Short interfering RNA (siRNA) analogues
The present invention is directed to novel double-stranded short interfering (siRNA) analogs 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 analogs comprising a sense strand and an antisense strand, wherein each strand comprises 12-35 nucleotides and wherein the siRNA analogs 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 12-35 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.
2. The compound according to claim 1 , wherein at least three LNA monomers are located at the 3′ end of the antisense strand.
3. The compound according to claim 1 , wherein no LNA monomer is located at the 5′ end of the antisense strand.
4. The compound according to claim 1 , wherein the sense strand comprises 1-10 LNA monomers and the antisense strand comprises 1-10 LNA monomers.
5. The compound according to claim 1 , 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.
6. The compound according to claim 1 , 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.
7. The compound according to claim 1 , 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.
8. The compound according to claim 1 , 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.
9. The compound according to claim 4 , wherein no LNA monomer is located at the 5′ end of the antisense strand.
10. The compound according to claim 1 , wherein the sense strand comprises at least one LNA monomer in at least one of positions 9-13 counted from the 5′ end.
11. The compound according to claim 1 , wherein the sense strand comprises a LNA monomer in position 10.
12. The compound according to claim 1 , wherein the sense strand comprises a LNA monomer in position 11.
13. The compound according to claim 1 , wherein the sense strand comprises a LNA monomer in position 12.
14. The compound according to claim 1 , wherein each strand comprises 20-22 nucleotides.
15. The compound according to claim 1 , wherein the LNA monomer is in the beta-D form.
16. A pharmaceutical composition comprising the compound according to claim 1 and a pharmaceutically acceptable diluent, carrier or adjuvant.
17. The compound according to claim 1 wherein the nucleobase of the LNA monomer at position 10 is thymine (T).
18. The compound according to claim 1 wherein at least one 3′ overhang is 1-3 nucleotides in length.
19. The compound according to claim 1 wherein the at least one 3′ overhang is 1-3 nucleotides in length, and wherein there is no LNA monomer located at the 5′ end of the antisense strand.
20. The compound according to claim 1 wherein the at least one 3′ overhang is 1-3 nucleotides in length, wherein there is no LNA monomer located at the 5′ end of the antisense strand, and wherein there is at least one LNA monomer at the 5′ end of the sense strand.
21. The compound according to claim 1 , wherein both the sense strand and the antisense strand comprise a 3′ overhang.
22. The compound according to claim 1 , wherein both the sense strand and the antisense strand comprise a 3′ overhang and wherein each 3′ overhang is, independently, 1-3 nucleotides in length.
23. The compound according to claim 1 , wherein both the sense strand and the antisense strand comprise a 3′ overhang, and wherein each 3′ overhang is 3 nucleotides in length.
24. The compound according to claim 1 , wherein both the sense strand and the antisense strand comprise a 3′ overhang, wherein each 3′ overhang is, independently, 1-3 nucleotides in length, and wherein there is no LNA monomer located at the 5′ end of the antisense strand.
25. The compound according to claim 1 , wherein both the sense and antisense strand comprise a 3′ overhang, wherein each 3′ overhang is, independently, 1-3 nucleotides in length, wherein there is no LNA monomer located at the 5′ end of the antisense strand, and wherein there is at least one LNA monomer at the 5′ end of the sense strand.
26. The compound according to claim 1 , wherein both the sense strand and the antisense strand comprise a 3′ overhang, wherein each 3′ overhand is, independently, 1-3 nucleotides in length, wherein there is no LNA monomer located at the 5′ end of the antisense strand, and wherein at least two LNA monomers are located at the 3′ end of the antisense strand.
27. The compound according to claim 1 , wherein both the sense strand and antisense strand comprise a 3′ overhang, wherein each 3′ overhang is, independently, 1-3 nucleotides in length, and wherein there is no LNA monomer located at the 5′ end of the antisense strand, wherein the antisense strand comprises at least one LNA monomer at the 3′ end, and wherein the sense strand comprises at least one LNA monomer at the 5′ end and at least one LNA monomer at the 3′ end.
28. The compound according to claim 1 , wherein both the sense strand and the antisense strand comprise a 3′ overhang, wherein each 3′ overhang is, independently, 1-3 nucleotides in length, wherein there is no LNA monomer located at the 5′ end of the antisense strand, wherein the antisense strand comprises at least one LNA monomer at the 3′ end, 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 sense strand comprises at least one LNA monomer in at least one of positions 9-13 counted from the 5′ end.
29. The compound according to claim 1 , wherein both the sense and antisense strand comprise a 3′ overhang, wherein each 3′ overhang is, independently, 3 nucleotides in length, wherein there is no LNA monomer located at the 5′ end of the antisense strand, wherein the antisense strand comprises at least one LNA monomer at the 3′ end, wherein the sense strand comprises at least one LNA monomer at the 5′ end and at least one LNA monomer at the 3′ end.
30. The compound according to claim 1 , wherein both the sense and antisense strand comprise a 3′ overhang, wherein each 3′ overhang is, independently, 3 nucleotides in length, wherein there is no LNA monomer located at the 5′ end of the antisense strand, wherein the antisense strand comprises at least one LNA monomer at the 3′ end, 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 sense strand comprises at least one LNA monomer at least one of positions 9-13 counted from the 5′ end.