Apolipoprotein C3 (APOC3) iRNA compositions and methods of use thereof
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.
1. A double stranded RNAi agent for inhibiting expression of apolipoprotein C3 (APOC3) in a cell,
wherein the double stranded RNAi agent comprises a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand comprises at least 17 contiguous nucleotides from the nucleotide sequence of 5′-AAGGGACAGUAUUCUCAGUGC-3′ (SEQ ID NO:125), and wherein the antisense strand comprises at least 18 contiguous nucleotides from the nucleotide sequence of 5′-GCACUGAGAAUACUGUCCCUUUU-3′ (SEQ ID NO:228),
wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand are modified nucleotides;
wherein the sense strand comprises at least two 2′-fluoro modified nucleotides, at least seven 2′-O-methyl modified nucleotides, and at least two phosphorothioate internucleotide linkages;
wherein the antisense strand comprises at least six 2′-fluoro modified nucleotides, at least ten 2′-O-methyl modified nucleotides, at least one phosphorothioate internucleotide linkage at the 3′-terminus and at least one phosphorothioate internucleotide linkage at the 5′-terminus; and wherein:
at least two of the 2′-O-methyl modified nucleotides on the sense strand are 2′-O-methyl modified adenosine nucleotides at position 6 or position 8 of the sense strand counting from the 5′-end; or
at least one of the 2′-O-methyl modified nucleotides on the sense strand is a 2′-O-methyl modified uridine nucleotide at position 12 of the sense strand counting from the 5′-end; or
at least three of the 2′-fluoro modified nucleotides on the antisense strand are 2′-fluoro modified cytosine nucleotides at positions 2, 4, and 18 of the antisense strand counting from the 5′-end; or
at least three of the 2′-O-methyl modified nucleotides on the antisense strand are 2′-O-methyl modified adenosine nucleotides at positions 3, 7, and 9 of the antisense strand counting from the 5′-end; or
at least three of the 2′-O-methyl modified nucleotides on the antisense strand are 2′-O-methyl modified uridine nucleotides at positions 5, 11, and 21 of the antisense strand counting from the 5′-end; or
at least three of the 2′-O-methyl modified nucleotides on the antisense strand are 2′-O-methyl modified cytosine nucleotides at positions 13, 17, and 19 of the antisense strand counting from the 5′-end; and
wherein the sense strand is conjugated to a ligand.
2. The double stranded RNAi agent of claim 1 , wherein the sense strand comprises three 2′-fluoro modified nucleotides at positions 9, 10, 11 counting from the 5′-end.
3. The double stranded RNAi agent of claim 1 , wherein at least two of the 2′ O-methyl modified nucleotides on the sense strand are 2′-O-methyl modified guanosine nucleotides.
4. The double stranded RNAi agent of claim 3 , wherein at least one of the two 2′-O-methyl modified guanosine nucleotides on the sense strand is at position 4 or position 18 counting from the 5′-end.
5. The double stranded RNAi agent of claim 1 , wherein the sense strand comprises at least one abasic nucleotide.
6. The double stranded RNAi agent of claim 1 , wherein the double stranded region comprises a non-canonical base pair.
7. The double stranded RNAi agent of claim 1 , wherein each strand is 17-30 nucleotides in length.
8. The double stranded RNAi agent of claim 1 , wherein the RNAi agent is a double ended bluntmer of 21 nucleotides in length.
9. 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.
10. A pharmaceutical composition comprising the double stranded double stranded RNAi agent of claim 1 .
11. A double stranded RNAi agent for inhibiting expression of apolipoprotein C3 (APOC3) in a cell,
wherein the double stranded RNAi agent comprises a sense strand and an antisense strand forming a double-stranded region, wherein the sense strand comprises at least 17 contiguous nucleotides from the nucleotide sequence of 5′-AAGGGACAGUAUUCUCAGUGC-3′ (SEQ ID NO:125), and wherein the antisense strand comprises at least 18 contiguous nucleotides from the nucleotide sequence of 5′-GCACUGAGAAUACUGUCCCUUUU-3′ (SEQ ID NO:228), wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand are modified nucleotides;
wherein the sense strand comprises three 2′-fluoro modified nucleotides at positions 9, 10, 11 counting from the 5′-end, at least seven 2′-O-methyl modified nucleotides, at least one phosphorothioate internucleotide linkage at the 3′-terminus and at least one phosphorothioate internucleotide linkage at the 5′-terminus;
wherein at least two of the seven 2′-O-methyl modified nucleotides are 2′-O-methyl modified guanosine nucleotides, at least two of the seven 2′-O-methyl modified nucleotides are 2′-O-methyl modified adenosine nucleotides, and at least one of the seven 2′-O-methyl modified nucleotides is a 2′-O-methyl modified uridine nucleotide;
wherein the antisense strand comprises at least six 2′-fluoro modified nucleotides, at least ten 2′-O-methyl modified nucleotides, at least one phosphorothioate internucleotide linkage at the 3′-terminus and at least one phosphorothioate internucleotide linkage at the 5′-terminus,
wherein at least three of the six 2′-fluoro modified nucleotides are 2′-fluoro modified cytosine nucleotides, at least three of the ten 2′-O-methyl modified nucleotides are 2′-O-methyl modified adenosine nucleotides, at least three of the ten 2′-O-methyl modified nucleotides are 2′-O-methyl modified uridine nucleotides, and at least three of the ten 2′-O-methyl modified nucleotides are 2′-O-methyl modified cytosine nucleotides; and
wherein the sense strand is conjugated to a ligand.
12. The double stranded RNAi agent of claim 11 , wherein the sense strand comprises at least one abasic nucleotide.
13. The double stranded RNAi agent of claim 11 , wherein the double stranded region comprises a non-canonical base pair.
14. The double stranded RNAi agent of claim 11 , wherein each strand is 17-30 nucleotides in length.
15. The double stranded RNAi agent of claim 11 , wherein the RNAi agent is a double ended bluntmer of 21 nucleotides in length.
16. The double stranded RNAi agent of claim 11 , wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker.
17. A pharmaceutical composition comprising the double stranded double stranded RNAi agent of claim 11 .