Enhanced antisense oligonucleotides

• US 7,919,472 B2
• Filed: 09/19/2005
• Issued: 04/05/2011
• Est. Priority Date: 09/17/2004
• Status: Expired due to Fees

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Monia, Brett P., Bhanot, Sanjay, Siwkowski, Andrew M.
• Primary Examiner(s): ZARA, JANE J
• Application Number:
  US11/231,243
• Publication Number:
  US 20060063730A1
• Time in Patent Office: 2,024 Days
• Field of Search: 435/6, 435/91.1, 435/91.31, 435/455, 514/44, 536/23.1, 536/24.5, 536/24.31
• US Class Current: N/A
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• 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 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• 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: A61P 9/12 Antihypertensives
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• 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 2310/3525 MOE, methoxyethoxy
• CPC Class Codes: C12N 2320/50 Methods for regulating/modulating their activity

Abstract

Described herein are gap-widened antisense oligonucleotides having improved therapeutic index as compared to 5-10-5 MOE gapmer antisense oligonucleotides of the same sequence. Also described are methods of reducing a target RNA in an animal using the gap-widened antisense oligonucleotides of the present invention. Further, are methods for selecting a gap-widened antisense oligonucleotides.

Claims

1. A method of reducing expression of a target RNA in an animal, in need of reducing expression of said target RNA, comprising administering to said animal a gap-widened antisense oligonucleotide 18-24 nucleotides in length targeting a preselected RNA target in the liver, wherein said oligonucleotide comprises:

(a) a gap region having 12 to 18 contiguous 2′

-deoxyribonucleotides;

(b) a first wing region having 2 to 4 contiguous nucleosides; and

(c) a second wing region having 2 to 4 contiguous nucleosides;

wherein the gap region is located between said first wing region and said second wing region and, wherein each nucleoside of said first and second wing regions comprises a 2′

-O-(2-methoxyethyl) modification;

thereby reducing expression of said preselected RNA target in the liver of said animal.

2. The method of claim 1, wherein the target RNA is associated with a metabolic or a cardiovascular disease or condition.

3. The method of claim 1, wherein the metabolic disease or condition is selected from metabolic syndrome, diabetes, obesity, hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, Type 2 diabetes, diet-induced obesity, hyperglycemia, insulin resistance, hepatic steatosis, fatty liver disease, or non-alcoholic steatohepatitis.

4. The method of claim 1, wherein the cardiovascular disease or condition is selected from familial hypercholesterolemia, nonfamilial hypercholesterolemia, mixed dyslipidemia, dysbetalipoproteinemia, atherosclerosis, coronary artery disease, myocardial infarction, hypertension, carotid artery disease, stroke, cerebrovascular disease, peripheral vascular disease, thrombosis, or arterial aneurism.

5. The method of claim 1, wherein the gap-widened antisense oligonucleotide has at least one phosphorothioate internucleotide linkage.

6. The method of claim 5, wherein the gap-widened antisense oligonucleotide has all phosphorothioate internucleotide linkages.

7. The method of claim 1, wherein gap-widened antisense oligonucleotide has at least one 5-methylcytosine.

9. The method of claim 1, wherein said wing-gap-wing motif is selected from 2-16-2, 3-14-3, 2-14-2, 3-12-3 and 4-12-4.

10. The method of claim 9, wherein said wing-gap-wing motif is 2-16-2.

11. The method of claim 9, wherein said wing-gap-wing motif is 3-14-3.

12. The method of claim 9, wherein said wing-gap-wing motif is 2-14-2.

13. The method of claim 9, wherein said wing-gap-wing motif is 3-12-3.

14. The method of claim 9, wherein said wing-gap-wing motif is 4-12-4.

8. A method of modulating gene expression in an animal comprising the step of contacting said animal with a pharmaceutical composition comprising a gap-widened antisense oligonucleotide 18-24 nucleotides in length targeting a preselected RNA target in the liver in the manufacture of a medicament for the treatment of disorders and diseases related to said RNA, wherein the antisense oligonucleotide comprises:

(a) a gap region having 12 to 18 contiguous 2′

-deoxyribonucleotides;

(b) a first wing region having 2 to 4 contiguous nucleosides; and

(c) a second wing region having 2 to 4 contiguous nucleosides;

wherein the gap region is located between said first wing region and said second wing region and, wherein each nucleoside of said first and second wing regions comprises a 2′

-O-(2-methoxyethyl) modification;

thereby modulating the expression of said preselected RNA target in the liver of said animal.

15. The method of claim 8, wherein said wing-gap-wing motif is selected from 2-16-2, 3-14-3, 2-14-2, 3-12-3 and 4-12-4.

16. The method of claim 15, wherein said wing-gap-wing motif is 2-16-2.

17. The method of claim 15, wherein said wing-gap-wing motif is 3-14-3.

18. The method of claim 15, wherein said wing-gap-wing motif is 2-14-2.

19. The method of claim 15, wherein said wing-gap-wing motif is 3-12-3.

20. The method of claim 15, wherein said wing-gap-wing motif is 4-12-4.