RNAi agents for inhibiting expression of Beta-ENaC, compositions thereof, and methods of use
Described are RNAi agents, compositions that include RNAi agents, and methods for inhibition of a beta-ENaC (SCNN1B) gene. The beta-ENaC RNAi agents and RNAi agent conjugates disclosed herein inhibit the expression of a beta-ENaC gene. Pharmaceutical compositions that include one or more beta-ENaC RNAi agents, optionally with one or more additional therapeutics, are also described. Delivery of the described beta-ENaC RNAi agents to epithelial cells, such as pulmonary epithelial cells, in vivo, provides for inhibition of beta-ENaC gene expression and a reduction in ENaC activity, which can provide a therapeutic benefit to subjects, including human subjects, for the treatment of various diseases including chronic obstructive pulmonary disease (COPD).
1 . An RNAi agent for inhibiting expression of a beta-ENaC gene, comprising:
an antisense strand comprising the nucleotide sequence (5′→3′) cPrpusGfsusUfgAfagaugUfaAfcAfgUfuGfsc (SEQ ID NO: 138); and
a sense strand comprising a nucleotide sequence that is at least partially complementary to the antisense strand;
wherein a represents 2′-O-methyl adenosine, c represents 2′-O-methyl cytidine, g represents 2′-O-methyl guanosine, and u represents 2′-O-methyl uridine; Af represents 2′-fluoro adenosine, Cf represents 2′-fluoro cytidine, Gf represents 2′-fluoro guanosine, and Uf represents 2′-fluoro uridine; cPrpu represents a 5′-cyclopropyl phosphonate-2′-O-methyl uridine; s represents a phosphorothioate linkage; and wherein all or substantially all of the nucleotides on the sense strand are modified nucleotides.
2 . The RNAi agent of claim 1 , wherein the sense strand comprises the nucleotide sequence (5′→3′) gscaacuguUfAfCfaucuucaaca (SEQ ID NO: 239)
wherein a represents 2′-O-methyl adenosine, c represents 2′-O-methyl cytidine, g represents 2′-O-methyl guanosine, and u represents 2′-O-methyl uridine; Af represents 2′-fluoro adenosine, Cf represents 2′-fluoro cytidine, Gf represents 2′-fluoro guanosine, and Uf represents 2′-fluoro uridine; s represents a phosphorothioate linkage.
3 . The RNAi agent of claim 1 , wherein the sense strand comprises one or two inverted abasic residues.
4 . The RNAi agent of claim 1 , wherein the RNAi agent comprises of a sense strand and an antisense strand that form a duplex having the structure of AD07099 (SEQ ID NO: 138 and SEQ ID NO: 177.
5 . The RNAi agent of claim 1 , wherein the RNAi agent is linked to a targeting ligand.
6 . The RNAi agent of claim 5 , wherein the targeting ligand comprises the structure:
or a pharmaceutically acceptable salt thereof, wherein indicates the point of connection to the RNAi agent.
7 . The RNAi agent of claim 5 , wherein the targeting ligand has a structure selected from the group consisting of:
wherein indicates the point of connection to the RNAi agent.
8 . The RNAi agent of claim 1 , wherein
the antisense strand comprises the structure (5′→3′):
cPrpusGfsusUfgAfagaugUfaAfcAfgUfuGfsc (SEQ ID NO: 138); and
the sense strand comprises the structure (5′→3′):
(TriSM6.1-avb6-TA14) gscaacuguUfAfCfaucuucaacas (invAb) (SEQ ID NO: 187);
wherein a represents 2′-O-methyl adenosine, c represents 2′-O-methyl cytidine, g represents 2′-O-methyl guanosine, and u represents 2′-O-methyl uridine; Af represents 2′-fluoro adenosine, Cf represents 2′-fluoro cytidine, Gf represents 2′-fluoro guanosine, and Uf represents 2′-fluoro uridine; cPrpu represents a 5′-cyclopropyl phosphonate-2′-O-methyl uridine; s represents a phosphorothioate linkage; (invAb) represents an inverted abasic residue; and (TriSM6.1-avb6-TA14) represents:
pharmaceutically acceptable salt thereof, wherein represents the remainder of the RNAi agent.
9 . A method for inhibiting expression of a beta-ENaC gene in a cell, the method comprising introducing into a cell an effective amount of an RNAi agent of claim 1 .
10 . A method of treating one or more symptoms or diseases associated with enhanced or elevated ENaC activity levels, the method comprising administering to a human subject in need thereof a therapeutically effective amount of the RNAi agent of claim 1 .
11 . The method of claim 10 , wherein the disease is a respiratory disease.
12 . The method of claim 11 , wherein the respiratory disease is cystic fibrosis, chronic bronchitis, non-cystic fibrosis bronchiectasis, chronic obstructive pulmonary disease (COPD), asthma, respiratory tract infections, primary ciliary dyskinesia, or lung carcinoma cystic fibrosis.
13 . The method of claim 10 , wherein the RNAi agent is administered at a deposited dose of about 0.01 mg/kg to about 5.0 mg/kg of body weight of the subject.