Short interfering RNA (siRNA) analogues
View Patent ↗The present invention is directed to novel double-stranded short interfering (siRNA) analogues comprising locked nucleic acid (LNA) monomers. Such compounds induces sequence-specific post-transcriptional gene silencing in many organisms by a process known as RNA interference (RNAi). The compounds disclosed herein has improved properties compared to non-modified siRNAs and may, accordingly, be useful as therapeutic agents, e.g., in the treatment of various cancer forms. More particularly, the present invention is directed to siRNA analogues comprising a sense strand and an antisense strand, wherein each strand comprises 12-35 nucleotides and wherein the siRNA analogues comprise at least one locked nucleic acid (LNA) monomer.
1. A double-stranded compound comprising a sense strand and an antisense strand, wherein each strand comprises 17-25 nucleotides, wherein at least one of the strands has a 3′ overhang and wherein said compound comprises at least one locked nucleic acid (LNA) monomer located at the 5′ end of the sense strand, the LNA monomer having the structure
wherein
X is selected from the group consisting of 0, S and NR″, where R″ is H or C 1-4 alkyl;
B is a nucleobase;
Z and Z* are independently absent or selected from the group consisting of an internucleoside linkage group, a terminal group and a protection group; so that when the LNA monomer is located at the 3′ end, Z is a terminal group and Z* is an internucleoside linkage group; when the LNA monomer is located at the 5′ end, Z is absent and Z* is a terminal group; and when the LNA monomer is located within the nucleotide sequence, Z is absent and Z* is an internucleoside linkage group.
2. The compound according to claim 1 , wherein the sense strand comprises 1-10 LNA monomers.
3. The compound according to claim 1 , wherein at least two LNA monomers are located at the 5′ end of the sense strand.
4. The compound according to claim 1 , wherein at least one LNA monomer is located at the 3′ end of the sense strand.
5. The compound according to claim 4 , wherein at least two LNA monomers are located at the 3′ end of the sense strand.
6. The compound according to claim 1 , wherein the antisense strand comprises at least one LNA monomer.
7. The compound according to claim 6 , wherein the antisense strand comprises 1-10 LNA monomers.
8. The compound according to claim 6 , wherein at least one LNA monomer is located at the 3′ end of the antisense strand.
9. The compound according to claim 8 , wherein at least two LNA monomers are located at the 3′ end of the antisense strand.
10. The compound according to claim 9 , wherein at least three LNA monomers are located at the 3′ end of the antisense strand.
11. The compound according to claim 1 , wherein no LNA monomer is located at the 5′ end of the antisense strand.
12. The compound according to claim 6 , wherein the sense strand comprises 1-10 LNA monomers and the antisense strand comprises 1-10 LNA monomers.
13. The compound according to claim 6 , wherein the sense strand comprises at least one LNA monomer at the 5′ end and at least one LNA monomer at the 3′ end, and wherein the antisense strand comprises at least one LNA monomer at the 3′ end.
14. The compound according to claim 6 , wherein the sense strand comprises at least one LNA monomer at the 5′ end and at least one LNA monomer at the 3′ end, and wherein the antisense strand comprises at least two LNA monomers at the 3′ end.
15. The compound according to claim 6 , wherein the sense strand comprises at least two LNA monomers at the 5′ end and at least two LNA monomers at the 3′ end, and wherein the antisense strand comprises at least two LNA monomers at the 3′ end.
16. The compound according to claim 6 , wherein the sense strand comprises at least two LNA monomers at the 5′ end and at least two LNA monomers at the 3′ end, and wherein the antisense strand comprises at least three LNA monomers at the 3′ end.
17. The compound according to claim 1 , wherein no LNA monomer is located at the 5′ end of the antisense strand.
18. The compound according to claim 1 , wherein the sense strand comprises at least one LNA monomer in at least one of the positions 9-13 counted from the 5′ end.
19. The compound according to claim 18 , wherein the sense strand comprises a LNA monomer in position 10.
20. The compound according to claim 18 , wherein the sense strand comprises a LNA monomer in position 11.
21. The compound according to claim 18 , wherein the sense strand comprises a LNA monomer in position 12.
22. The compound according to claim 1 , wherein each strand comprises 20-22 nucleotides.
23. The compound according to claim 1 , wherein X is selected from the group consisting of O, S and NH.
24. The compound according to claim 23 , wherein X is O.
25. The compound according to claim 1 , wherein said LNA monomer is in the beta-D form.
26. A pharmaceutical composition comprising the compound according to claim 1 and a pharmaceutically acceptable diluent, carrier or adjuvant.
27. The compound according to claim 19 wherein the nucleobase of the LNA monomer at position 10 is thymine (T).
28. The compound of any of claims 1 - 2 , 3 - 11 , 12 - 21 , 22 and 23 - 23 wherein the LNA is beta D-oxy LNA.