Angiopoietin-like 3 (ANGPTL3) iRNA compositions and methods of use thereof
The present invention relates to RNAi agents, e.g., double stranded RNA (dsRNA) agents, targeting the Angiopoietin-like 3 (ANGPTL3) gene. The invention also relates to methods of using such RNAi agents to inhibit expression of an ANGPTL3 gene and to methods of preventing and treating an ANGPTL3-associated disorder, e.g., a disorder of lipid metabolism, such as hyperlipidemia or hypertriglyceridemia.
1. A double stranded ribonucleic acid (dsRNA) agent, or a salt thereof, for inhibiting expression of Angiopoietin-like 3 (ANGPTL3) in a cell,
wherein the dsRNA agent, or salt thereof, comprises a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand comprises at least 19 contiguous nucleotides of the nucleotide sequence 5′-AAGCUCCUUCUUUUUAUUGUU-3′ (SEQ ID NO: 46) and the antisense strand comprises at least 21 contiguous nucleotides of the nucleotide sequence of 5′-AACAAUAAAAAGAAGGAGCUUGG-3′ (SEQ ID NO: 661),
wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a modification, and
wherein at least one of the modifications is a 2′-deoxy-nucleotide modification.
2. The dsRNA agent, or salt thereof, of claim 1 , wherein the double stranded region is 19-30 nucleotide pairs in length.
3. The dsRNA agent, or salt thereof, of claim 2 , wherein the double stranded region is 21-23 nucleotide pairs in length.
4. The dsRNA agent, or salt thereof, of claim 1 wherein each strand is independently 19-30 nucleotides in length.
5. The dsRNA agent, or salt thereof, of claim 1 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide.
6. The dsRNA agent, or salt thereof, of claim 1 , further comprising a ligand.
7. The dsRNA agent, or salt thereof, of claim 6 , wherein the ligand is conjugated to the 3′ end of the sense strand of the dsRNA agent, or salt thereof.
8. The dsRNA agent, or salt thereof, of claim 6 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.
9. The dsRNA agent, or salt thereof, of claim 6 , wherein the ligand is one or more GalNAc derivatives attached through a monovalent, bivalent, or trivalent branched linker.
10. The dsRNA agent, or salt thereof, of claim 9 , wherein the ligand is
11. The dsRNA agent, or salt thereof, of claim 10 , wherein the dsRNA agent, or salt thereof, is conjugated to the ligand as shown in the following schematic
and, wherein X is O or S.
12. The dsRNA agent, or salt thereof, of claim 11 , wherein the X is O.
13. The dsRNA agent, or salt thereof, of claim 1 , wherein the agent further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage.
14. The dsRNA agent, or salt thereof, of claim 13 , wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the 3′-terminus of one strand; the 5′-terminus of one strand; or at both the 5′- and 3′-terminus of one strand.
15. The dsRNA agent, or salt thereof, of claim 1 , wherein the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length.
16. The dsRNA agent, or salt thereof, of claim 1 , wherein the sense strand comprises the nucleotide sequence 5′-AAGCUCCUUCUUUUUAUUGUU-3′ (SEQ ID NO: 46).
17. The dsRNA agent, or salt thereof, of claim 1 , wherein the sense strand comprises the nucleotide sequence 5′-AAGCUCCUUCUUUUUAUUGUU-3′ (SEQ ID NO: 46) and the antisense strand comprises the nucleotide sequence 5′-AACAAUAAAAAGAAGGAGCUUGG-3′ (SEQ ID NO: 661).
18. The dsRNA agent, or salt thereof, of claim 1 , wherein the sense strand consists of the nucleotide sequence 5′-AAGCUCCUUCUUUUUAUUGUU-3′ (SEQ ID NO: 46) and the antisense strand consists of the nucleotide sequence 5′-AACAAUAAAAAGAAGGAGCUUGG-3′ (SEQ ID NO: 661).
19. The dsRNA agent, or salt thereof, of claim 1 , wherein the sense strand comprises the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ (SEQ ID NO: 20) and the antisense strand comprises the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ (SEQ ID NO: 19),
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Cf and Uf are 2′-fluoro (2′-F) C and U, respectively; and s is a phosphorothioate linkage.
20. The dsRNA agent, or salt thereof, of claim 1 , wherein the sense strand consists of the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ (SEQ ID NO: 20) and the antisense strand consists of the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ (SEQ ID NO: 19),
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Cf and Uf are 2′-fluoro (2′-F) C and U, respectively; s is a phosphorothioate linkage; and L96 is N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol.
21. A double stranded ribonucleic acid (dsRNA) agent, or a salt thereof, for inhibiting expression of Angiopoietin-like 3 (ANGPTL3) in a cell, comprising a sense strand and an antisense strand forming a double stranded region,
wherein the nucleotide sequence of the sense strand differs by no more than 4 bases from the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ (SEQ ID NO: 20) and the nucleotide sequence of the antisense strand differs by no more than 4 bases from the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ (SEQ ID NO: 19),
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Cf and Uf are 2′-fluoro (2′-F) C and U, respectively; and s is a phosphorothioate linkage.
22. The dsRNA agent of claim 21 , wherein the nucleotide sequence of the sense strand differs by no more than 3 bases from the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ (SEQ ID NO: 20) and the nucleotide sequence of the antisense strand differs by no more than 3 bases from the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ (SEQ ID NO: 19),
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Cf and Uf are 2′-fluoro (2′-F) C and U, respectively; and s is a phosphorothioate linkage.
23. The dsRNA agent of claim 21 , wherein the nucleotide sequence of the sense strand differs by no more than 2 bases from the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ (SEQ ID NO: 20) and the nucleotide sequence of the antisense strand differs by no more than 2 bases from the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ (SEQ ID NO: 19),
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Cf and Uf are 2′-fluoro (2′-F) C and U, respectively; and s is a phosphorothioate linkage.
24. The dsRNA agent of claim 21 , wherein the nucleotide sequence of the sense strand differs by no more than 1 base from the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ (SEQ ID NO: 20) and the nucleotide sequence of the antisense strand differs by no more than 1 base from the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ (SEQ ID NO: 19),
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Cf and Uf are 2′-fluoro (2′-F) C and U, respectively; and s is a phosphorothioate linkage.
25. The dsRNA agent of claim 21 , wherein the ligand is conjugated to the 3′ end of the sense strand of the dsRNA agent.
26. A double stranded ribonucleic acid (dsRNA) agent, or a salt thereof, for inhibiting expression of Angiopoietin-like 3 (ANGPTL3) in a cell, comprising a sense strand and an antisense strand forming a double stranded region,
wherein the nucleotide sequence of the sense strand comprises the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ (SEQ ID NO: 20) and the nucleotide sequence of the antisense strand comprises the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ (SEQ ID NO: 19),
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Cf and Uf are 2′-fluoro (2′-F) C and U, respectively; and s is a phosphorothioate linkage.
27. The dsRNA agent, or salt thereof, of claim 26 , further comprising a ligand.
28. A double stranded ribonucleic acid (dsRNA) agent, or a salt thereof, for inhibiting expression of Angiopoietin-like 3 (ANGPTL3) in a cell, comprising a sense strand and an antisense strand forming a double stranded region,
wherein the nucleotide sequence of the sense strand comprises the nucleotide sequence 5′-asasgcuccuUfCfUfuuuuauuguu-3′ (SEQ ID NO: 20) and the nucleotide sequence of the antisense strand comprises the nucleotide sequence 5′-asdAscadAudAaaaadGaAfggagcuusgsg-3′ (SEQ ID NO: 19),
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; dA and dG, are 2′-deoxy A and G, respectively; Cf and Uf are 2′-fluoro (2′-F) C and U, respectively; s is a phosphorothioate linkage, and
wherein a ligand is conjugated to the 3′-end of the sense strand as shown in the following schematic
wherein X is O.
29. The dsRNA agent, or a salt thereof, of claim 28 , which is in a salt form.
30. A pharmaceutical composition for inhibiting expression of a gene encoding ANGPTL3 comprising the dsRNA agent, or salt thereof, of claim 28 .