Modulation of apolipoprotein C-III expression

• US 9,624,496 B2
• Filed: 04/29/2016
• Issued: 04/18/2017
• Est. Priority Date: 04/16/2003
• Status: Expired due to Term

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: Ionis Pharmaceuticals, Inc.
• Inventors: Crooke, Rosanne M., Graham, Mark J., Dobie, Kenneth W.
• Primary Examiner(s): Gibbs, Terra C
• Application Number:
  US15/143,222
• Publication Number:
  US 20160237434A1
• Time in Patent Office: 354 Days
• Field of Search: 536 231, 536 243, 536 245
• 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 a modified antisense oligonucleotide, wherein said modified 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 (2′

-MOE), 2′

-fluoro (2′

-F) or 2′

-methoxy (2′

-O—

CH₃) 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 a modified antisense oligonucleotide.

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

4.

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

9. The compound of claim 1, wherein said modified 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 modified 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 modified antisense oligonucleotide specifically hybridizes within nucleotides 3253-3380, 3417-3445, 3445-3491, 3451-3500 or 3509-3558 of SEQ ID NO:

4.

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

4.

17. The compound of claim 1, wherein said modified antisense oligonucleotide comprises at least 8 consecutive nucleobases selected from the nucleobase sequence of SEQ ID NO:

78.

18. The compound of claim 1, wherein said modified antisense oligonucleotide is in salt form.

19. A composition comprising the compound of claim 1 and a pharmaceutically acceptable carrier or diluent.

20. The compound of claim 1, wherein said compound is single-stranded or double-stranded.

21. The compound of claim 1, further comprising a conjugate group.

22. The compound of claim 1, wherein said modified antisense oligonucleotide consists of the nucleobase sequence of SEQ ID NO:

87.