RNAi inhibition of alpha-ENaC expression

• US 8,119,612 B2
• Filed: 04/30/2010
• Issued: 02/21/2012
• Est. Priority Date: 06/15/2007
• Status: Active Grant

• Current Assignee: Arrowhead Pharmaceuticals, Inc.
• Original Assignee: Novartis Ag
• Inventors: Van Heeke, Gino, Hickman, Emma, Danahay, Henry Luke, Tan, Pamela, Geick, Anke, Vornlocher, Hans Peter
• Primary Examiner(s): McGarry, Sean
• Application Number:
  US12/771,566
• Publication Number:
  US 20100216972A1
• Time in Patent Office: 662 Days
• Field of Search: None
• US Class Current: 514/44.A
• 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. A composition comprising an iRNA agent comprising a first strand and a second strand, wherein:

a. the sequence of the first strand is the sequence of SEQ ID NO;

145, and the sequence of the second strand is the sequence of SEQ ID NO;

146;

or

b. the sequence of the first strand is the sequence of SEQ ID NO;

57, and the sequence of the second strand is the sequence of SEQ ID NO;

58.

2. The composition of claim 1, wherein the iRNA agent comprises a modification that causes the iRNA agent to have increased stability in a biological sample.

3. The composition of claim 1, wherein the iRNA agent comprises a phosphorothioate or a 2′

-modified nucleotide.

4. The composition of claim 1, wherein the iRNA agent comprises:

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.

5. The composition of claim 1, wherein the iRNA agent comprises a 2′

-modification 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).

8. The composition of claim 1, wherein the iRNA agent is ligated to one or more diagnostic compound, reporter group, cross-linking agent, nuclease-resistance conferring moiety, natural or unusual nucleobase, lipophilic molecule, cholesterol, lipid, lectin, steroid, uvaol, hecigenin, diosgenin, terpene, triterpene, sarsasapogenin, Friedelin, epifriedelanol-derivatized lithocholic acid, vitamin, carbohydrate, dextran, pullulan, chitin, chitosan, synthetic carbohydrate, Oligo Lactate 15-mer, natural polymer, low- or medium-molecular weight polymer, inulin, cyclodextrin, hyaluronic acid, protein, protein-binding agent, integrin-targeting molecule, polycationic, peptide, polyamine, peptide mimic, and/or transferrin.

6. A composition comprising an iRNA agent comprising a first strand and a second strand, wherein the sequences of the first and second strand are the sequences of SEQ ID NOs:

145 and 146, respectively.

7. A composition comprising an iRNA agent comprising a first strand and a second strand, wherein the sequences of the first and second strand are the sequences of SEQ ID NOs:

57 and 58, respectively.

9. A composition comprising an iRNA agent to alpha-ENaC, wherein the iRNA agent comprises a first strand and a second strand, wherein the sequence of the first strand comprises the sequence of SEQ ID NO:

145, or SEQ ID NO;

57, and

wherein the sequence of the second strand comprises the sequence of SEQ ID NO;

146, or SEQ ID NO;

58,

the composition further comprising an epithelial receptor ligand.

10. The composition of claim 9, wherein the epithelial receptor ligand is (i) transferrin or (ii) folic acid.

11. A method of treating a human subject having a pathological state mediated at least in part by alpha-ENaC expression, the method comprising the step of administering to the subject a therapeutically effective amount of a composition comprising an iRNA agent comprising a first strand and a second strand, wherein:

a. the sequence of the first strand is the sequence of SEQ ID NO;

145, and the sequence of the second strand is the sequence of SEQ ID NO;

146;

or

b. the sequence of the first strand is the sequence of SEQ ID NO;

57, and the sequence of the second strand is the sequence of SEQ ID NO;

58.

12. The method of claim 11, wherein the composition is administered in an amount sufficient to reduce the level of alpha-ENaC expression in a cell or tissue of a human subject.

13. The use method of claim 11, wherein the human subject has a pathological state selected from:

cystic fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease (COPD), asthma, respiratory tract infections, lung carcinoma, Liddles syndrome, hypertension, renal insufficiency, and/or electrolyte imbalance.

14. A composition comprising an iRNA agent comprising a first strand and a second strand, wherein:

a. the sequence of the first strand comprises the sequence of SEQ ID NO;

145, and the sequence of the second strand comprises the sequence of SEQ ID NO;

146;

or

b. the sequence of the first strand comprises the sequence of SEQ ID NO;

57, and the sequence of the second strand comprises the sequence of SEQ ID NO;

58.

15. The composition of claim 14, wherein the second strand is 30 or fewer nucleotides in length, and wherein the first strand and the second strand form a duplex region 15 to 30 nucleotide pairs in length.

16. The composition of claim 14, wherein the second strand and the first strand are each 19 to 23 nucleotides in length.

17. The composition of claim 14, wherein the iRNA agent comprises a blunt end.

18. The composition of claim 14, wherein the iRNA agent comprises a nucleotide overhang having 1 to 4 unpaired nucleotides.

19. The composition of claim 14, wherein the iRNA agent comprises a nucleotide overhang at the 3′

-end of the antisense strand of the iRNA agent.

20. The composition of claim 14, wherein the iRNA agent comprises a modification that causes the iRNA agent to have increased stability in a biological sample.

21. The composition of claim 14, wherein the iRNA agent comprises a phosphorothioate or a 2′

-modified nucleotide.

22. The composition of claim 14, wherein the iRNA agent comprises:

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.

23. The composition of claim 14, wherein the iRNA agent comprises a 2′

-modification 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).