Angiopoietin-like 3 (ANGPTL3) iRNA compositions and methods of use thereof
The invention relates to double-stranded ribonucleic acid (dsRNA) compositions targeting the ANGPTL3 gene, as well as methods of inhibiting expression of ANGPTL3 and methods of treating subjects having a disorder of lipid metabolism, such as hyperlipidemia or hypertriglyceridemia, using such dsRNA compositions.
1. A double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of angiopoietin-like 3 (ANGPTL3), wherein the dsRNA comprises a sense strand and an antisense strand forming a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence 5′-UUGAGUUGAGUUCAAGUGACAUA 3′ (SEQ ID NO: 308), wherein substantially all of the nucleotides of the sense strand and substantially all of the nucleotides of the antisense strand comprise a modification selected from the group consisting of a 2′-O-methyl modification and a 2′-fluoro modification, and wherein the sense strand comprises no more that four 2′-fluoro modified nucleotides, and the antisense strand comprises no more than six 2′-fluoro modified nucleotides.
2. The dsRNA agent of claim 1 , wherein all of the nucleotides of the sense strand are modified nucleotides; or wherein all of the nucleotides of the antisense strand are modified nucleotides.
3. The dsRNA agent of claim 1 , further comprising at least one phosphorothioate internucleotide linkage.
4. The dsRNA agent of claim 1 , wherein each strand is no more than 30 nucleotides in length.
5. The dsRNA agent of claim 1 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide.
6. The dsRNA agent of claim 1 , further comprising a ligand.
7. The dsRNA agent of claim 6 , wherein the ligand is conjugated to the 3′ end of the sense strand of the dsRNA agent.
8. The dsRNA agent of claim 7 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.
9. The dsRNA agent of claim 8 , wherein the ligand is
10. The dsRNA agent of claim 1 , wherein the double-stranded region is 15-30 nucleotide pairs in length.
11. The dsRNA agent of claim 1 , wherein each strand is 15-30 nucleotides in length.
12. The dsRNA agent of claim 1 , wherein each of the sense strand and the antisense strand independently are 21 to 23 nucleotides in length.
13. The dsRNA agent of claim 1 , wherein the sense strand has a total of 21 nucleotides and the antisense strand has a total of 23 nucleotides.
14. The dsRNA agent of claim 1 , wherein the antisense strand comprises the nucleotide sequence 5′-UUGAGUUGAGUUCAAGUGACAUA 3′ (SEQ ID NO: 308).
15. The dsRNA agent of claim 1 , wherein the antisense strand consists of the nucleotide sequence 5′-UUGAGUUGAGUUCAAGUGACAUA-3′ (SEQ ID NO: 308).
16. The dsRNA agent of claim 1 , wherein the sense strand comprises the nucleotide sequence 5′-UGUCACUUGAACUCAACUCAA-3′ (SEQ ID NO:296) and the antisense strand comprises the nucleotide sequence 5′-UUGAGUUGAGUUCAAGUGACAUA-3′ (SEQ ID NO:308).
17. The dsRNA agent of claim 16 , wherein the sense strand comprises the nucleotide sequence 5′-usgsucacUfuGfAfAfcucaacucaa-3′ (SEQ ID NO:272) and the antisense strand comprises the nucleotide sequence 5′-usUfsgagUfuGfAfguucAfaGfugacasusa-3′ (SEQ ID NO:284),
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C and U; and s is a phosphorothioate linkage.
18. The dsRNA agent of claim 17 , further comprising a ligand.
19. The dsRNA of claim 18 , wherein the ligand is one or more GalNAc derivatives attached through a monovalent, a bivalent, or a trivalent branched linker.
20. The dsRNA agent of claim 19 , wherein the ligand is
21. The dsRNA agent of claim 18 , wherein the ligand is attached to the 3′ end of the sense strand.
22. The dsRNA agent of claim 1 , wherein at least one strand comprises a 3′ overhang of at least 2 nucleotides.
23. The dsRNA agent of claim 1 , wherein the double-stranded region is 17-23 nucleotide pairs in length.
24. The dsRNA agent of claim 1 , wherein the double-stranded region is 17-25 nucleotide pairs in length.
25. The dsRNA agent of claim 1 , wherein the double-stranded region is 23-27 nucleotide pairs in length.
26. The dsRNA agent of claim 1 , wherein the double-stranded region is 19-21 nucleotide pairs in length.
27. The dsRNA agent of claim 1 , wherein the double-stranded region is 21-23 nucleotide pairs in length.
28. The dsRNA agent of claim 1 , wherein each strand is 19-30 nucleotides in length.
29. The dsRNA agent of claim 9 or 20 , wherein the dsRNA agent is conjugated to the ligand as shown in the following schematic
wherein X is O or S.
30. The dsRNA agent of claim 29 , wherein the X is O.
31. The dsRNA agent of claim 1 , wherein the antisense strand comprises the nucleotide sequence 5′-UTGAGUUGAGUTCAAGUGACAUA-3′ (SEQ ID NO: 387).
32. The dsRNA agent of claim 1 , wherein the antisense strand consists of the nucleotide sequence 5′-UTGAGUUGAGUTCAAGUGACAUA-3′ (SEQ ID NO: 387).
33. The dsRNA agent of claim 1 , wherein the sense strand comprises the nucleotide sequence 5′-UGUCACUUCAACUCAACUCAA-3′ (SEQ ID NO:366) and the antisense strand comprises the nucleotide sequence 5′-UTGAGUUGAGUTCAAGUGACAUA-3′ (SEQ ID NO:387).
34. The dsRNA agent of claim 33 , wherein the sense strand comprises the nucleotide sequence 5′-usgsucadCuudCaacucaa(Cgn)ucaa-3′ (SEQ ID NO:324) and the antisense strand comprises the nucleotide sequence 5′-usdTsgaguugagudTcdAagugacasusa-3′ (SEQ ID NO:345), wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C and U; dT is deoxythymidine; dA is deoxyadenosine; (Cgn) is Cytidine-glycol nucleic acid (GNA); and s is a phosphorothioate linkage.
35. The dsRNA agent of claim 34 , further comprising a ligand.
36. A pharmaceutical composition for inhibiting expression of an ANGPTL3 gene comprising the dsRNA agent of claim 35 .
37. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of angiopoietin-like 3 (ANGPTL3), wherein the dsRNA comprises a sense strand and an antisense strand forming a double stranded region, wherein the sense strand consists of the nucleotide sequence 5′-usgsucacUfuGfAfAfcucaacucaaL96-3′ (SEQ ID NO:272) and the antisense strand consists of the nucleotide sequence 5′-usUfsgagUfuGfAfguucAfaGfugacasusa-3′ (SEQ ID NO:284),
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C and U; s is a phosphorothioate linkage; and L96 is N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol.
38. A pharmaceutical composition for inhibiting expression of an ANGPTL3 gene comprising the dsRNA agent of claim 37 .
39. An isolated cell containing the dsRNA agent of claim 1 or 17 .
40. A pharmaceutical composition for inhibiting expression of an ANGPTL3 gene comprising the dsRNA agent of claim 1 or 17 .
41. A method of inhibiting ANGPTL3 expression in a cell, the method comprising:
(a) contacting the cell with the dsRNA agent of any one of claims 1 , 17 , and 37 ; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of an ANGPTL3 gene, thereby inhibiting expression of the ANGPTL3 gene in the cell.
42. The method of claim 41 , wherein said cell is within a subject.
43. The method of claim 42 , wherein the subject is a human.
44. The method of claim 43 , wherein the human subject suffers from a disorder of lipid metabolism.
45. The method of claim 44 , wherein the disorder of lipid metabolism is hyperlipidemia or hypertriglyceridemia.
46. A method of treating a subject having a disorder that would benefit from a reduction in ANGPTL3 expression, comprising administering to the subject a therapeutically effective amount of the dsRNA agent of any one of claims 1 , 17 , and 37 , thereby treating said subject.