Modulation of apolipoprotein C-III expression

• US 7,598,227 B2
• Filed: 04/16/2003
• Issued: 10/06/2009
• Est. Priority Date: 04/16/2003
• Status: Expired due to Term

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Graham, Mark J., Lemonidis, Kristina M., Dobie, Kenneth W., Crooke, Rosanne M.
• Primary Examiner(s): McGarry; Sean
• Assistant Examiner(s): GIBBS, TERRA C
• Application Number:
  US10/418,780
• Publication Number:
  US 20040208856A1
• Time in Patent Office: 2,365 Days
• Field of Search: 514/44, 536/24.5
• US Class Current: 514/44.00R
• 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 treating an animal having hyperlipidemia, said method comprising:

administering to an animal suffering from hyperlipidemia an antisense compound comprising an antisense oligonucleotide, wherein said antisense oligonucleotide specifically hybridizes within nucleotides 3253-3558 of SEQ ID NO;

4 and consists of 15 to 30 linked nucleosides, thereby reducing hyperlipidemia in said animal.

12. The method of any one of claims 1, 2, 4 or 8, wherein said antisense oligonucleotide comprises at least one modified internucleoside linkage, sugar moiety, or nucleobase.

13. The method of claim 12, wherein said modified internucleoside linkage is a phosphorothioate linkage.

14. The method of claim 12, wherein said modified sugar moiety is a 2′

-O-methoxyethyl sugar moiety.

15. The method of claim 12, wherein said modified nucleobase is a 5-methylcytosine.

16. The method of any one of claims 1, 2, 4 or 8, wherein said antisense compound consists of an antisense oligonucleotide.

17. The method of any one of claims 1, 2, 4, or 8, wherein said antisense oligonucleotide is fully complementary to SEQ ID NO:

4.

18. The method of any one of claims 1, 2, 4, or 8, wherein said antisense oligonucleotide consists of 18, 19, 20, 21, or 22 linked nucleosides.

19. The method of any one of claims 1, 2, 4, or 8, wherein said antisense oligonucleotide consists of 20 linked nucleosides.

20. The method of any one of claims 1, 2, 4, or 8, wherein said antisense compound demonstrates a reduction in apolipoprotein C-III mRNA levels of at least 45% when applied in vitro to cultured HepG2 cells at a concentration of 50 to 300 nM.

21. The method of any one of claims 1, 2, 4, or 8, wherein said antisense oligonucleotide comprises a plurality of 2′

-deoxynucleotides flanked on each side by at least one 2′

-O-methoxyethyl nucleotide.

22. The method of claim 1, wherein said animal is a human.

23. The method of claim 1, 2, 4, or 8, wherein said antisense oligonucleotide specifically hybridizes within nucleotides 3253-3380, 3417-3445, 3445-3491, or 3509-3558 of SEQ ID NO:

4.

24. The method of claim 1, 2, 4, or 8, wherein said antisense oligonucleotide consists of:

a gap segment consisting often linked 2′

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

25. The method of claim 24, wherein said antisense oligonucleotide comprises at least one cytosine that is a 5-methylcytosine.

26. The method of claim 25, wherein each cytosine is a 5-methylcytosine.

27. The method of claim 26, wherein the antisense oligonucleotide is fully complementary to SEQ ID NO:

4.

28. The method of claim 1, 2, 4, or 8, further comprising measuring serum apolipoprotein C-III levels in the serum of said animal.

2. A method of delaying the onset of hyperlipidemia in an animal, said method comprising:

administering to an animal at risk for developing hyperlipidemia an antisense compound comprising an antisense oligonucleotide, wherein said antisense oligonucleotide specifically hybridizes within nucleotides 3253-3558 of SEQ ID NO;

4 and consists of 15 to 30 linked nucleosides, thereby delaying the onset of hyperlipidemia.

3. The method of claim 2 wherein the animal is a human.

4. A method of lowering cholesterol levels in an animal comprising:

administering to an animal in need of a reduction in cholesterol levels an antisense compound comprising an antisense oligonucleotide, wherein said antisense oligonucleotide specifically hybridizes within nucleotides 3253-3558 of SEQ ID NO;

4 and consists of 15 to 30 linked nucleosides, wherein said administering results in reduced cholesterol levels in said animal.

5. The method of claim 4 wherein the animal is a human.

6. The method of claim 4 wherein the cholesterol levels are plasma cholesterol levels.

7. The method of claim 4 wherein the cholesterol levels are serum cholesterol levels.

8. A method of lowering triglyceride levels in an animal comprising:

administering to an animal in need of a reduction in triglyceride levels an antisense compound comprising an antisense oligonucleotide, wherein said antisense oligonucleotide specifically hybridizes within nucleotides 3253-3558 of SEQ ID NO;

4 and consists of 15 to 30 linked nucleosides, wherein said administering results in reduced triglyceride levels in said animal.

9. The method of claim 8 wherein the animal is a human.

10. The method of claim 8 wherein the triglyceride levels are plasma triglyceride levels.

11. The method of claim 8 wherein the triglyceride levels are serum triglyceride levels.