RNAi Inhibition of Alpha-ENaC Expression

• US 20090156529A1
• Filed: 06/16/2008
• Published: 06/18/2009
• Est. Priority Date: 06/15/2007
• Status: Active Grant

• Current Assignee: Arrowhead Pharmaceuticals, Inc.
• Original Assignee: Novartis Ag
• Inventors: TAN, Pamela, VAN HEEKE, Gino, GEICK, Anke, VORNLOCHER, Hans-Peter, HICKMAN, Emma, DANAHAY, Henry Luke
• Granted Patent: US 7,718,632 B2
• Time in Patent Office: Days
• US Class Current: 514/44.R
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 3/12 for electrolyte homeostasis
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/12 Antihypertensives
• CPC Class Codes: C12N 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/14 interfering N.A.
• 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 2310/351 Conjugate
• CPC Class Codes: C12N 2310/3513 Protein; Peptide
• CPC Class Codes: C12N 2310/3515 : Lipophilic moiety, e.g. cholesterol
• CPC Class Codes: C12N 2310/3521 : Methyl

Abstract

The invention relates to compositions and methods for modulating the expression of alpha-ENaC, and more particularly to the downregulation of alpha-ENaC expression by chemically modified oligonucleotides.

Claims

1. An iRNA agent comprising a sense strand, wherein the sense strand comprises at least 15 contiguous nucleotides that differ by no more than 1, 2, or 3 nucleotides from the sense strand sequences of any one of the agents provided in Tables 1A-1D, agents numbered ND8285-ND10788, and an antisense strand, wherein the antisense strand comprises at least 15 contiguous nucleotides that differ by no more than 1, 2, or 3 nucleotides from the antisense sequences of any one of the agents provided in Tables 1A-1D, agents numbered ND8285-ND10788.

4. An iRNA agent according to claim 1 where the agent is selected from ND-8302, ND-8332, ND-8348, ND-8356, ND-8357, ND-8373, ND-8381, ND-8396, ND-8450 and ND-8453.

5. An iRNA agent according to claim 1 where the agent is selected from ND-8356, ND-8357 and ND-8396.

6. The iRNA agent of claim 1, wherein the antisense RNA strand is 30 or fewer nucleotides in length, and the duplex region of the iRNA agent is 15-30 nucleotide pairs in length.

7. The iRNA agent of claim 1, comprising a modification that causes the iRNA agent to have increased stability in a biological sample.

8. The iRNA agent of claim 1, comprising a phosphorothioate or a 2′

-modified nucleotide.

9. The iRNA agent of claim 1, comprising at least one 5′

-uridine-adenine-3′

(5′

-ua-3′

) dinucleotide wherein the uridine is a 2′

-modified nucleotide;

at least one 5′

-uridine-guanine-3′

(5′

-ug-3′

) dinucleotide, wherein the 5′

-uridine is a 2′

-modified nucleotide;

at least one 5′

-cytidine-adenine-3′

(5′

-ca-3′

) dinucleotide, wherein the 5′

-cytidine is a 2′

-modified nucleotide;

or at least one 5′

-uridine-uridine-3′

(5′

-uu-3′

) dinucleotide, wherein the 5′

-uridine is a 2′

-modified nucleotide.

10. The iRNA agent of claim 1, wherein the 2′

-modification is selected from the group consisting of;

2′

-deoxy, 2′

-deoxy-2′

-fluoro, 2′

-O-methyl, 2′

-O-methoxyethyl (2′

-O-MOE), 2′

-O-aminopropyl (2′

-O-AP), 2′

-O-dimethylaminoethyl (2′

-O-DMAOE), 2′

-O-dimethylaminopropyl (2′

-O-DMAP), 2′

-O-dimethylaminoethyloxyethyl (2′

-O-DMAEOE), and 2′

-O—

N-methylacetamido (2′

-O-NMA).

11. The iRNA agent of claim 1, comprising a nucleotide overhang having 1 to 4 unpaired nucleotides.

12. The iRNA agent of claim 11, wherein the nucleotide overhang has 2 or 3 unpaired nucleotides.

13. The iRNA agent of claim 11, wherein the nucleotide overhang is at the 3′

-end of the antisense strand of the iRNA agent.

14. The iRNA agent of claim 1, comprising an epithelial receptor ligand.

15. The iRNA agent of claim 1, wherein the iRNA agent is targeted for uptake by cells of the lung.

16. A method of treating a human subject having a pathological process mediated at least in part by alpha-ENaC expression, wherein the iRNA agent comprises a sense strand wherein the sense strand comprises at least 15 contiguous nucleotides that differ by no more than 1, 2, or 3 nucleotides from the sense strand sequences any one of the agents provided in claim 1.

17. The method of claim 16, wherein the iRNA agent is administered in an amount sufficient to reduce the level of alpha-ENaC expression in a cell or tissue of the subject.

18. A pharmaceutical composition, comprising:

a) an iRNA agent of claim 1; and

b) a pharmaceutically acceptable carrier.

19. A method of treating a human subject afflicted with cystic fibrosis which method comprises administering to said subject a therapeutically effective amount of a iRNA agent of claim 1.

20. A method of treating a human subject afflicted with Liddles syndrome which method comprises administering to said patient a therapeutically effective amount of an iRNA agent of claim 1.

21. A method of treating and or preventing hypertension and or renal insufficiency in a human subject which method comprises administering to said patient a therapeutically effective amount of an iRNA agent of claim 1.

22. A method of treating and or preventing an electrolyte imbalance in a human subject which method comprises administering to said patient a therapeutically effective amount of an iRNA agent of claim 1.

2. An iRNA agent including a sense strand, wherein the sense strand comprises at least 15 contiguous nucleotides that differ by no more than 1, 2, or 3 nucleotides from the sense strand sequences of any one of the agents provided in Tables 1A-1D, agents numbered ND8285-ND10788, and an antisense strand wherein the antisense strand comprises at least 15 contiguous nucleotides of the antisense sequences of any one of the agents provided in Tables 1A-1D, agents numbered ND8285-ND10788, and wherein the iRNA agent reduces the expression of alpha-ENaC by more than 20%, 30%, 40%, 50%, 60%, 70%, or 80% compared to cells which have not been incubated with the iRNA agent.

3. An iRNA agent comprising a sense strand and an antisense strand each comprising a sequence of at least 16, 17 or 18 nucleotides which is essentially identical to one of the sequences of any one of the agents provided in Tables 1A-1D, agents numbered ND8285-ND10788, except that not more than 1, 2 or 3 nucleotides per strand, respectively, have been substituted by other nucleotides, while essentially retaining the ability to reduce the amount of alpha-ENaC expression in cells.