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 sense strand and an antisense strand, wherein:
a. the antisense strand comprises the nucleotide sequence of nucleotides 1-19 of SEQ ID NO: 980.
2. The composition of claim 1 , wherein the sense strand comprises the nucleotide sequence of nucleotides 1-9 of SEQ ID NO: 979.
3. The composition of claim 1 , wherein the antisense strand is 30 or fewer nucleotides in length, and wherein the sense strand and the antisense strand form a duplex region 15 to 30 nucleotide pairs in length.
4. The composition of claim 1 , wherein the antisense strand and the sense strand are each 19 to 23 nucleotides in length.
5. 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.
6. The composition of claim 1 , wherein the iRNA agent comprises at least one phosphorothioate or 2′-modified nucleotide.
7. The composition of claim 1 , wherein the iRNA agent comprises: least one 5′-uridine-adenine-3′ (5′-ua-3′) dinucleotide, wherein the uridine is a 2′-modified nucleotide; at least one 5′-uridineguanine-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; and/or at least one 5′-uridine-uridine-3′ (5′-uu-3′) dinucleotide, wherein the 5′-uridine is a 2′modified nucleotide.
8. 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-dimethylarninopropyl (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 comprises a blunt end.
10. The composition of claim 1 , wherein the iRNA agent comprises a nucleotide overhang having 1 to 4 unpaired nucleotides.
11. The composition of claim 1 , wherein the iRNA agent comprises a nucleotide overhang at the 3′-end of the antisense strand of the iRNA agent.
12. A composition comprising an iRNA agent comprising a sense strand and an antisense strand, wherein the antisense strand comprises the nucleotide sequence of nucleotides 1-19 of SEQ ID NO: 1368 wherein the cytidines at positions 15 and 17 are 2′-O-methylcytidines.
13. The composition of claim 12 wherein the sense strand comprises nucleotides 1-19 of SEQ ID NO 1367.
14. 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.
15. 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 of claim 1 .
16. The method of claim 15 , wherein the pathological state is 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.
17. A composition comprising an iRNA agent to alpha-ENaC, wherein the iRNA agent comprises a sense strand and an antisense strand, wherein the antisense strand comprises the nucleotide sequence of nucietoides 1-19 of SEQ ID NO: 980, the composition further comprising an epithelial receptor ligand.
18. The composition of claim 17 , wherein the sense strand comprises the nucleotide sequence of nucleotides 1-19 of SEQ ID NO 979.
19. 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 of claim 18 .
20. The method of claim 19 , wherein the pathological state is 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.