Therapeutic compositions
This application relates to therapeutic siRNA agents and methods of making and using the agents.
1. An RNA agent for inhibiting the expression of a target human gene in a cell, comprising a sense sequence and an antisense sequence, wherein the sense sequence has one or more asymmetrical 2′-O alkyl modifications and the antisense sequence has 4-20 phosphorothioate modifications, wherein the antisense sequence has fewer asymmetrical 2′-O alkyl modifications than the sense sequence, wherein the sense sequence comprises a conjugate group, and wherein the antisense sequence targets the human gene sequence.
2. The RNA agent of claim 1 , wherein at least one of the 2′-O-alkyl modifications is a 2′-OMe modification.
3. The RNA agent of claim 1 , wherein the sense sequence has 4-12 2′-O-alkyl modifications.
4. The RNA agent of claim 3 , wherein at least 4 of the asymmetrical 2′-O-alkyl modifications are within the 6 terminal nucleotides of the 5′ end or 3′ end of the sense sequence.
5. The RNA agent of claim 3 , wherein at least 4 of the asymmetrical 2′-O-alkyl modifications are within the 6 terminal nucleotides of the 5′ end or 3′ end of the sense sequence, and at least one of the 2′-O-alkyl modifications is in another portion of the sense sequence.
6. The RNA agent of claim 3 , wherein at least 4 of the asymmetrical 2′-O-alkyl modifications are within the 4 terminal nucleotides of the 5′ end or 3′ end of the sense sequence.
7. The RNA agent of claim 1 , wherein at least 4 of the phosphorothioate modifications are within the 4 terminal nucleotides of the 5′ end or 3′ end of the antisense sequence.
8. The RNA agent of claim 7 , wherein the sense sequence further comprises 2 phosphorothioate modifications within the 2 terminal nucleotides of the 5′ end or 3′ end of the sense sequence.
9. The RNA agent of claim 1 , wherein the antisense sequence has 6-20 phosphorothioate modifications.
10. The RNA agent of claim 1 , wherein the sense and antisense sequences of the RNA agent are fully complementary to each other.
11. The RNA agent of claim 1 , wherein the RNA agent is at least 21 nucleotides in length, and the duplex region of the RNA agent is about 19 nucleotides in length.
12. The RNA agent of claim 1 , wherein the RNA agent has a duplex region of about 19-21 nucleotides in length and one or two 3′ overhangs of about 2 nucleotides in length.
13. The RNA agent of claim 1 , wherein the sense sequence further comprises at least one asymmetric modification selected from the group consisting of 2′-5′-linkages, L sugars, modified sugars, nucleobase modifications, cation groups, Zwitterionic groups, and conjugate groups.
14. The RNA agent of claim 13 , wherein the modification is 2′-5′ linkages, and the 2′-5′ linkage is phosphorothioate.
15. The RNA agent of claim 13 , wherein the modification is L sugars, and the L sugar is L ribose or L-arabinose sugar.
16. The RNA agent of claim 13 , wherein the modification is modified sugars, and the modified sugar is a locked nucleic acid, hexose nucleic acid or cyclohexane nucleic acid.
17. The RNA agent of claim 1 , wherein the antisense sequence further comprises at least one asymmetric modification selected from the group consisting of 2′-5′-linkages, L sugars, modified sugars, nucleobase modifications, cation groups, Zwitterionic groups, and conjugate groups.
18. The RNA agent of claim 17 , wherein the modification is 2′-5′ linkages, and the 2′-5′ linkage is phosphorothioate.
19. The RNA agent of claim 17 , wherein the modification is a L sugar, and the L sugar is L ribose or L-arabinose sugar.
20. The RNA agent of claim 17 , wherein the modification is a modified sugar, and the modified sugar is a locked nucleic acid, a hexose nucleic acid, or a cyclohexane nucleic acid.