cholesterol levels.

48. The method of claim 45 wherein the cholesterol levels are serum cholesterol levels.

49. A method of lowering triglyceride levels in an animal comprising administering to said animal the compound of claim 1.

50. The method of claim 49 wherein the animal is a human.

51. The method of claim 49 wherein the triglyceride levels are plasma triglyceride levels.

52. The method of claim 49 wherein the triglyceride levels are serum triglyceride levels.

19. A method of screening for a modulator of apolipoprotein C-III, the method comprising the steps of:

a. contacting a preferred target segment of a nucleic acid molecule encoding apolipoprotein C-III with one or more candidate modulators of apolipoprotein C-III, and

b. identifying one or more modulators of apolipoprotein C-III expression which modulate the expression of apolipoprotein C-III.

20. The method of claim 19 wherein the modulator of apolipoprotein C-III expression comprises an oligonucleotide, an antisense oligonucleotide, a DNA oligonucleotide, an RNA oligonucleotide, an RNA oligonucleotide having at least a portion of said RNA oligonucleotide capable of hybridizing with RNA to form an oligonucleotide-RNA duplex, or a chimeric oligonucleotide.

21. A diagnostic method for identifying a disease state comprising identifying the presence of apolipoprotein C-III in a sample using at least one of the primers comprising SEQ ID NOs 5 or 6, or the probe comprising SEQ ID NO 7.