RNAi Inhibition of Alpha-ENaC Expression

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.

1 . A composition comprising an iRNA agent for inhibiting the expression of an alpha-ENaC gene, 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: 980, SEQ ID NO: 1298, or SEQ ID NO: 1368. 
     
     
         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: 979, SEQ ID NO: 1297, or SEQ ID NO: 1367. 
     
     
         3 . The composition of  claim 1 , wherein the antisense strand of the iRNA agent 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 2 , wherein the iRNA agent comprises at least on modified nucleotide and/or at least one phosphate linker modification. 
     
     
         6 . The composition of  claim 4 , wherein the iRNA agent comprises one or more 2′-modified nucleotides and/or one or more phosphorothioates. 
     
     
         7 . The composition of  claim 5 , wherein the iRNA agent comprises one or more 2′-modified nucleotides and/or one or more phosphorothioates. 
     
     
         8 . The composition of  claim 6 , 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). 
     
     
         9 . The composition of  claim 7 , 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). 
     
     
         10 . The composition of  claim 4 , wherein the sense strand and/or the antisense strand contains a 3′ overhang. 
     
     
         11 . The composition of  claim 4 , wherein the sense strand and/or the antisense strand contains a 5′ overhang. 
     
     
         12 . The composition of  claim 4 , wherein the iRNA agent comprises at least one blunt end. 
     
     
         13 . The composition of  claim 4 , further comprising a buffer, diluent, penetration enhancer, carrier compound, and/or pharmaceutically acceptable carrier or excipient. 
     
     
         14 . The composition of  claim 4 , wherein the composition is presented in unit dosage form. 
     
     
         15 . 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. 
     
     
         16 . The composition of  claim 4 , further comprising one or more known agents effective in treatment of ENaC-related disorders. 
     
     
         17 . The composition of  claim 16 , 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. 
     
     
         18 . The composition of  claim 5 , wherein the iRNA agent comprises:
 a) an antisense strand comprising a nucleotide sequence of nucleotides 1-19 of SEQ ID NO: 980, and a sense strand comprising a nucleotide sequence of nucleotides 1-19 of SEQ ID NO: 979,   b) an antisense strand comprising a nucleotide sequence of nucleotides 1-19 of SEQ ID NO: 1298 and a sense strand comprising a nucleotide sequence of nucleotides 1-19 of SEQ ID NO: 1297, or   c) an antisense strand comprising a nucleotide sequence of nucleotides 1-19 of SEQ ID NO: 1368, and a sense strand comprising a nucleotide sequence of nucleotides 1-19 of SEQ ID NO: 1367.   
     
     
         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 4 . 
     
     
         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. 
     
     
         21 . The method of  claim 19 , wherein the composition is administered via inhalation/intranasal administration or systemically or subcutaneously. 
     
     
         22 . 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 . 
     
     
         23 . A composition comprising an iRNA agent comprising a sense strand and an antisense strand, wherein: the antisense strand comprises a nucleotide sequence of nucleotides 2-18 of any of the antisense strands Table 1A, Table 1B, Table 1C, or Table 1D.