Apolipoprotein C3 (APOC3) iRNA compositions and methods of use thereof
The present invention relates to RNAi agents, e.g., double stranded RNA (dsRNA) agents, targeting the apolipoprotein C3 gene (APOC3). The invention also relates to methods of using such RNAi agents to inhibit expression of an APOC3 gene and to methods of preventing and treating an APOC3-associated disorder, e.g., hypertriglyceridemia, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, polycystic ovary syndrome, kidney disease, obesity, type 2 diabetes mellitus (insulin resistance), hypertension, artherosclerosis and pancreatitis.
1. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of apolipoprotein C3 in a cell,
wherein the dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of 5′-CUUAAAAGGGACAGUAUUCUA-3′ (SEQ ID NO:13), and the antisense strand comprises at least 15 contiguous nucleotides from the nucleotide sequence of 5′-UAGAAUACUGUCCCUUUUAAGCC-3′ (SEQ ID NO:14),
wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a modification selected from the group consisting of a 2′-O-methyl modification, a 2′-fluoro modification, and a deoxy-modification,
wherein the sense strand comprises 4 2′-fluoro modified nucleotides at nucleotides 7 and 9-11, counting from the 5′-end, and the antisense strand comprises 2 2′-fluoro modified nucleotides at nucleotides 14 and 16, counting from the 5′-end, and 3 2′-deoxy-modified nucleotides at nucleotides 2, 5, and 7, counting from the 5′-end,
wherein both the sense strand and the antisense strand independently further comprise at least one phosphorothioate or methylphosphonate internucleotide linkage, and
wherein at least one strand is conjugated to a ligand.
2. The dsRNA agent of claim 1 , wherein the sense strand comprises two phosphorothioate or methylphosphonate internucleotide linkages at the 5′-terminus.
3. The dsRNA agent of claim 1 , wherein the antisense strand comprises two phosphorothioate or methylphosphonate internucleotide linkages at both the 5′- and the 3′-terminus.
4. The dsRNA agent of claim 1 , wherein the sense strand comprises two phosphorothioate or methylphosphonate internucleotide linkages at the 5′-terminus and the antisense strand comprises two phosphorothioate or methylphosphonate internucleotide linkages at both the 5′- and the 3′-terminus.
5. The dsRNA agent of claim 1 , wherein the ligand is conjugated to the 3′-end of the sense strand.
6. The dsRNA agent of claim 1 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.
7. The dsRNA agent of claim 6 , wherein the ligand is one or more GalNAc derivatives attached through a monovalent, bivalent, or trivalent branched linker.
8. The dsRNA agent of claim 7 , wherein the ligand is
9. The dsRNA agent of claim 8 , wherein the dsRNA agent is conjugated to the ligand as shown in the following schematic
and, wherein X is O.
10. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of apolipoprotein C3 in a cell,
wherein the dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region, wherein the sense strand differs by no more than 3 modified nucleotides from the nucleotide sequence of 5′-csusuaaaAfgGfGfAfcaguauucua-3′ (SEQ ID NO: 15) and
wherein the antisense strand differs by no more than 3 modified nucleotides from the nucleotide sequence of 5′-usdAsgadAudAcuguccCfuUfuuaagscsc-3′ (SEQ ID NO:16),
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; dA is a 2′-deoxyadenosine-3′-phosphate nucleotide; and s is a phosphorothioate linkage.
11. The dsRNA agent of claim 10 , wherein the sense strand differs by no more than 2 modified nucleotides from the nucleotide sequence of 5′-csusuaaaAfgGfGfAfcaguauucua-3′ (SEQ ID NO: 15) and wherein the antisense strand differs by no more than 2 modified nucleotides from the nucleotide sequence of 5′-usdAsgadAudAcuguccCfuUfuuaagscsc-3′ (SEQ ID NO:16).
12. The dsRNA agent of claim 10 , wherein the sense strand differs by no more than 1 modified nucleotide from the nucleotide sequence of 5′-csusuaaaAfgGfGfAfcaguauucua-3′ (SEQ ID NO: 15) and wherein the antisense strand differs by no more than 1 modified nucleotides from the nucleotide sequence of 5′-usdAsgadAudAcuguccCfuUfuuaagscsc-3′ (SEQ ID NO:16).
13. The dsRNA agent of claim 10 , wherein the dsRNA agent is conjugated to the ligand as shown in the following schematic
and, wherein X is O.
14. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of apolipoprotein C3 in a cell,
wherein the dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand comprises the nucleotide sequence of 5′-csusuaaaAfgGfGfAfcaguauucua-3′ (SEQ ID NO: 15) and the antisense strand comprises the nucleotide sequence of 5′-usdAsgadAudAcuguccCfuUfuuaagscsc-3′ (SEQ ID NO:16),
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; dA is a 2′-deoxyadenosine-3′-phosphate nucleotide; and s is a phosphorothioate linkage.
15. The dsRNA agent of claim 14 , wherein the sense strand comprises the nucleotide sequence of 5′-csusuaaaAfgGfGfAfcaguauucuaL96-3′ (SEQ ID NO: 17) and the antisense strand comprises the nucleotide sequence of 5′-usdAsgadAudAcuguccCfuUfuuaagscsc-3′ (SEQ ID NO:16),
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; dA is a 2′-deoxyadenosine-3′-phosphate nucleotide; s is a phosphorothioate linkage, and L96 is N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol.
16. The dsRNA agent of claim 14 , wherein the sense strand comprises the nucleotide sequence of 5′-csusuaaaAfgGfGfAfcaguauucua-3′ (SEQ ID NO: 15) and the antisense strand comprises the nucleotide sequence of 5′-usdAsgadAudAcuguccCfuUfuuaagscsc-3′ (SEQ ID NO:16),
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; dA is a 2′-deoxyadenosine-3′-phosphate nucleotide; and s is a phosphorothioate linkage; and
wherein the 3′-end of the sense strand is conjugated to a ligand as shown in the following schematic:
wherein X is O.
17. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of apolipoprotein C3 in a cell,
wherein the dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand consists of the nucleotide sequence of 5′-csusuaaaAfgGfGfAfcaguauucua-3′ (SEQ ID NO: 15) and the antisense strand consists of the nucleotide sequence of 5′-usdAsgadAudAcuguccCfuUfuuaagscsc-3′ (SEQ ID NO:16),
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; dA is a 2′-deoxyadenosine-3′-phosphate nucleotide; and s is a phosphorothioate linkage; and
wherein the 3′-end of the sense strand is conjugated to a ligand as shown in the following schematic:
wherein X is O.
18. A pharmaceutical composition for inhibiting expression of a gene encoding apolipoprotein C3 (APOC3) comprising the dsRNA agent of claim 1 .
19. The pharmaceutical composition of claim 18 , wherein dsRNA agent is in an unbuffered solution.
20. The pharmaceutical composition of claim 19 , wherein the unbuffered solution is saline or water.
21. The pharmaceutical composition of claim 18 , wherein said dsRNA agent is in a buffer solution.
22. The pharmaceutical composition of claim 21 , wherein the buffer solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof.
23. The pharmaceutical composition of claim 22 , wherein the buffer solution is phosphate buffered saline (PBS).