RNAi inhibition of alpha-ENaC expression

• US 7,943,592 B2
• Filed: 04/30/2010
• Issued: 05/17/2011
• Est. Priority Date: 06/15/2007
• Status: Active Grant

• Current Assignee: Arrowhead Research Corporation
• Original Assignee: Novartis Ag
• Inventors: Geick, Anke, Tan, Pamela, Van Heeke, Gino, Danahay, Henry Luke, Vornlocher, Hans Peter, Hickman, Emma
• Primary Examiner(s): MCGARRY, SEAN
• Application Number:
  US12/771,470
• Publication Number:
  US 20100210513A1
• Time in Patent Office: 382 Days
• Field of Search: None
• US Class Current: 514/44.00A
• 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;

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

224;

or

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

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

528;

or

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

1281, and the sequence of the second strand is the sequence of SEQ ID NO;

1282.

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).

9. 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 strand and the second strand are the sequences of SEQ ID NOs:

223 and 224, respectively.

15. The composition of claim 6, wherein the iRNA agent comprises a phosphorothioate or a 2′

-modified nucleotide.

16. The composition of claim 6, 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.

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

527 and 528, respectively.

17. The composition of claim 7, wherein the iRNA agent comprises a phosphorothioate or a 2′

-modified nucleotide.

18. The composition of claim 7, 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.

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

1281 and 1282, respectively.

19. The composition of claim 8, wherein the iRNA agent comprises a phosphorothioate or a 2′

-modified nucleotide.

20. The composition of claim 8, 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.

10. 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 is the sequence of SEQ ID NO:

223, SEQ ID NO;

527, or SEQ ID NO;

1281, and

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

224, SEQ ID NO;

528, or SEQ ID NO;

1282,

the composition further comprising an epithelial receptor ligand.

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

12. 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;

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

224;

or

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

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

528;

or

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

1281, and the sequence of the second strand is the sequence of SEQ ID NO;

1282.

13. The method of claim 12, 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.

14. The method of claim 12, wherein the human subject has a pathological state selected from the group of:

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