SERPINA1 iRNA compositions and methods of use thereof
The invention relates to RNAi agents, e.g., double-stranded RNAi agents, targeting the Serpina1 gene, and methods of using such RNAi agents to inhibit expression of Serpina1 and methods of treating subjects having a Serpina1 associated disease, such as a liver disorder.
1. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of serine peptidase inhibitor, clade A member 1 (Serpina1) in a cell,
wherein said dsRNA agent comprises a sense strand and an antisense strand forming a double-stranded region,
wherein each strand is independently 19-30 nucleotides in length,
wherein said antisense strand comprises at least 19 contiguous nucleotides differing by no more than 3 nucleotides from any one of the antisense strand nucleotide sequences selected from the group consisting of
5′-UGGUAUUUUGUUCAAUCAUUAAG-3′ (SEQ ID NO:131);
5′-UUUUGUUCAAUCAUUAAGAAGAC-3′ (SEQ ID NO:133);
5′-AGACUUGGUAUUUUGUUCAAUCA-3′ (SEQ ID NO:135);
5′-AUUUUGUUCAAUCAUUAAGAAGA-3′ (SEQ ID NO: 137);
5′-AUUAAGAAGACAAAGGGUUUGUU-3′ (SEQ ID NO: 139);
5′-UGUUCAAUCAUUAAGAAGACAAA-3′ (SEQ ID NO: 141);
5′-AUCAUUAAGAAGACAAAGGGUUU-3′ (SEQ ID NO: 148);
5′-GAGACUUGGUAUUUUGUUCAAUC-3′ (SEQ ID NO: 150);
5′-AAGACAAAGGGUUUGUUGAACUU-3′ (SEQ ID NO: 152);
5′-AAGAGGGGAGACUUGGUAUUUUG-3′ (SEQ ID NO: 154);
5′-AGACAAAGGGUUUGUUGAACUUG-3′ (SEQ ID NO: 157);
5′-UUGGUAUUUUGUUCAAUCAUUAA-3′ (SEQ ID NO: 160);
5′-GACUUGGUAUUUUGUUCAAUCAU-3′ (SEQ ID NO: 179); and
5′-UGGGAUUCACCACUUUUCCCAUG-3′ (SEQ ID NO: 195),
wherein all of the nucleotides of said sense strand and all of the nucleotides of said antisense strand comprise a nucleotide modification, and
wherein at least one strand is conjugated to a ligand which is one or more GalNAc derivatives.
2. The dsRNA agent of claim 1 , wherein one of the 3 nucleotide differences in the nucleotide sequence of the antisense strand is a nucleotide mismatch in the seed region of the antisense strand.
3. The dsRNA agent of claim 1 , wherein the ligand is conjugated to the dsRNA agent through a bivalent or trivalent linker.
4. The dsRNA agent of claim 1 , wherein said agent further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage.
5. An isolated cell containing the dsRNA agent of claim 1 .
6. A pharmaceutical composition comprising the dsRNA agent of claim 1 .
7. A method of inhibiting Serpina1 expression 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 mRNA transcript of a Serpina1 gene, thereby inhibiting expression of the Serpina1 gene in the cell.
8. The dsRNA agent of claim 1 , wherein at least one of said nucleotide modifications is selected from the group consisting of 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 abasic nucleotide modification, a 2′-amino-nucleotide modification, a 2′-alkyl-nucleotide modification, a morpholino nucleotide modification, a phosphoramidate nucleotide modification, a non-natural base comprising nucleotide modification, a nucleotide comprising a 5′-phosphorothioate group nucleotide modification, and a terminal nucleotide linked to a cholesteryl derivative or a dodecanoic acid bisdecylamide group nucleotide modification.
9. The dsRNA agent of claim 1 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide; or a 3′ overhang of at least 2 nucleotides.
10. The dsRNA agent of claim 1 , wherein the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length.
11. The dsRNA agent of claim 3 , wherein the ligand is
12. The dsRNA agent of claim 11 , wherein the at least one ligand is conjugated to the 3′-end of the sense strand.
13. The dsRNA agent of claim 12 , wherein the dsRNA agent is conjugated to the ligand as shown in the following schematic
and, wherein X is O or S.
14. The dsRNA agent of claim 13 , wherein the X is O.
15. The dsRNA agent of claim 1 , wherein the sense strand comprises at least 19 contiguous nucleotides differing by no more than 3 nucleotides from any one of the sense strand nucleotide sequences selected from the group consisting of
5′-UAAUGAUUGAACAAAAUACCA-3′ (SEQ ID NO:43);
5′-CUUCUUAAUGAUUGAACAAAA-3′ (SEQ ID NO:45);
5′-AUUGAACAAAAUACCAAGUCU-3′ (SEQ ID NO:47);
5′-UUCUUAAUGAUUGAACAAAAU-3′ (SEQ ID NO:49);
5′-CAAACCCUUUGUCUUCUUAAU-3′ (SEQ ID NO:51);
5′-UGUCUUCUUAAUGAUUGAACA-3′ (SEQ ID NO:53);
5′-ACCCUUUGUCUUCUUAAUGAU-3′ (SEQ ID NO:60);
5′-UUGAACAAAAUACCAAGUCUC-3′ (SEQ ID NO:62);
5′-GUUCAACAAACCCUUUGUCUU-3′ (SEQ ID NO:64);
5′-AAAUACCAAGUCUCCCCUCUU-3′ (SEQ ID NO:66);
5′-AGUUCAACAAACCCUUUGUCU-3′ (SEQ ID NO:69);
5′-AAUGAUUGAACAAAAUACCAA-3′ (SEQ ID NO:72);
5′-GAUUGAACAAAAUACCAAGUC-3′ (SEQ ID NO: 91); and
5′-UGGGAAAAGUGGUGAAUCCCA-3′ (SEQ ID NO:107).
16. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of a serine peptidase inhibitor, clade A, member 1 (Serpina1) in a cell,
wherein the dsRNA agent comprises a sense strand and an antisense strand forming a double-stranded region,
wherein each strand is independently 19-30 nucleotides in length,
wherein the antisense strand comprises at least 19 contiguous nucleotides differing by no more than 3 nucleotides from any one of the antisense strand nucleotide sequences selected from the group consisting of
5′-UGGUAUUUUGUUCAAUCAUUAAG-3′ (SEQ ID NO:131);
5′-UUUUGUUCAAUCAUUAAGAAGAC-3′ (SEQ ID NO:133);
5′-AGACUUGGUAUUUUGUUCAAUCA-3′ (SEQ ID NO:135);
5′-AUUUUGUUCAAUCAUUAAGAAGA-3′ (SEQ ID NO: 137);
5′-AUUAAGAAGACAAAGGGUUUGUU-3′ (SEQ ID NO: 139);
5′-UGUUCAAUCAUUAAGAAGACAAA-3′ (SEQ ID NO: 141);
5′-AUCAUUAAGAAGACAAAGGGUUU-3′ (SEQ ID NO: 148);
5′-GAGACUUGGUAUUUUGUUCAAUC-3′ (SEQ ID NO: 150);
5′-AAGACAAAGGGUUUGUUGAACUU-3′ (SEQ ID NO: 152);
5′-AAGAGGGGAGACUUGGUAUUUUG-3′ (SEQ ID NO: 154);
5′-AGACAAAGGGUUUGUUGAACUUG-3′ (SEQ ID NO: 157);
5′-UUGGUAUUUUGUUCAAUCAUUAA-3′ (SEQ ID NO: 160);
5′-GACUUGGUAUUUUGUUCAAUCAU-3′ (SEQ ID NO: 179); and
5′-UGGGAUUCACCACUUUUCCCAUG-3′ (SEQ ID NO: 195),
wherein all of the nucleotides of said sense strand comprise a nucleotide modification,
wherein the sense strand comprises two phosphorothioate internucleotide linkages at the 5′-terminus,
wherein all of the nucleotides of said antisense strand comprise a nucleotide modification,
wherein the antisense strand comprises two phosphorothioate internucleotide linkages at the 5′-terminus and two phosphorothioate internucleotide linkages at the 3′-terminus, and
wherein at least one strand is conjugated to a GalNac ligand.
17. The dsRNA agent of claim 1 , wherein each strand is independently 19-23 nucleotides in length.
18. The dsRNA agent of claim 1 , wherein each strand is independently 19-25 nucleotides in length.