Apolipoprotein C3 (APOC3) iRNA compositions and methods of use thereof

Patent metadata

Field	Value
Publication number	US-10407679-B2
Application number	US-201715597225-A
Country	US
Kind code	B2
Filing date	May 17, 2017
Priority date	Nov 17, 2014
Publication date	Sep 10, 2019
Grant date	Sep 10, 2019

Abstract

The present invention relates to RNAi agents, e.g., double-stranded RNAi agents, targeting the apolipoprotein C3 (APOC3) gene, and methods of using such RNAi agents to inhibit expression of APOC3 and methods of treating subjects having an APOC3 associated disorder, e.g., hypertriglyceridemia.

First claim

We claim:

1. A double stranded RNAi agent for inhibiting expression of apolipoprotein C3 (APOC3) in a cell, wherein said double stranded RNAi agent comprises a sense strand and an antisense strand forming a double-stranded region, wherein the antisense strand comprises the nucleotide sequence 5′-UGAAUACUGUCCCUUUUAAGCAA-3′ (SEQ ID NO:670),
wherein substantially all of the nucleotides of at least one strand are modified nucleotides selected from the group consisting of a 2′-O-methyl modified nucleotide, a 2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, a nucleotide comprising a 5′ phosphate or 5′ phosphate mimic, a nucleotide comprising vinyl phosphate, a nucleotide comprising adenosine-glycol nucleic acid (GNA), and a nucleotide comprising thymidine-glycol nucleic acid (GNA) S-Isomer, and
wherein said sense strand is conjugated to a ligand attached at the 3′-terminus.

2. The double stranded RNAi agent of claim 1 , further comprising at least one modified nucleotide selected from the group consisting of a 3′-terminal deoxy-thymine (dT) 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, a 2′-C-alkyl-modified nucleotide, a 2′-hydroxly-modified nucleotide, a 2′-methoxyethyl modified nucleotide, a 2′-O-alkyl-modified nucleotide, a morpholino nucleotide, a nucleoside 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 5′-phosphorothioate group, a nucleotide comprising a 5′-methylphosphonate group, a nucleotide comprising 2-hydroxymethyl-tetrahydrofurane-5-phosphate, a nucleotide comprising 2′-deoxythymidine-3′phosphate, a nucleotide comprising 2′-deoxyguanosine-3′-phosphate, and a terminal nucleotide linked to a cholesteryl derivative or a dodecanoic acid bisdecylamide group.

3. The double stranded RNAi agent of claim 1 , wherein the modified nucleotides are 2′-O-methyl modified nucleotides and 2′-fluoro modified nucleotides.

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

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

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

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

8. The double stranded RNAi agent of claim 1 , wherein the double-stranded region is 15-30 nucleotide pairs in length.

9. The double stranded RNAi agent of claim 1 , wherein the double-stranded region is 17-25 nucleotide pairs in length.

10. The double stranded RNAi agent of claim 1 , wherein the double-stranded region is 21-23 nucleotide pairs in length.

11. The double stranded RNAi agent of claim 1 , wherein each strand is 23-30 nucleotides in length.

12. The double stranded RNAi agent of claim 1 , wherein each strand is 25-30 nucleotides in length.

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

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

15. The double stranded RNAi agent of claim 14 , wherein the double stranded RNAi agent comprises 6-8 phosphorothioate internucleotide linkages.

16. The double stranded RNAi of claim 15 , wherein the antisense strand comprises two phosphorothioate internucleotide linkages at the 5′-terminus and two phosphorothioate internucleotide linkages at the 3′-terminus, and the sense strand comprises at least two phosphorothioate internucleotide linkages at either the 5′-terminus or the 3′-terminus.

17. The double stranded RNAi agent of claim 1 , wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker.

18. The double stranded RNAi agent of claim 1 , wherein the ligand is

19. The double stranded RNAi agent of claim 1 , wherein the RNAi agent is conjugated to the ligand as shown in the following schematic

wherein X is 0 or S.

20. The double stranded RNAi agent of claim 1 , wherein the sense strand comprises the nucleotide sequence 5′-GCUUAAAAGGGACAGUAUUCA-3′ (SEQ ID NO:659) and the antisense strand comprises the nucleotide sequence 5′-UGAAUACUGUCCCUUUUAAGCAA-3′ (SEQ ID NO:670).

21. The double stranded RNAi agent of claim 20 , wherein the sense strand comprises the nucleotide sequence 5′-gscsuuaaAfaGfGfGfacaguauuca-3′ (SEQ ID NO:680) and the antisense strand comprises the nucleotide sequence 55′-usGfsaauAfcUfGfucccUfuUfuaagcsasa-3′ (SEQ DI NO:691), wherein a, c, g, and u are 2′-O-methyl (2′-OMe) A, C, G, and U; Af, Cf, Gf and Uf are 2′-fluoro A, C, G and U; and s is a phosphorothioate linkage.

22. The double stranded RNAi agent of claim 20 , wherein the sense strand comprises the nucleotide sequence 5′-gscsuuaaAfaGfGfGfacaguauuca-3′ (SEQ ID NO:681) and the antisense strand comprises the nucleotide sequence 5′-usGfsaauacugucccUfuuuaagcsasa-3′ (SEQ ID NO:692),
wherein a, c, g, and u are 2′-O-methyl (2′-OMe) A, C, G, and U; Af, Cf, Gf and Uf are 2′-fluoro A, C, G and U; and s is a phosphorothioate linkage.

23. The double stranded RNAi agent of claim 20 , wherein the sense strand comprises the nucleotide sequence 5′-gscsuuaaaaGfgGfacaguauuca-3′ (SEQ ID NO:738) and the antisense strand comprises the nucleotide sequence 5′-usGfsaauacugucCfcUfuuuaagcsasa-3′ (SEQ ID NO:749),
wherein a, c, g, and u are 2′-O-methyl (2′-OMe) A, C, G, and U; Af, Cf, Gf and Uf are 2′-fluoro A, C, G and U; and s is a phosphorothioate linkage.

24. The double stranded RNAi agent of claim 20 , wherein the sense strand comprises the nucleotide sequence 5′-gscsuuaaaaGfgGfacagu(Agn)uuca-3′ (SEQ ID NO:744) and the antisense strand comprises the nucleotide sequence 5′-usGfsaauacugucCfcUfuuuaagcsasa-3′ (SEQ ID NO:755),
wherein a, c, g, and u are 2′-O-methyl (2′-OMe) A, C, G, and U; Af, Cf, Gf and Uf are 2′-fluoro A, C, G and U; (Agn) is adenosine-glycol nucleic acid (GNA); and s is a phosphorothioate linkage.

25. The double stranded RNAi agent of claim 20 , wherein the sense strand comprises the nucleotide sequence 5′-gscsuuaaAfaGfGfGfacaguauucaL96-3′ (SEQ ID NO:680) and the antisense strand comprises the nucleotide sequence 55′-usGfsaauAfcUfGfucccUfuUfuaagcsasa-3′ (SEQ DI NO:691),
wherein a, c, g, and u are 2′-O-methyl (2′-OMe) A, C, G, and U; Af, Cf, Gf and Uf are 2′-fluoro A, C, G and U; s is a phosphorothioate linkage; and L96 is N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol.

26. The double stranded RNAi agent of claim 20 , wherein the sense strand comprises the nucleotide sequence 5′-gscsuuaaAfaGfGfGfacaguauucaL96-3′ (SEQ ID NO:681)
and the antisense str

Assignees

• Alnylam Pharmaceuticals Inc

Inventors

• Fitzgerald Kevin
• Querbes William
• Butler James
• Williams Stephanie
• Liebow Abigail
• Hinkle Gregory
• Maier Martin
• Milstein Stuart
• Kuchimanchi Satyanarayana
• Manoharan Muthiah

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P9/12

  Antihypertensives · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P3/06

  Antihyperlipidemics · CPC title