NZ736316B2 - Angiopoietin-like 3 (angptl3) irna compositions and methods of use thereof - Google Patents

Abstract

The invention relates to double-stranded ribonucleic acid (dsRNA) compositions targeting the ANGPTL3 gene, as well as methods of inhibiting epression of ANGPTL3 and methods of treating subjects having a disorder of lipid metabolism, such as hyperlipidemia or hypertriglyceridemia, using such dsRNA compositions.

Claims (36)

We claim:

1. A double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of angiopoietin-like 3 (ANGPTL3), or a salt thereof, wherein the dsRNA agent, or a salt thereof, comprises a sense strand and an antisense 5 strand g a double ed region, wherein the antisense strand comprises the nucleotide sequence 5’- UGAAGUUACUUCUGGGU-3’ (SEQ ID NO:511) or 5’- UUUCUACAAAAGUUUUAAGUGAA (SEQ ID NO:529), n all of the nucleotides of the sense strand and all of the nucleotides of the 10 antisense strand comprise a nucleotide modification, n at least one of the nucleotide modifications is selected from the group consisting of a ethyl nucleotide modification, a 2’-fluoro nucleotide modification, a nucleotide comprising a 5'-phosphorothioate group modification, and a 2’-amino nucleotide modification, and 15 wherein a ligand comprising an N-acetylgalactosamine (GalNAc) derivative is conjugated to at least one strand of the dsRNA agent.

2. The dsRNA agent, or a salt thereof, of claim 1, wherein the sense strand is 19- 25 nucleotides in length. 20

3. The RNAi agent, or a salt f, of claim 1 or 2, wherein the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length.

4. The dsRNA agent, or a salt thereof, of any one of claims 1-3, wherein at least 25 one strand comprises a 3’ overhang of at least 1 nucleotide.

5. The dsRNA agent, or a salt thereof, of any one of claims 1-4, wherein at least one strand comprises a 3’ overhang of at least 2 nucleotides.

6. The dsRNA agent, or a salt thereof, of any one of claims 1-5, n at least one of the 5’-end or the 3’-end of the sense strand of the dsRNA agent, or a salt thereof, is a blunt end. 21328918_1 (GHMatters) P107021.NZ

7. The dsRNA agent, or a salt thereof, of any one of claims 1-6, wherein the sense and nse strands comprise nucleotide sequences selected from the group consisting 5 (i) 5’-CCAGAAGUAACUUCACUUAAA-3’ (SEQ ID NO:470) and 5’- UUUAAGUGAAGUUACUUCUGGGU-3’ (SEQ ID NO:511); and (ii) 5’-CACUUAAAACUUUUGUAGAAA-3’ (SEQ ID NO:488) and 5’- UUUCUACAAAAGUUUUAAGUGAA-3’ (SEQ ID NO:529).

8. A double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of 10 Angiopoietin-like 3 (ANGPTL3), or a salt thereof, wherein the dsRNA agent, or a salt thereof, ses a sense strand and an antisense strand forming a double stranded region, wherein the sense strand ses the nucleotide ce 5’- CCAGAAGUAACUUCACUUAAA-3’ (SEQ ID NO:470) and the antisense strand 15 comprises the nucleotide sequence 5’-UUUAAGUGAAGUUACUUCUGGGU-3’ (SEQ ID wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a nucleotide modification, wherein at least one of the nucleotide modifications is selected from the group 20 consisting of a 2'-O-methyl nucleotide modification, a 2’-fluoro nucleotide modification, a nucleotide comprising a 5'-phosphorothioate group modification, and a no nucleotide modification, and wherein a ligand sing an N-acetylgalactosamine (GalNAc) derivative is ated to at least one strand of the dsRNA agent, or a salt thereof. 25

9. A double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of Angiopoietin-like 3 (ANGPTL3), or a salt thereof, wherein the dsRNA agent, or a salt thereof, comprises a sense strand and an antisense strand, 30 wherein the sense strand comprises the nucleotide sequence 5’- CACUUAAAACUUUUGUAGAAA-3’ (SEQ ID ) and the antisense strand 21328918_1 (GHMatters) P107021.NZ ses the nucleotide sequence CUACAAAAGUUUUAAGUGAA (SEQ ID NO:529), wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a nucleotide modification, wherein at least one of the nucleotide modifications is ed from the group 5 consisting of a 2'-O-methyl nucleotide modification, a 2’-fluoro nucleotide modification, a nucleotide comprising a 5'-phosphorothioate group modification, and a 2’-amino nucleotide modification, and wherein a ligand comprising an N-acetylgalactosamine (GalNAc) derivative is conjugated to at least one strand of the dsRNA agent, or a salt thereof. 10

10. The dsRNA agent, or a salt thereof, of any one of claims 1-9, wherein all of the nucleotides comprise a modification selected from the group consisting of a ethyl cation and a ro modification.

11. The dsRNA agent, or a salt thereof, of any one of claims 1-10, further comprising at least one phosphorothioate internucleotide linkage.

12. The dsRNA agent, or a salt thereof, of claim 11, wherein the orothioate or methylphosphonate internucleotide linkage is at the minus of one strand. 20

13. The dsRNA agent, or a salt thereof, of claim 12, wherein the strand is the antisense strand.

14. The dsRNA agent of claim 11, wherein the dsRNA agent, or a salt thereof, 25 comprises 6-8 phosphorothioate internucleotide linkages.

15. The dsRNA agent, or a salt thereof, of any one of claims 1-14, wherein the ligand is conjugated to the 3’ end of the sense strand of the dsRNA, or a salt thereof.

16. The dsRNA agent, or a salt thereof, of any one of claims 1-15, wherein the ligand comprising the GalNAc derivative is attached to the dsRNA, or a salt thereof, agent through a linker. 21328918_1 (GHMatters) P107021.NZ

17. The dsRNA agent, or a salt thereof, of claim 16, wherein the ligand comprising the GalNAc derivative is ed to the dsRNA agent, or a salt thereof, through a bivalent or trivalent linker.

18. The dsRNA agent, or a salt thereof, of claim 17, wherein the ligand is HO OH O H H HO O N N O HO OH O H H HO O N N O O O O HO OH HO O N N O AcHN H H O .

19. The dsRNA agent, or a salt thereof, of claim 18, wherein the dsRNA agent, or a salt thereof, is conjugated to the ligand as shown in the following tic 10 , wherein X is O or S.

20. An isolated cell containing the dsRNA agent, or a salt thereof, of any one of claims 1-19.

21. A pharmaceutical composition for ting expression of an ANGPTL3 gene comprising the dsRNA agent, or a salt thereof, of any one of claims 1-19. 21328918_1 (GHMatters) P107021.NZ

22. An in vitro method of inhibiting 3 expression in a cell, the method sing: (a) ting the cell with the dsRNA agent, or a salt thereof, of any one of claims 1-19 or the pharmaceutical composition of claim 21; and (b) maintaining the cell produced in step (a) for a time sufficient to obtain 5 degradation of the mRNA transcript of an ANGPTL3 gene, thereby inhibiting expression of the ANGPTL3 gene in the cell.

23. Use of the dsRNA agent, or a salt thereof, of any one of claims 1-19 or the pharmaceutical ition of claim 21, in the manufacture of a medicament for treating a 10 human subject having a disorder that would t from a reduction in ANGPTL3 expression.

24. The use of claim 23, wherein the disorder is a disorder of lipid metabolism.

25. The use of claim 24, wherein the disorder of lipid metabolism is selected from the group consisting of hypertriglyceridemia, y, hyperlipidemia, atherosclerosis, diabetes, cardiovascular disease, or coronary artery disease.

26. The use of claim 24, wherein the disorder of lipid metabolism is 20 hyperlipidemia or hypertriglyceridemia.

27. The use of any one of claims 23-26, wherein the use further comprises use of an additional therapeutic.

28. The use of claim 27, wherein the additional therapeutic is a statin.

29. The use of any one of claims 23-28, wherein the dsRNA agent, or a salt thereof, or pharmaceutical composition is for subcutaneous administration.

30. The use of any one of claims 23-29, wherein the dsRNA agent, or a salt thereof, or pharmaceutical composition is for administration at a dose of about 0.01 mg/kg to about 50 mg/kg. 21328918_1 (GHMatters) P107021.NZ

31. The use of any one of claims 23-30, wherein administration. of the dsRNA agent, or a salt thereof, or pharmaceutical composition causes a se in one or more serum lipid and/or a decrease in ANGPTL3 protein accumulation.

32. Use of the dsRNA agent, or a salt thereof, of any one of claims 1-19 or the pharmaceutical composition of claim 21, in the manufacture of a medicament for ting the expression of ANGPTL3 in a human subject.

33. The use of claim 32, wherein the subject has a disorder of lipid metabolism. 10

34. The use of any one of claims 32-33, wherein the use further comprises use of an additional therapeutic.

35. The use of claim 34, wherein the additional the rapeutic is a statin. 15

36. The use of any one of claims 32-35, wherein the dsRNA agent, or a salt thereof, or pharmaceutical composition is for subcutaneous stration. 21328918_1 ters) P107021.NZ ~\\\\\\\\\\\\\\\\m Optimized ~\\\10 ANG—GaINAc ....................... “““““““