Antisense design

Patent metadata

Field	Value
Publication number	US-9428534-B2
Application number	US-201313797913-A
Country	US
Kind code	B2
Filing date	Mar 12, 2013
Priority date	Nov 18, 2002
Publication date	Aug 30, 2016
Grant date	Aug 30, 2016

Abstract

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.

First claim

The invention claimed is:

1. A gapmer oligomer having 10-15 nucleotide units wherein a segment of 8-10 DNA nucleotides is flanked by locked nucleotides, and wherein at least one or all of the internucleotide linkages is a phosphorothioate linkage.

2. The gapmer oligomer according to claim 1 , wherein the locked nucleotides have the formula

wherein
X represents O, S, or N—R;
R is selected from the group consisting of H and C 1 -C 6 alkyl, and C 1 -C 6 alkyl substituted with one or more substituents selected from the group consisting of halogen, nitro, C 1-6 alkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, and halogenated C 1-6 alkoxy;
n is 1;
(CH 2 ) n is optionally substituted with methyl; and
Base is independently selected from the group consisting of cytosine, methyl cytosine, uracil, thymine, adenine and guanine.

3. The gapmer oligomer of claim 1 wherein the flanks independently have 1-4 locked nucleotides.

4. The gapmer oligomer of claim 2 , wherein the flanks independently have 1-4 locked nucleotides.

5. The gapmer oligomer of claim 1 , wherein the flanks independently have 2-3 locked nucleotides.

6. The gapmer oligomer of claim 2 , wherein the flanks independently have 2-3 locked nucleotides.

7. The gapmer oligomer according to claim 3 , wherein the oligomer has 12-14 nucleotide units.

8. The gapmer oligomer according to claim 4 , wherein the oligomer has 12-14 nucleotide units.

9. The gapmer oligomer according to claim 5 , wherein the oligomer has 12-14 nucleotide units.

10. The gapmer oligomer according to claim 6 , wherein the oligomer has 12-14 nucleotide units.

11. The gapmer oligomer according to claim 1 , wherein the central region is 8-10 DNA nucleotides and the flanks are both independently 1-4 locked nucleotides, and wherein all of the internucleotide linkages is a phosphorothioate linkage.

12. The gapmer oligomer according to claim 1 , wherein the central region is 8-10 DNA nucleotides and the flanks are both independently 2-3 locked nucleotides, and wherein all of the internucleotide linkages is a phosphorothioate linkage.

13. The gapmer oligomer according to claim 11 , wherein the oligomer has 12-14 nucleotide units.

14. The gapmer oligomer according to claim 12 , wherein the oligomer has 12-14 nucleotide units.

15. The gapmer oligomer according to claim 14 , wherein the locked nucleotides have the formula

wherein
X represents O, S, or N—R;
R is selected from the group consisting of H and C 1 -C 6 alkyl, and C 1 -C 6 alkyl substituted with one or more substituents selected from the group consisting of halogen, nitro, C 1-6 alkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, and halogenated C 1-6 alkoxy;
n is 1;
(CH 2 ) n is optionally substituted with methyl; and
Base is independently selected from the group consisting of cytosine, methyl cytosine, uracil, thymine, adenine and guanine.

16. The gapmer oligomer of any of claims 1 wherein (CH 2 )n is substituted with methyl.

17. The gapmer oligomer of claim 2 wherein (CH 2 )n is substituted with methyl.

18. The gapmer oligomer of claim 3 wherein (CH 2 )n is substituted with methyl.

19. The gapmer oligomer of claim 4 wherein (CH 2 )n is substituted with methyl.

20. The gapmer oligomer of claim 5 wherein (CH 2 )n is substituted with methyl.

21. The gapmer oligomer of claim 6 wherein (CH 2 )n is substituted with methyl.

22. The gapmer oligomer of claim 7 wherein (CH 2 )n is substituted with methyl.

23. The gapmer oligomer of claim 8 wherein (CH 2 )n is substituted with methyl.

24. The gapmer oligomer of claim 9 wherein (CH 2 )n is substituted with methyl.

25. The gapmer oligomer of claim 10 wherein (CH 2 )n is substituted with methyl.

26. The gapmer oligomer of claim 11 wherein (CH 2 )n is substituted with methyl.

27. The gapmer oligomer of claim 12 wherein (CH 2 )n is substituted with methyl.

28. The gapmer oligomer of claim 13 wherein (CH 2 )n is substituted with methyl.

29. The gapmer oligomer of claim 14 wherein (CH 2 )n is substituted with methyl.

30. The gapmer oligomer of claim 15 wherein (CH 2 )n is substituted with methyl.

31. The gapmer oligomer of claim 1 wherein (CH 2 )n is unsubstituted.

32. The gapmer oligomer of claim 2 wherein (CH 2 )n is unsubstituted.

33. The gapmer oligomer of claim 3 wherein (CH 2 )n is unsubstituted.

34. The gapmer oligomer of claim 4 wherein (CH 2 )n is unsubstituted.

35. The gapmer oligomer of claim 5 wherein (CH 2 )n is unsubstituted.

36. The gapmer oligomer of claim 6 wherein (CH 2 )n is unsubstituted.

37. The gapmer oligomer of claim 7 wherein (CH 2 )n is unsubstituted.

38. The gapmer oligomer of claim 8 wherein (CH 2 )n is unsubstituted.

39. The gapmer oligomer of claim 9 wherein (CH 2 )n is unsubstituted.

40. The gapmer oligomer of claim 10 wherein (CH 2 )n is unsubstituted.

41. The gapmer oligomer of claim 11 wherein (CH 2 )n is unsubstituted.

42. The gapmer oligomer of claim 12 wherein (CH 2 )n is unsubstituted.

43. The gapmer oligomer of claim 13 wherein (CH 2 )n is unsubstituted.

44. The gapmer oligomer of claim 14 wherein (CH 2 )n is unsubstituted.

45. The gapmer oligomer of claim 15 wherein (CH 2 )n is unsubstituted.

46. The gapmer oligomer of claim 1 wherein the locked nucleotides have the formula:

wherein
X represents O;
n is 1;
(CH 2 )n is substituted with methyl; and
Base is independently selected from the group consisting of cytosine, methyl cytosine, uracil, thymine, adenine and guanine.

47. The gapmer oligomer of claim 46 , wherein the flanks independently have 1-4 locked nucleotides.

48. The gapmer oligomer of claim 46 , wherein the flanks independently have 2-3 locked nucleotides.

49. The gapmer oligomer according to claim 46 , wherein the oligomer has 12-14 nucleotide units.

50. The gapmer oligomer according to claim 47 , wherein the oligomer has 12-14 nucleotide units.

51. The gapmer oligomer according to claim 48 , wherein the oligomer has 12-14 nucleotide units.

52. The gapmer oligomer according to claim 46 , wherein the central region is 8-10 DNA nucleotides and the flanks are both independently 1-4 locked nucleotides, and wherein all of the internucleotide linkages is a phosphorothioate linkage.

53. The gapmer oligomer according to claim 46 , wherein the central region is 8-10 DNA nucleotides and the flanks are both independently 2-3 locked nucleotides, and wherein all of the internucleotide linkages is a phosphorothioate linka

Assignees

• Santaris Pharma As
• Roche Innovation Ct Copenhagen As

Inventors

• Christensen Signe M
• Mikkelsen Nikolaj Dam
• Frieden Miriam
• Hansen Henrik Frydenlund
• Koch Troels
• Pedersen Daniel Sejer
• Rosenbohm Christoph
• Thrue Charlotte Albaek
• Westergaard Majken

Classifications

• C12N15/111

  General methods applicable to biologically active non-coding nucleic acids · CPC title

• C07H21/04

  with deoxyribosyl as saccharide radical · CPC title

• C07H19/04

  Heterocyclic radicals containing only nitrogen atoms as ring hetero atom · CPC title

• A61K31/7088

  Compounds having three or more nucleosides or nucleotides · CPC title

• C12N15/1137

  against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138) · CPC title