MODULATION OF APOLIPOPROTEIN C-III EXPRESSION

• US 20130317085A1
• Filed: 08/07/2013
• Published: 11/28/2013
• Est. Priority Date: 04/16/2003
• Status: Active Grant

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Crooke, Rosanne M., Graham, Mark J., Lemonidis, Kristina M., Dobie, Kenneth W.
• Granted Patent: US 9,365,848 B2
• Time in Patent Office: Days
• 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 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.

2. The compound of claim 1, wherein said compound comprises at least one modified internucleoside linkage, sugar moiety, or nucleobase.

3. The compound of claim 2, wherein said modified internucleoside linkage is a phosphorothioate linkage.

4. The compound of claim 2, wherein said modified sugar moiety is a 2′

-O-methoxyethyl sugar moiety.

5. The compound of claim 2, wherein said modified nucleobase is a 5-methylcytosine.

6. The compound of claim 1, wherein said compound consists of an antisense oligonucleotide.

7. The compound of claim 1, wherein said antisense oligonucleotide is fully complementary to SEQ ID NO:

4.

8. The compound of claim 1, wherein said antisense oligonucleotide consists of 18, 19, 20, 21, or 22 linked nucleosides.

9. The compound of claim 1, wherein said antisense oligonucleotide consists of 20 linked nucleosides.

10. The compound of claim 1, wherein said 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.

11. The compound of claim 1, wherein said antisense oligonucleotide comprises a plurality of 2′

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

-O-methoxyethyl nucleotide.

12. The compound of claim 1, wherein said antisense oligonucleotide specifically hybridizes within nucleotides 3253-3380, 3417-3445, 3445-3491, or 3509-3558 of SEQ ID NO:

4.

13. The compound of claim 1, 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.

14. The compound of claim 13, wherein said antisense oligonucleotide comprises at least one cytosine that is a 5-methylcytosine.

15. The compound of claim 14, wherein each cytosine is a 5-methylcytosine.

16. The compound of claim 15, wherein the antisense oligonucleotide is fully complementary to SEQ ID NO:

4.

17. A method for identifying an antisense compound for reducing apolipoprotein C-III levels comprising

(a) treating a cell, tissue, organ or animal with the antisense compound;

(b) comparing the amount of reduction in apolipoprotein C-III levels in the treated cell, tissue, organ or animal to apolipoprotein C-III levels in an untreated cell, tissue, organ or animal;

thereby, identifying the antisense compound that reduces apolipoprotein C-III levels.

18. The method of claim 17, wherein the identification is performed in vitro, in vivo or ex vivo.

19. The method of claim 17, wherein the cell is HepG2 cells.

20. The method of claim 17, wherein the tissue is blood.

21. The method of claim 17, wherein the reduction in apolipoprotein C-III mRNA levels is at least 45%.

22. The method of claim 17, wherein the antisense compound is administered at a concentration ranging from 50 to 300 nM.

23. The method of claim 17, wherein the antisense compound comprises at least one modified internucleoside linkage, sugar moiety, or nucleobase.

24. A method for reducing apolipoprotein C-III levels in a cell, tissue, organ or animal comprising administering an antisense compound to the cell, tissue, organ or animal, wherein the antisense compound specifically hybridizes to apolipoprotein C-III and wherein the antisense compound reduces apolipoprotein C-III levels.

25. The method of claim 24, wherein the reduction is performed in vitro, in vivo or ex vivo.

26. The method of claim 24, wherein the cell is HepG2 cells.

27. The method of claim 24, wherein the tissue is blood.

28. The method of claim 24, wherein the reduction in apolipoprotein C-III mRNA levels is at least 45%.

29. The method of claim 24, wherein the antisense compound is administered at a concentration ranging from 50 to 300 nM.

30. The method of claim 24, wherein the antisense compound comprises at least one modified internucleoside linkage, sugar moiety, or nucleobase.