Modulation of apolipoprotein C-III expression

• US 9,365,848 B2
• Filed: 08/07/2013
• Issued: 06/14/2016
• Est. Priority Date: 04/16/2003
• Status: Expired due to Fees

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: Ionis Pharmaceuticals, Inc.
• Inventors: Crooke, Rosanne M., Graham, Mark J., Lemonidis, Kristina M.
• Primary Examiner(s): Gibbs, Terra C
• Application Number:
  US13/961,646
• Publication Number:
  US 20130317085A1
• Time in Patent Office: 1,042 Days
• Field of Search: 435/6.1, 435/91.1, 435/325, 536/24.5
• US Class Current: 1/1
• 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 for identifying a modified antisense compound having the potential for treating a subject with a disease or condition associated with the expression of apolipoprotein C-III levels, comprising:

(a) obtaining a modified antisense compound comprising at least one modified internucleoside linkage, sugar moiety or nucleobase, wherein the modified antisense compound targets a nucleic acid region corresponding to the 5′

cap site, the 5′

untranslated region, the translation initiation codon region, the coding region, the translation termination codon region or the 3′

untranslated region of apolipoprotein C-III;

(b) treating an animal with the antisense compound; and

(c) comparing the amount of reduction in apolipoprotein C-III expression levels in the treated animal to apolipoprotein C-III expression levels in an untreated animal;

wherein, if the reduction of levels in the treated animal in (c) is at least 50%, the antisense compound is identified as having the potential for treating a subject with a disease or condition associated with the expression of apolipoprotein C-III.

2. The method of claim 1, wherein the modified antisense compound additionally is tested for its ability to reduce apolipoprotein C-III levels by a method comprising:

(1) treating a cell, tissue or organ with the modified antisense compound; and

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

thereby determining whether the antisense compound has the ability to reduce apolipoprotein C-III levels in the cell, tissue or organ.

3. The method of claim 2, wherein a cell is treated in (1), and the cell is HepG2.

4. The method of claim 2, wherein a cell is treated in (1), and the cell is a monkey cell.

5. The method of claim 2, wherein a cell is treated in (1), and the cell is a hepatocyte.

6. The method of claim 2, wherein a tissue is treated in (1), and the tissue is blood.

7. The method of claim 2, wherein there is at least a 45% reduction in apolipoprotein C-III mRNA levels after (1) and (2).

8. The method of claim 2, wherein the antisense compound is administered in (1) at a concentration ranging from 50 to 300 nM.

9. The method of claim 1, wherein the animal is a mouse.

10. The method of claim 1, wherein the animal is a rat.

11. The method of claim 1, wherein the subject is a human.

12. The method of claim 1, further comprising determining whether the antisense compound reduces triglyceride, glucose and/or cholesterol levels in an animal when the antisense compound is administered to the animal, compared to the levels in the absence of the antisense compound.

13. The method of claim 1, wherein the reduction in apolipoprotein C-III expression levels comprises a reduction in mRNA levels of apolipoprotein C-III.

14. The method of claim 1, wherein the modified antisense compound targets a nucleic acid region corresponding to the coding region or the 3′

untranslated region of apolipoprotein C-III.

15. The method of claim 1, further comprising determining whether the antisense compound is toxic to the animal.

16. The method of claim 1, further comprising determining whether the antisense compound increases ALT and/or AST liver enzyme levels.