Patatin-like phospholipase domain containing 3 (PNPLA3) iRNA compositions and methods of use thereof

Patent metadata

Field	Value
Publication number	US-11052103-B2
Application number	US-201916267615-A
Country	US
Kind code	B2
Filing date	Feb 5, 2019
Priority date	Feb 13, 2015
Publication date	Jul 6, 2021
Grant date	Jul 6, 2021

Abstract

The present invention relates to RNAi agents, e.g., double stranded RNAi agents, targeting the Patatin-Like Phospholipase Domain Containing 3 (PNPLA3) gene, and methods of using such RNAi agents to inhibit expression of a PNPLA3 gene and methods of treating subjects having Nonalcoholic Fatty Liver Disease (NAFLD) and/or a PNPLA3-associated disorder.

First claim

We claim:

1. A double stranded ribonucleic acid (RNAi) agent for inhibiting expression of Patatin-Like Phospholipase Domain Containing 3 (PNPLA3),
wherein said double stranded RNAi agent 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 complement of nucleotides 577-634 of SEQ ID NO:1.

2. The double stranded RNAi agent of claim 1 , wherein said double stranded RNAi agent comprises at least one modified nucleotide.

3. The double stranded RNAi agent of claim 2 , wherein the at least one modified nucleotide is selected from the group consisting of a deoxy-nucleotide, 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 unlocked nucleotide, a conformationally restricted nucleotide, a constrained ethyl nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-O-allyl-modified nucleotide, 2′-C-alkyl-modified nucleotide, 2′-hydroxyl-modified nucleotide, a 2′-methoxyethyl modified nucleotide, a 2′-O-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, a non-natural base comprising nucleotide, a tetrahydropyran modified nucleotide, a 1,5-anhydrohexitol modified nucleotide, a cyclohexenyl modified nucleotide, a nucleotide comprising a phosphorothioate group, a nucleotide comprising a methylphosphonate group, a nucleotide comprising a 5′-phosphate, and a nucleotide comprising a 5′-phosphate mimic.

4. The double stranded RNAi agent of claim 1 , wherein each strand is no more than 30 nucleotides in length.

5. The double stranded RNAi agent of claim 1 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide.

6. The double stranded RNAi agent of claim 1 , further comprising a ligand.

7. The double stranded RNAi agent of claim 6 , wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker.

8. The double stranded RNAi agent of claim 7 , wherein the ligand is

9. The double stranded RNAi agent of claim 1 , wherein said double stranded RNAi agent further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage.

10. A cell containing the double stranded RNAi agent of claim 1 .

11. A pharmaceutical composition for inhibiting expression of a PNPLA3 gene comprising the double stranded RNAi agent of claim 1 .

12. A method of inhibiting PNPLA3 expression in a cell, the method comprising:
(a) contacting the cell with the double stranded RNAi agent of claim 1 or a pharmaceutical composition of claim 11 ; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a PNPLA3 gene, thereby inhibiting expression of the PNPLA3 gene in the cell.

13. The method of claim 12 , wherein said cell is within a subject.

14. The method of claim 13 , wherein the subject is a human.

15. A method of treating a subject having a disease or disorder that would benefit from reduction in PNPLA3 expression, the method comprising administering to the subject a therapeutically effective amount of the double stranded RNAi agent of claim 1 or a pharmaceutical composition of claim 11 , thereby treating said subject.

16. A method of preventing at least one symptom in a subject having a disease or disorder that would benefit from reduction in PNPLA3 expression, the method comprising administering to the subject a prophylactically effective amount of the double stranded RNAi agent of claim 1 or a pharmaceutical composition of claim 11 , thereby preventing at least one symptom in the subject having a disorder that would benefit from reduction in PNPLA3 expression.

17. The method of claim 15 , wherein the disorder is a PNPLA3-associated disease.

18. The method of claim 15 , further comprising administering an anti-PNPLA3 antibody, or antigen-binding fragment thereof, to the subject.

19. A double stranded ribonucleic acid (RNAi) agent for inhibiting expression of Patatin-Like Phospholipase Domain Containing 3 (PNPLA3),
wherein the double stranded RNAi agent comprises a sense strand and an antisense strand forming a double stranded region,
wherein the antisense strand comprises at least 16 contiguous nucleotides differing by no more than 3 nucleotides from the complement of nucleotides 577-634 of SEQ ID NO:1.

20. A double stranded ribonucleic acid (RNAi) agent for inhibiting expression of Patatin-Like Phospholipase Domain Containing 3 (PNPLA3),
wherein the double stranded RNAi agent comprises a sense strand and an antisense strand forming a double stranded region,
wherein the antisense strand comprises at least 17 contiguous nucleotides differing by no more than 3 nucleotides from the complement of nucleotides 577-634 of SEQ ID NO:1.

21. The double stranded RNAi agent of claim 1 , wherein each strand is independently 17-25 nucleotides in length.

22. The double stranded RNAi agent of claim 1 , wherein each strand is independently 19-25 nucleotides in length.

23. The double stranded RNAi agent of claim 1 , wherein each strand is independently 19-23 nucleotides in length.

24. The double stranded RNAi agent of claim 1 , wherein each strand is independently 21 to 23 nucleotides in length.

25. The double stranded RNAi agent claim 1 , wherein the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length.

26. The double stranded RNAi agent of claim 1 , wherein at least one strand comprises a 3′ overhang of at least 2 nucleotides.

27. The double stranded RNAi agent of claim 1 , wherein substantially all of the nucleotides of the sense strand and substantially all of the nucleotides of the antisense strand are modified nucleotides.

28. The double stranded RNAi agent of claim 1 , wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand are modified nucleotides.

29. The double stranded RNAi agent of claim 9 , wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the 3′-terminus of one strand.

30. The double stranded RNAi agent of claim 9 , wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the 5′-terminus of one strand.

31. The double stranded RNAi agent of claim 9 , wherein the phosphorothioate or methylphosphonate internucleotide linkage is at both the 5′- and 3′-terminus of one strand.

32. The double stranded RNAi agent of claim 9 , wherein the double stranded RNAi agent comprises 6-8 phosphorothioate internucleotide linkages.

33. The double stranded RNAi agent of claim 7 , wherein the one or more GalNAc derivatives is conjugated to the 3′-terminus of the sense strand.

34. The double stranded RNAi agent of claim 33 , wherein the RNAi agent is conjugated to the ligand as shown in the following schematic

35. The dsRNA agent of claim 1 , wherein the antisense strand comprises a nucleotide seque

Assignees

• Alnylam Pharmaceuticals Inc

Inventors

• Fitzgerald Kevin
• Hinkle Gregory

Classifications

• A61P1/16

  for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics · CPC title

• C07H19/067

  with ribosyl as the saccharide radical · CPC title

• C12N2310/322

  2'-R Modification · CPC title

• C12N2310/3515

  Lipophilic moiety, e.g. cholesterol · CPC title

• C07H21/02

  with ribosyl as saccharide radical · CPC title