Antisense design
A novel class of pharmaceuticals which comprises a Locked Nucleic Acid (LNA) which can be used in antisense therapy. These novel oligonucleotides have improved antisense properties. The novel oligonucleotides are composed of at least one LNA selected from beta-D-thio/amino-LNA or alpha-L-oxy/thio/amino-LNA. The oligonucleotides comprising LNA may also include DNA and/or RNA nucleotides.
1. A gapmer oligomer having 12-18 nucleotide units comprising a central region of at least 6 contiguous DNA nucleotides which is flanked both 5′ and 3′ by locked nucleotides of formula:
wherein X is O; (CH 2 )n is substituted with a C 1-6 alkyl; n is 1; and Base is independently selected from the group consisting of cytosine, methyl cytosine, uracil, thymine, adenine and guanine.
2. The gapmer oligomer of claim 1 wherein all of the locked nucleotides have the formula:
wherein (CH 2 )n is substituted with a methyl group.
3. The gapmer oligomer of claim 1 wherein at least one of the internucleotide linkages is a phosphorothioate linkage.
4. The gapmer oligomer of claim 1 wherein all of the internucleotide linkages are phosphorothioate linkages.
5. The gapmer oligomer according to claim 2 wherein at least one of the internucleotide linkages is a phosphorothioate linkages.
6. The gapmer oligomer according to claim 2 wherein all of the internucleotide linkages are phosphorothioate linkages.
7. The gapmer oligomer of claim 1 wherein in the central region is 8-12 DNA nucleotides.
8. The gapmer oligomer of claim 2 wherein in the central region is 8-12 DNA nucleotides.
9. The gapmer oligomer of claim 1 , wherein the central region is flanked both 5′ and 3′ by 2-6 locked nucleotides.
10. The gapmer oligomer of claim 2 , wherein the central region is flanked both 5′ and 3′ by 2-6 locked nucleotides.
11. The gapmer oligomer of claim 1 , wherein the central region is flanked both 5′ and 3′ by 3 or 4 locked nucleotides.
12. The gapmer oligomer of claim 2 , wherein the central region is flanked both 5′ and 3′ by 3 or 4 locked nucleotides.
13. The gapmer oligomer according to claim 1 , wherein the oligomer is 15, 16 or 17 nucleotide units in length.
14. The gapmer oligomer according to claim 2 , wherein the oligomer is 15, 16 or 17 nucleotide units in length.
15. The gapmer oligomer according to claim 1 , wherein all of the nucleotide units other than the DNA nucleotides of the central region are locked nucleotides.
16. The gapmer oligomer according to claim 2 , wherein all of the nucleotide units other than the DNA nucleotides of the central region are locked nucleotides.
17. The gapmer oligomer according to claim 1 , wherein at least one of the locked nucleotides has the formula
wherein (CH 2 )n is substituted with a C 1-6 alkyl; n is 1; and Base is independently selected from the group consisting of cytosine, methyl cytosine, uracil, thymine, adenine and guanine.
18. The gapmer oligomer of claim 17 wherein (CH 2 )n is substituted with a methyl group.
19. The gapmer oligomer of claim 17 wherein at least one of the internucleotide linkages is a phosphorothioate linkage.
20. The gapmer oligomer of claim 17 wherein all of the internucleotide linkages are phosphorothioate linkages.
21. The gapmer oligomer of claim 17 wherein the central region is 8-12 DNA nucleotides.
22. The gapmer oligomer of claim 17 , wherein the central region is flanked both 5′ and 3′ by 2-6 locked nucleotides.
23. The gapmer oligomer of claim 17 , wherein the central region is flanked both 5′ and 3′ by 3 or 4 locked nucleotides.
24. The gapmer oligomer of claim 17 , wherein all of the locked nucleotides have the formula
wherein (CH 2 )n is substituted with a C 1-6 alkyl; n is 1 and Base is independently selected from the group consisting of cytosine, methyl cytosine, uracil, thymine, adenine and guanine.
25. The gapmer oligomer of claim 17 , wherein the central region consists of 8-12 DNA nucleotides and is flanked both 5′ and 3′ by 2-6 locked nucleotides and wherein all of the internucleotide linkages are phosphorothioate linkages.
26. The gapmer oligomer according to claim 17 , wherein the oligomer is 15, 16 or 17 nucleotide units in length.
27. The gapmer oligomer according to claim 17 , wherein all of the nucleotide units other than the DNA nucleotides of the central region are locked nucleotides.
28. The gapmer oligomer of claim 18 wherein at least one of the internucleotide linkages is a phosphorothioate linkage.
29. The gapmer oligomer of claim 18 wherein all of the internucleotide linkages are phosphorothioate linkages.
30. The gapmer oligomer of claim 18 wherein in the central region is 8-12 DNA nucleotides.
31. The gapmer oligomer of claim 18 , wherein the central region is flanked both 5′ and 3′ by 2-6 locked nucleotides.
32. The gapmer oligomer of claim 18 , wherein the central region is flanked both 5′ and 3′ by 3 or 4 locked nucleotides.
33. The gapmer oligomer of claim 18 , wherein all of the locked nucleotides have the formula: formula
wherein (CH 2 )n is substituted with a methyl; n is 1; and Base is independently selected from the group consisting of cytosine, methyl cytosine, uracil, thymine, adenine and guanine.
34. The gapmer oligomer of claim 18 , wherein the central region consists of 8-12 DNA nucleotides and is flanked both 5′ and 3′ by 2-6 locked nucleotides and wherein all of the internucleotide linkages are phosphorothioate linkages.
35. The gapmer oligomer according to claim 18 , wherein the oligomer is 15, 16 or 17 nucleotide units in length.
36. The gapmer oligomer according to claim 18 or claim 34 , wherein all of the nucleotide units other than the DNA nucleotides of the central region are locked nucleotides.