MUCIN 5B (MUC5B) IRNA COMPOSITIONS AND METHODS OF USE THEREOF
The present invention relates to RNAi agents, e.g., dsRNA agents, targeting the mucin 5B (MUC5B) gene. The invention also relates to methods of using such RNAi agents to inhibit expression of a MUC5B gene and to methods of treating or preventing a MUC5B-associated disease, such as a lung disease, e.g., cystic fibrosis, chronic obstructive pulmonary disease (COPD), and/or pulmonary fibrosis, e.g., idiopathic pulmonary fibrosis (IPF), in a subject.
1 . A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of Mucin 5B (MUC5B) in a cell,
(a) wherein the dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand comprises a nucleotide sequence comprising at least 15 contiguous nucleotides, with 0, 1, 2, or 3 mismatches, of a portion of the nucleotide sequence of SEQ ID NO:1, or a nucleotide sequence having at least 90% nucleotide sequence identity to a portion of the nucleotide sequence of SEQ ID NO:1, and the antisense strand comprises a nucleotide sequence comprising at least 15 contiguous nucleotides, with 0, 1, 2, or 3 mismatches, of the corresponding portion of the nucleotide sequence of SEQ ID NO:6, or a nucleotide sequence having at least 90% nucleotide sequence identity to a portion of the nucleotide sequence of SEQ ID NO:6; and
wherein the sense strand or the antisense strand is conjugated to one or more lipophilic moieties; or
(b) wherein the antisense strand comprises a region complementary to part of an mRNA encoding a MUC5B gene (SEQ ID NO:1), wherein each strand independently is 14 to 30 nucleotides in length; and wherein the sense strand or the antisense strand is conjugated to one or more lipophilic moieties; or
(c) wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any one of the antisense nucleotide sequences in any one of Tables 2-7, 9, and 10, wherein each strand independently is 14 to 30 nucleotides in length; and wherein the sense strand or the antisense strand is conjugated to one or more lipophilic moieties.
2 . (canceled)
3 . (canceled)
4 . The dsRNA agent of claim 1 , wherein the sense strand or the antisense strand is a sense strand or an antisense strand selected from the group consisting of any of the sense strands and antisense strands in any one of Tables 2-7, 9, and 10.
5 .- 10 . (canceled)
11 . The dsRNA agent of claim 1 , wherein the dsRNA agent comprises at least one modified nucleotide.
12 . (canceled)
13 . (canceled)
14 . The dsRNA agent of claim 11 , wherein at least one of the modified nucleotides is selected from the group a deoxy-nucleotide, a 3′-terminal deoxy-thymine (dT) nucleotide, a 2′-O-methyl modified nucleotide, a 2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an unlocked nucleotide, a conformationally restricted nucleotide, a constrained ethyl nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-O-allyl-modified nucleotide, 2′-C-alkyl-modified nucleotide, a 2′-methoxyethyl modified nucleotide, a 2′-O-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, a non-natural base comprising nucleotide, a tetrahydropyran modified nucleotide, a 1,5-anhydrohexitol modified nucleotide, a cyclohexenyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, a nucleotide comprising a 5′-methylphosphonate group, a nucleotide comprising a 5′ phosphate or 5′ phosphate mimic, a nucleotide comprising vinyl phosphonate, a nucleotide comprising adenosine-glycol nucleic acid (GNA), a nucleotide comprising thymidine-glycol nucleic acid (GNA)S-Isomer, a nucleotide comprising 2-hydroxymethyl-tetrahydrofurane-5-phosphate, a nucleotide comprising 2′-deoxythymidine-3′phosphate, a nucleotide comprising 2′-deoxyguanosine-3′-phosphate, a 2′-O hexadecyl nucleotide, a nucleotide comprising a 2′-phosphate, a cytidine-2′-phosphate nucleotide, a guanosine-2′-phosphate nucleotide, a 2′-O-hexadecyl-cytidine-3′-phosphate nucleotide, a 2′-O-hexadecyl-adenosine-3′-phosphate nucleotide, a 2′-O-hexadecyl-guanosine-3′-phosphate nucleotide, a 2′-O-hexadecyl-uridine-3′-phosphate nucleotide, a a 5′-vinyl phosphonate (VP), a 2′-deoxyadenosine-3′-phosphate nucleotide, a 2′-deoxycytidine-3′-phosphate nucleotide, a 2′-deoxyguanosine-3′-phosphate nucleotide, a 2′-deoxythymidine-3′-phosphate nucleotide, a 2′-deoxyuridine nucleotide, and a terminal nucleotide linked to a cholesteryl derivative and a dodecanoic acid bisdecylamide group; and combinations thereof.
15 .- 17 . (canceled)
18 . The dsRNA agent of claim 14 , further comprising at least one phosphorothioate internucleotide linkage.
19 . (canceled)
20 . The dsRNA agent of claim 1 , wherein each strand is no more than 30 nucleotides in length.
21 .- 31 . (canceled)
32 . The dsRNA agent of claim 1 , wherein one or more lipophilic moieties are conjugated to one or more internal positions on at least one strand.
33 .- 41 . (canceled)
42 . The dsRNA agent of claim 1 , wherein the one or more lipophilic moieties are conjugated to one or more of the internal positions selected from the group consisting of positions 4-8 and 13-18 on the sense strand, and positions 6-10 and on the antisense strand, counting from the 5′-end of each strand.
43 .- 51 . (canceled)
52 . The dsRNA agent of claim 1 , wherein the lipophilic moiety contains a saturated or unsaturated C4-C30 hydrocarbon chain, and an optional functional group selected from the group consisting of hydroxyl, amine, carboxylic acid, sulfonate, phosphate, thiol, azide, and alkyne.
53 .- 68 . (canceled)
69 . The dsRNA agent of claim 1 , further comprising a phosphate or phosphate mimic at the 5′-end of the antisense strand.
70 .- 72 . (canceled)
73 . An isolated cell containing the dsRNA agent of claim 1 .
74 . A pharmaceutical composition for inhibiting expression of a MUC5B gene, comprising the dsRNA agent of claim 1 .
75 . (canceled)
76 . A device for oral inhalative administration comprising the dsRNA agent of claim 1 .
77 . (canceled)
78 . An in vitro method of inhibiting expression of a MUC5B gene in a cell, the method comprising:
(a) contacting the cell with the dsRNA agent of claim 1 ; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the MUC5B gene, thereby inhibiting expression of the MUC5B gene in the cell.
79 . (canceled)
80 . (canceled)
81 . The method of claim 76 , wherein the expression of the MUC5B gene is inhibited by at least 50%.
82 . A method of treating a subject having a Mucin 5B- (MUC5B-) associated disease or a subject at risk of developing a MUC5B-associated disease, comprising administering to the subject a therapeutically effective amount of the dsRNA agent of claim 1 , thereby treating said subject.
83 . The method of claim 82 , wherein the subject is a human.
84 . The method of claim 82 , wherein the MUC5B-associated disease is pulmonary fibrosis, cystic fibrosis, or chronic obstructive pulmonary disease (COPD).
85 . (canceled)
86 . (canceled)
87 . The method of claim 82 , wherein the dsRNA agent is administered to the subject at a dose of about 0.01 mg/kg to about 50 mg/kg.
88 .- 90 . (canceled)
91 . The method of claim 82 , further comprising administering to the subject an additional agent or a therapy suitable for treatment or prevention of a MUC5B-associated disorder.
92 .- 95 . (canceled)