Transmembrane serine protease 2 (TMPRSS2) iRNA compositions and methods of use thereof
The present invention relates to RNAi agents, e.g., dsRNA agents, targeting the transmembrane serine protein 2 (TMPRSS2) gene. The invention also relates to methods of using such RNAi agents to inhibit expression of a TMPRSS2 gene and to methods of treating or preventing a TMPRSS2-associated disease, e.g., COVID-19, in a subject.
1 . A double stranded ribonucleic acid (dsRNA) agent, or a salt thereof, for inhibiting expression of transmembrane serine protease 2 (TMPRSS2) in a cell,
wherein the dsRNA agent, or a salt thereof, comprises a sense strand and an antisense strand forming a double stranded region,
wherein the antisense strand comprises at least 15 contiguous nucleotides of the nucleotide sequence 5′-UAGUAAUGAUAGGUAUCUGGAAU-3′ of SEQ ID NO: 323; and
wherein the sense strand or the antisense strand is conjugated to one or more lipophilic moieties.
2 . The dsRNA agent, or a salt thereof, of claim 1 , wherein the antisense strand comprises at least 17 contiguous nucleotides of the nucleotide sequence 5′-UAGUAAUGAUAGGUAUCUGGAAU-3′ of SEQ ID NO:323.
3 . The dsRNA agent, or a salt thereof, of claim 1 , wherein at least one nucleotide comprises a nucleotide modification.
4 . The dsRNA agent, or a salt thereof, of claim 3 , wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a nucleotide modification.
5 . The dsRNA agent, or a salt thereof, of any claim 3 , wherein at least one of the nucleotide modifications is selected from the group a deoxy-nucleotide modification, a 3′-terminal deoxy-thymine (dT) nucleotide modification, a 2′-O-methyl nucleotide modification, a 2′-fluoro nucleotide modification, a 2′-deoxy nucleotide modification, a locked nucleotide modification, an unlocked nucleotide modification, a conformationally restricted nucleotide modification, a constrained ethyl nucleotide modification, an abasic nucleotide modification, a 2′-amino nucleotide modification, a 2′-O-allyl nucleotide modification, 2′-C-alkyl-modified nucleotide modification, a 2′-methoxyethyl nucleotide modification, a 2′-O-alkyl nucleotide modification, a morpholino nucleotide modification, a phosphoramidate modification, a non-natural base comprising nucleotide modification, a tetrahydropyran nucleotide modification, a 1,5-anhydrohexitol nucleotide modification, a cyclohexenyl nucleotide modification, a nucleotide comprising a 5′-phosphorothioate group modification, a nucleotide comprising a 5′-methylphosphonate group modification, a nucleotide comprising a 5′ phosphate or 5′ phosphate mimic modification, a nucleotide comprising vinyl phosphonate modification, a nucleotide comprising adenosine-glycol nucleic acid (GNA) modification, a nucleotide comprising thymidine-glycol nucleic acid (GNA)S-Isomer modification, a nucleotide comprising 2-hydroxymethyl-tetrahydrofurane-5-phosphate modification, a nucleotide comprising 2′-deoxythymidine-3′phosphate modification, a nucleotide comprising 2′-deoxyguanosine-3′-phosphate modification, a 2′-O hexadecyl nucleotide modification, a nucleotide comprising a 2′-phosphate modification, a cytidine-2-phosphate nucleotide modification, a guanosine-2-phosphate nucleotide modification, a 2′-O-hexadecyl-cytidine-3′-phosphate nucleotide modification, a 2′-O-hexadecyl-adenosine-3′-phosphate nucleotide modification, a 2′-O-hexadecyl-guanosine-3′-phosphate nucleotide modification, a 2′-O-hexadecyl-uridine-3′-phosphate nucleotide modification, a 5′-vinyl phosphonate (VP) modification, a 2-deoxyadenosine-3-phosphate nucleotide modification, a 2′-deoxycytidine-3-phosphate nucleotide modification, a 2-deoxyguanosine-3-phosphate nucleotide modification, a 2-deoxythymidine-3-phosphate nucleotide modification, a 2-deoxyuridine nucleotide modification, and a terminal nucleotide linked to a cholesteryl derivative and a dodecanoic acid bisdecylamide group modification; and combinations thereof.
6 . The dsRNA agent, or a salt thereof, of claim 5 , further comprising at least one phosphorothioate internucleotide linkage.
7 . The dsRNA agent, or a salt thereof, of claim 1 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide.
8 . The dsRNA agent, or a salt thereof, of claim 1 , wherein each strand is 19-30 nucleotides in length.
9 . The dsRNA agent, or a salt thereof, of claim 1 , wherein one or more lipophilic moieties are conjugated to one or more internal positions on at least one strand.
10 . The dsRNA agent, or a salt thereof, 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 15-18 on the antisense strand, counting from the 5′-end of each strand.
11 . The dsRNA agent, or a salt thereof, of claim 1 , wherein the lipophilic moiety is an aliphatic, alicyclic, or polyalicyclic compound.
12 . The dsRNA agent, or a salt thereof, of claim 11 , wherein the lipophilic moiety is selected from the group consisting of lipid, cholesterol, retinoic acid, cholic acid, adamantane acetic acid, 1-pyrene butyric acid, dihydrotestosterone, 1,3-bis-O(hexadecyl)glycerol, geranyloxyhexyanol, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecyl group, palmitic acid, myristic acid, O3-(oleoyl) lithocholic acid, 03-(oleoyl) cholenic acid, dimethoxytrityl, or phenoxazine.
13 . The dsRNA agent, or a salt thereof, of claim 12 , 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.
14 . The dsRNA agent, or a salt thereof, of claim 13 , wherein the lipophilic moiety contains a saturated or unsaturated C6-C18 hydrocarbon chain.
15 . The dsRNA agent, or a salt thereof, of claim 1 , further comprising a phosphate or phosphate mimic at the 5′-end of the antisense strand.
16 . The dsRNA agent, or a salt thereof, of claim 1 , wherein the base pair at the 1 position of the 5′-end of the antisense strand of the duplex is an AU base pair.
17 . An isolated cell containing the dsRNA agent, or a salt thereof, of claim 1 .
18 . A pharmaceutical composition for inhibiting expression of a TMPRSS2 gene, comprising the dsRNA agent, or a salt thereof, of any one of claim 1 .
19 . A double stranded ribonucleic acid (dsRNA) agent, or a salt thereof, for inhibiting expression of transmembrane serine protease 2 (TMPRSS2) in a cell,
wherein the dsRNA agent, or a salt thereof, comprises a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand comprises at least 15 contiguous nucleotides of the nucleotide sequence 5′-UCCAGAUACCUAUCAUUACUA-3′ of SEQ ID NO:23 and the antisense strand comprises at least 15 contiguous nucleotides of the nucleotide sequence 5′-UAGUAAUGAUAGGUAUCUGGAAU-3′ of SEQ ID NO:323; and
wherein the sense strand or the antisense strand is conjugated to one or more lipophilic moieties.
20 . The dsRNA agent, or a salt thereof, of claim 1 , wherein the antisense strand comprises at least 19 contiguous nucleotides of the nucleotide sequence 5′-UAGUAAUGAUAGGUAUCUGGAAU-3′ of SEQ ID NO:323.
21 . The dsRNA agent, or a salt thereof, of claim 1 , wherein the antisense strand comprises the nucleotide sequence 5′-UAGUAAUGAUAGGUAUCUGGAAU-3′ of SEQ ID NO: 323.
22 . The dsRNA agent, or a salt thereof, of claim 1 , wherein the sense strand comprises the nucleotide sequence 5′-UCCAGAUACCUAUCAUUACUA-3′ of SEQ ID NO:23 and the antisense strand comprises the nucleotide sequence 5′-UAGUAAUGAUAGGUAUCUGGAAU-3′ of SEQ ID NO:323.
23 . The dsRNA agent, or a salt thereof, 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 5, 6, 7, 15, and 17 on the sense strand, and positions 15 and 17 on the antisense strand, counting from the 5′-end of each strand.