Serapina1 iRNA compositions and methods of use thereof
View Patent ↗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 a 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, said antisense strand comprising a region of complementarity to an mRNA encoding Serpina1, wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of 5′-AAGAUAUUGGUGCUGUUGGACUG-3′ (SEQ ID NO:129),
wherein the sense strand and the antisense strand are each independently 19-25 nucleotides in length,
wherein substantially all of the nucleotides of said sense strand and substantially all of the nucleotides of said antisense strand are modified nucleotides, and
wherein said sense strand is conjugated to a ligand attached at the 3′-terminus.
2. The dsRNA agent of claim 1 , wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker.
3. The dsRNA agent of claim 1 , wherein said agent further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage.
4. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of a serine peptidase inhibitor, clade A, member 1 (Serpina1) in a cell, wherein said double stranded RNAi agent comprises a sense strand and an antisense strand forming a double-stranded region,
said antisense strand comprising a region of complementarity to an mRNA encoding Serpina1, wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of 5′-AAGAUAUUGGUGCUGUUGGACUG-3′ (SEQ ID NO:129),
wherein the sense strand and the antisense strand are each independently 19-25 nucleotides in length,
wherein substantially all of the nucleotides of said sense strand comprise a modification selected from the group consisting of a 2′-O-methyl modification and a 2′-fluoro modification,
wherein said sense strand comprises two phosphorothioate internucleotide linkages at the 5′-terminus,
wherein substantially all of the nucleotides of said antisense strand comprise a modification selected from the group consisting of a 2′-O-methyl modification and a 2′-fluoro modification,
wherein said antisense strand comprises two phosphorothioate internucleotide linkages at the 5′-terminus and two phosphorothioate internucleotide linkages at the 3′-terminus, and
wherein said sense strand is conjugated to one or more GalNAc derivatives attached through a branched bivalent or trivalent linker at the 3′-terminus.
5. A pharmaceutical composition comprising the dsRNA agent of claim 1 or 4 .
6. A method of inhibiting serine peptidase inhibitor, clade A, member 1 (Serpina1) expression in a cell, the method comprising:
(a) contacting the cell with the dsRNA agent of claim 1 or 4 or the pharmaceutical composition of claim 5 ; 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.
7. A method of treating a subject having a serine peptidase inhibitor, clade A, member 1 (Serpina1) deficiency variant-associated disease, comprising administering to the subject a therapeutically effective amount of the dsRNA agent of claim 1 or 4 or the pharmaceutical composition of claim 5 , thereby treating said subject.
8. The method of claim 7 , wherein the Serpina1 associated disease is a liver disorder.
9. A method of treating a subject having a serine peptidase inhibitor, clade A, member 1 (Serpina1) deficiency variant-associated liver disorder to reduce the progression of the liver disorder to hepatocellular carcinoma, comprising administering to the subject a therapeutically effective amount of the dsRNA agent of claim 1 or 4 or the pharmaceutical composition of claim 5 , thereby treating the subject the subject.
10. A method of reducing the accumulation of misfolded serine peptidase inhibitor, clade A, member 1 (Serpina1) in the liver of a subject having a Serpina1 deficiency variant, comprising administering to the subject a therapeutically effective amount of the dsRNA agent of claim 1 or 4 or the pharmaceutical composition of claim 5 , thereby reducing the accumulation of misfolded Serpina1 in the liver of the subject.
11. The dsRNA of claim 1 , wherein all of the nucleotides of said sense strand and all of the nucleotides of said antisense strand are modified nucleotides.
12. The dsRNA agent of claim 1 , wherein at least one of said modified nucleotides is selected from the group consisting of 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 abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, a non-natural base comprising nucleotide, a nucleotide comprising a 5′-phosphorothioate group, and a terminal nucleotide linked to a cholesteryl derivative or a dodecanoic acid bisdecylamide group.
13. The dsRNA agent of claim 12 , wherein the modifications on the nucleotides are 2′-O-methyl or 2′-fluoro modifications.
14. The dsRNA agent of any claim 1 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide.
15. The dsRNA agent of claim 1 , wherein at least one strand comprises a 3′ overhang of at least 2 nucleotides.
16. The dsRNA agent of claim 3 , wherein said RNAi agent comprises 6-8 phosphorothioate internucleotide linkages.
17. The dsRNA of claim 16 , wherein the antisense strand comprises two phosphorothioate internucleotide linkages at the 5′-terminus and two phosphorothioate internucleotide linkages at the 3′-terminus, and the sense strand comprises at least two phosphorothioate internucleotide linkages at either the 5′-terminus or the 3′-terminus.
18. The dsRNA agent of claim 2 , wherein the ligand is
19. The dsRNA agent of claim 18 , wherein the agent is conjugated to the ligand as shown in the following schematic
and, wherein X is O or S.
20. The dsRNA agent of claim 19 , wherein the X is O.
21. The dsRNA agent of claim 1 , wherein the double stranded region is 17-25 nucleotide pairs in length.
22. The dsRNA agent of claim 1 , wherein the region of complementarity consists of the nucleotide sequence of 5′-AAGAUAUUGGUGCUGUUGGACUG-3′ (SEQ ID NO:129).
23. The dsRNA agent of claim 1 , wherein the agent comprises a sense strand comprising the nucleotide sequence of 5′-GUCCAACAGCACCAAUAUCUU-3′ (SEQ ID NO:41), and an antisense strand comprising the nucleotide sequence of 5′-AAGAUAUUGGUGCUGUUGGACUG-3′ (SEQ ID NO:129).
24. The dsRNA agent of claim 23 , wherein the sense strand comprises 5′-GfsusCfcAfaCfaGfCfAfcCfaAfuAfuCfuUf-3′ (SEQ ID NO:217) and the antisense strand comprises 5′-asAfsgAfuAfuUfgGfugcUfgUfuGfgAfcsUfsg-3′ (SEQ ID NO:305),
wherein a, g, c, and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; Af, Gf, Cf and Uf are 2′-fluoro A, G, C and U, respectively; and s is a phosphorothioate linkage.
25. A cell containing the dsRNA agent of claim 1 or 4 .
26. The dsRNA agent of claim 4 , wherein all of the nucleotides of said sense strand and all of the nucleotides of said antisense strand comprise a modification.
27. The method of claim 6 , wherein cell is within a subject.
28. The method of claim 27 , wherein subject is human.
29. The method of claim 8 , wherein the liver disorder is selected from the group consisting of chronic liver disease, liver inflammation, cirrhosis, liver fibrosis, and hepatocellular carcinoma.
30. The dsRNA agent of claim 1 or claim 4 , wherein the sense strand and the antisense strand are separate nucleic acid molecules.
31. The dsRNA agent of claim 1 or claim 4 , wherein the sense strand and the antisense strand are portions of a single nucleic acid molecule.
32. The dsRNA agent of claim 31 , wherein the sense strand and the antisense strand are connected by an uninterrupted chain of nucleotides between the 3′-end of the sense strand and the 5′-end of the antisense strand.
33. The dsRNA agent of claim 31 , wherein the sense strand and the antisense strand are connected by an uninterrupted chain of nucleotides between the 3′-end of the antisense strand and the 5′-end of the sense strand.
34. The dsRNA agent of claim 31 , wherein the sense strand and the antisense strand are covalently connected by a linker between the 3′-end of the sense strand and the 5′-end of the antisense strand.
35. The dsRNA agent of claim 31 , wherein the sense strand and the antisense strand are covalently connected by a linker between the 3′-end of the antisense strand and the 5′-end of the sense strand.
36. The method of claim 7 or 9 , wherein the subject has one or more of a Serpina1 Z allele.