Complement component C5 iRNA compositions and methods of use thereof
The invention relates to iRNA, e.g., double-stranded ribonucleic acid (dsRNA), compositions targeting the complement component C5 gene, and methods of using such iRNA, e.g., dsRNA, compositions to inhibit expression of C5 and to treat subjects having a complement component C5-associated disease, e.g., paroxysmal nocturnal hemoglobinuria.
1. A double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of complement component C5,
wherein said dsRNA comprises a sense strand and an antisense strand forming a double stranded region 15-30 nucleotide pairs in length,
wherein said sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from nucleotides 769-821 of SEQ ID NO:1 and said antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the corresponding nucleotide sequence of SEQ ID NO:5,
wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a nucleotide modification.
2. The dsRNA agent of claim 1 , wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from nucleotides 780-802 of SEQ ID NO:1.
3. The dsRNA agent of claim 1 , wherein at least one of said nucleotide modifications is selected from the group consisting of a 3′-terminal deoxy-thymine (dT) nucleotide modification, a 2′-O-methyl nucleotide modification, a 2′-fluoro nucleotide modification, a 2′-deoxy-nucleotide modification, a locked nucleotide modification, an abasic nucleotide modification, a 2′-amino-nucleotide modification, a 2′-alkyl-nucleotide modification, a morpholino nucleotide modification, a phosphoramidate modification, a non-natural base comprising nucleotide modification, a nucleotide comprising a 5′-phosphorothioate group modification, and a terminal nucleotide linked to a cholesteryl derivative or a dodecanoic acid bisdecylamide group modification.
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; or at least one strand comprises a 3′ overhang of at least 2 nucleotides.
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 6 , 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 8 , wherein the dsRNA agent is conjugated to the ligand as shown in the following schematic
and, wherein X is O or S.
11. The dsRNA agent of claim 1 , wherein the double-stranded region is 15-30 nucleotide pairs in length; 17-23 nucleotide pairs in length; 7-25 nucleotide pairs in length; 23-27 nucleotide pairs in length; 19-21 nucleotide pairs in length; or 21-23 nucleotide pairs in length.
12. The dsRNA agent of claim 1 , wherein each strand is 15-30 nucleotides in length; or 19-30 nucleotides in length.
13. The dsRNA agent of claim 1 , wherein the agent further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage.
14. An isolated cell containing the dsRNA agent of claim 1 .
15. A pharmaceutical composition for inhibiting expression of a complement component C5 gene comprising the dsRNA agent of claim 1 .
16. The dsRNA agent of claim 1 , wherein the nucleotide modifications are independently selected from the group consisting of 2′-O-methyl nucleotide modifications, 2′-fluoro nucleotide modifications, and deoxy-thymine (dT) nucleotide modifications.
17. The dsRNA agent of claim 13 , wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the 3′-terminus of one strand.
18. The dsRNA agent of claim 17 , wherein said strand is the antisense strand.
19. The dsRNA agent of claim 17 , wherein said strand is the sense strand.
20. The dsRNA agent of claim 13 , wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the 5′-terminus of one strand.
21. The dsRNA agent of claim 20 , wherein said strand is the antisense strand.
22. The dsRNA agent of claim 20 , wherein said strand is the sense strand.
23. The dsRNA agent of claim 13 , wherein the phosphorothioate or methylphosphonate internucleotide linkage is at both the 5′- and 3′-terminus of one strand.
24. The dsRNA agent of claim 23 , wherein said strand is the antisense strand.
25. The dsRNA agent of 49, wherein the dsRNA agent comprises 6-8 phosphorothioate internucleotide linkages.
26. The dsRNA agent of claim 25 , 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.
27. The dsRNA agent of claim 1 , wherein the antisense strand comprises at least 17 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of 5′-UAUUAUAAAAAUAUCUUGCUUUU-3′ (SEQ ID NO:113).
28. The dsRNA agent of claim 1 , wherein the sense strand comprises at least 17 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of 5′-AAGCAAGAUAUUUUUAUAAUA-3′ (SEQ ID NO:62), and the antisense strand comprises at least 17 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of 5′-UAUUAUAAAAAUAUCUUGCUUUU-3′ (SEQ ID NO:113).
29. The dsRNA agent of claim 1 , wherein the antisense strand comprises the nucleotide sequence of 5′-UAUUAUAAAAAUAUCUUGCUUUU-3′ (SEQ ID NO:113).
30. The dsRNA agent of claim 1 , wherein the sense strand consists of the nucleotide sequence of 5′-AAGCAAGAUAUUUUUAUAAUA-3′ (SEQ ID NO:62), and the antisense strand consists of the nucleotide sequence of 5′-UAUUAUAAAAAUAUCUUGCUUUU-3′ (SEQ ID NO:113).
31. The dsRNA agent of claim 30 , wherein the sense strand consists of 5′-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3′ (SEQ ID NO:2876) and the antisense strand consists of 5′-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3′ (SEQ ID NO:2889),
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; Af, Gf, Cf and Uf are 2′-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.
32. The dsRNA agent of claim 31 , wherein the sense and antisense strands consist of the nucleotide sequences selected from the group consisting of 5′-AfaGfcAfaGfaUfAfUfulifuUfaUfaAfuAf-3′ (SEQ ID NO:122) and 5′-uAfuUfaUfaAfaAfauaUfcUfuGfcUfusUfsu-3′ (SEQ ID NO:173); 5′-AfsasGfcAfaGfaUfAffifulifuUfaUfaAfuAf-3′ (SEQ ID NO:164) and 5′-usAfsuUfaUfaAfaAfauaUfcUfuGfcUfususu-3′ (SEQ ID NO:215); 5′-asasGfcAfaGfaUfAffifulifuuAfuAfaua-3′ (SEQ ID NO:2875) and 5′-usAfsUfuAfuAfAfaAfauaUfcUfuGfcuususudTdT-3′ (SEQ ID NO:2888); 5′-asasGfcAfaGfaUfAfUfuUfuuAfuAfaua-3′ (SEQ ID NO: 2876) and 5′-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3′ (SEQ ID NO:2889); 5′-asasGfcAfaGfaUfAffifulifuuAfuaAfua-3′ (SEQ ID NO:2878) and 5′-usAfsUfuAfuAfAfaAfauaUfcUfuGfcuususudTdT-3′ (SEQ ID NO:2891); 5′-asasGfcAfaGfaUfAffifulifuuAfuaAfua-3′ (SEQ ID NO:2879) and 5′-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3′ (SEQ ID NO:2892); 5′-asasgcaagaUfaUfuuuuauaaua-3′ (SEQ ID NO:2884) and 5′-usAfsUfuAfuAfAfaAfauaUfcUfuGfcuususudTdT-3′ (SEQ ID NO:2897); and 5′-asasgcaagaUfaUfuuuuauaaua-3′ (SEQ ID NO:2885) and 5′-usAfsUfuAfuaAfaAfauaUfcUfuGfcuususudTdT-3′ (SEQ ID NO:2898),
wherein a, g, c and u are 2′-O-methyl (2′OMe) A, G, C, and U, respectively; Af, Gg, Cf and Uf are 2′-fluoro A, G, C and U, respectively; dT is a deoxy-thymine nucleotide; and s is a phosphorothioate linkage.