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 for inhibiting the expression of alpha-ENaC, wherein the iRNA agent comprises a sense strand and an antisense strand, wherein: the antisense strand comprises a nucleotide sequence of nucleotides 2-18 of SEQ ID NO: 400, SEQ ID NO: 506, or SEQ ID NO: 1120, and the sense strand is complementary to the antisense strand.
2. The composition of claim 1 , wherein: the sense strand of the iRNA agent comprises a nucleotide sequence of nucleotides 2-18 of SEQ ID NO: 399, SEQ ID NO: 505 or SEQ ID NO: 1119.
3. The composition of claim 1 , wherein the antisense strand and the sense strand of the iRNA agent are each 19 to 23 nucleotides in length.
4. The composition of claim 1 , wherein the iRNA agent comprises at least one modified nucleotide and/or at least one phosphate linker modification.
5. The composition of claim 4 , wherein the iRNA agent comprises one or more 2′-modified nucleotides and/or one or more phosphorothioates.
6. The composition of claim 5 , wherein the one or more 2′-modified nucleotides are independently 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).
7. The composition of claim 6 , wherein the iRNA agent comprises one or more chemically modified nucleotides and one or more phosphate linker modifications.
8. The composition of claim 4 , wherein the sense strand and/or the antisense strand contains a 3′ overhang.
9. The composition of claim 4 , wherein the sense strand and/or the antisense strand contains a 5′ overhang.
10. The composition of claim 4 , wherein the iRNA agent comprises at least one blunt end.
11. The composition of claim 4 , further comprising a buffer, diluent, penetration enhancer, carrier compound, and/or pharmaceutically acceptable carrier or excipient.
12. The composition of claim 4 , wherein the composition is presented in unit dosage form.
13. 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, epithelial receptor ligand, polycationic, peptide, polyamine, peptide mimic, and/or transferrin.
14. The composition of claim 4 , further comprising one or more known agents effective in treatment of ENaC-related disorders.
15. The composition of claim 14 , wherein the known agent is selected from the group consisting of: anti-inflammatory drug, bronchodilatory drug, antihistamine, anti-tussive drug, antibiotic, DNase drug substance, epithelial sodium channel blocker.
16. The composition of claim 4 , wherein the iRNA agent comprises:
a) an antisense strand comprising a nucleotide sequence of nucleotides 1-19 of SEQ ID NO: 400 and a sense strand comprising a nucleotide sequence of nucleotides 1-19 of SEQ ID NO: 399,
b) an antisense strand comprising a nucleotide sequence of nucleotides 1-19 of SEQ ID NO: 506 and a sense strand comprising a nucleotide sequence of nucleotides 1-19 of SEQ ID NO: 505, or
c) an antisense strand comprising a nucleotide sequence of nucleotides 1-19 of SEQ ID NO: 1120 and a sense strand comprising a nucleotide sequence of nucleotides 1-19 of SEQ ID NO: 1119.
17. 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 4 .
18. The method of claim 17 , 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.
19. The method of claim 17 , wherein the composition is administered via inhalation/intranasal administration or systemically or subcutaneously.
20. A method of inhibiting the expression of an alpha-ENaC gene in a cell, the method comprising administering to the cell an effective amount of a composition of claim 4 .