RNAi inhibition of alpha-ENaC expression
View Patent ↗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.
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: 1511, and the sequence of the second strand is the sequence of SEQ ID NO: 1512; or
b. the sequence of the first strand is the sequence of SEQ ID NO: 1637, and sequence of the second strand is the sequence of SEQ ID NO: 1638.
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).
6. 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.
7. A composition comprising an iRNA agent to alpha-ENaC, wherein the iRNA agent comprises a first strand and a second strand,
wherein the first strand comprises the sequence of SEQ ID NO: 1511, or SEQ ID NO: 1637, and
wherein the second strand comprises the sequence of SEQ ID NO: 1512, or SEQ ID NO: 1638,
the composition further comprising an epithelial receptor ligand.
8. The composition of claim 7 , wherein the epithelial receptor ligand is (i) transferrin or (ii) folic acid.
9. 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 comprises the sequence of SEQ ID NO: 1511, and the sequence of the second strand comprises the sequence of SEQ ID NO: 1512; or
b. the sequence of the first strand comprises the sequence of SEQ ID NO: 1637, and sequence of the second strand comprises the sequence of SEQ ID NO: 1638.
10. The method of claim 9 , 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.
11. The method of claim 9 , 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 electrolyte imbalance.
12. 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: 1511, and the sequence of the second strand comprises the sequence of SEQ ID NO: 1512; or
b. the sequence of the first strand comprises the sequence of SEQ ID NO: 1637, and the sequence of the second strand comprises the sequence of SEQ ID NO: 1638.
13. The composition of claim 12 , wherein the second strand is 30 or fewer nucleotides in length, and wherein the first strand and the antisense strand form a duplex region 15 to 30 nucleotide pairs in length.
14. The composition of claim 12 , wherein the second strand and the first strand are each 19 to 23 nucleotides in length.
15. The composition of claim 12 , wherein the iRNA agent comprises a blunt end.
16. The composition of claim 12 , wherein the iRNA agent comprises a nucleotide overhang having 1 to 4 unpaired nucleotides.
17. The composition of claim 12 , wherein the iRNA agent comprises a nucleotide overhang at the 3′-end of the antisense strand of the iRNA agent.
18. The composition of claim 12 , wherein the iRNA agent comprises a modification that causes the iRNA agent to have increased stability in a biological sample.
19. The composition of claim 12 , wherein the iRNA agent comprises a phosphorothioate or a 2′-modified nucleotide.
20. The composition of claim 12 , 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.
21. The composition of claim 12 , 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).
22. The composition of claim 1 , wherein the iRNA agent is not modified.
23. The composition of claim 12 , wherein the iRNA agent is not modified.