Antisense design

Patent metadata

Field	Value
Publication number	US-9951333-B2
Application number	US-201715618892-A
Country	US
Kind code	B2
Filing date	Jun 9, 2017
Priority date	Nov 18, 2002
Publication date	Apr 24, 2018
Grant date	Apr 24, 2018

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 oligonucleotide, 10-30 nucleotides in length, comprising a nucleotide of formula:

wherein X is O; one of the H in (CH 2 )n is replaced with a C 1-6 alkyl substituent; n is 1; and Base is independently selected from the group consisting of cytosine, methyl cytosine, uracil, thymine, adenine and guanine, wherein the gapmer oligonucleotide further comprises a 2′ substituted non-locked nucleotide wherein the 2′ substituent is selected from the group consisting of halogen and C 1 -C 9 alkoxy.

2. The gapmer oligonucleotide according to claim 1 , wherein one of the H in (CH 2 )n is replaced with methyl.

3. The gapmer oligonucleotide according to claim 1 , wherein the 2′ substituent is 2′-fluoro.

4. The gapmer oligonucleotide according to claim 1 , wherein the 2′ substituent is C 1 -C 9 alkoxy.

5. The gapmer oligonucleotide according to claim 1 , wherein the 2′ substituent is C 1 -C 9 methoxy or ethoxy.

6. The gapmer oligonucleotide according to claim 2 , wherein the 2′ substituent is 2′-fluoro.

7. The gapmer oligonucleotide according to claim 2 , wherein the 2′ substituent is C 1 -C 9 alkoxy.

8. The gapmer oligonucleotide according to claim 2 , wherein the 2′ substituent is C 1 -C 9 methoxy or ethoxy.

9. The gapmer oligonucleotide according to claim 1 , wherein the gapmer oligonucleotide is 12-18 nucleotide in length.

10. A gapmer oligonucleotide, 10-30 nucleotides in length, comprising a gap region of at least 6 contiguous DNA units flanked 5′ and 3′ by 2′-O modified nucleotides, wherein at least one of the nucleotides in the flanks is a 2′-O modified nucleotide of formula

wherein X is O; one of the H in (CH 2 )n is replaced with a C 1-6 alkyl substituent; n is 1; and Base is independently selected from the group consisting of cytosine, methyl cytosine, uracil, thymine, adenine and guanine, and at least one of the nucleotides in the flanks is a 2′ substituted non-locked nucleotide wherein the 2′ substituent is selected from the group consisting of halogen or C 1 -C 9 alkoxy.

11. The gapmer oligonucleotide according to claim 10 , wherein one of the H in (CH 2 )n is replaced with methyl.

12. The gapmer oligonucleotide according to claim 10 , wherein the 2′ substituent is 2′-fluoro.

13. The gapmer oligonucleotide according to claim 10 , wherein the 2′ substituent is C 1 -C 9 alkoxy.

14. The gapmer oligonucleotide according to claim 10 , wherein the 2′ substituent is C 1 -C 9 methoxy.

15. The gapmer oligonucleotide according to claim 10 , wherein the 2′ substituent is 2′-fluoro.

16. The gapmer oligonucleotide according to claim 11 , wherein the 2′ substituent is C 1 -C 9 alkoxy.

17. The gapmer oligonucleotide according to claim 11 , wherein the 2′ substituent is C 1 -C 9 methoxy.

18. The gapmer oligonucleotide according to claim 10 , wherein at least one of the internucleoside linkages is a phosphorothioate linkage.

19. The gapmer oligonucleotide of claim 10 , wherein all of the internucleotide linkages are phosphorothioate linkages.

20. The gapmer oligonucleotide according to claim 11 , wherein at least one of the internucleotide linkages is a phosphorothioate linkages.

21. The gapmer oligonucleotide according to claim 11 , wherein all of the internucleotide linkages are phosphorothioate linkages.

22. The gapmer oligonucleotide of claim 10 , wherein in the central region is 8-12 DNA nucleotides.

23. The gapmer oligonucleotide of claim 11 , wherein in the central region is 8-12 DNA nucleotides.

24. The gapmer oligonucleotide of claim 10 , wherein the central region is flanked both 5′ and 3′ by 1-6 2′-O modified nucleotides.

25. The gapmer oligonucleotide of claim 11 , wherein the central region is flanked both 5′ and 3′ by 1-6 2′-O modified locked nucleotides.

26. The gapmer oligonucleotide of claim 10 , wherein the central region is flanked both 5′ and 3′ by locked nucleotides.

27. The gapmer oligonucleotide of claim 11 , wherein the central region is flanked both 5′ and 3′ by locked nucleotides.

28. The gapmer oligonucleotide according to claim 10 , wherein the oligomer is 12-18 nucleotides units in length.

29. The gapmer oligonucleotide according to claim 11 , wherein the oligomer is 12-18 nucleotide units in length.

30. The gapmer oligonucleotide according to claim 11 , wherein the oligomer is 13, 14, 15, 16 or 17 nucleotide units in length.

Assignees

• Roche Innovation Ct Copenhagen As

Inventors

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

Classifications

• C07H21/00

  Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids · CPC title

• C12N15/113 Primary

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• C12N15/1137

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

• A61K31/7088

  Compounds having three or more nucleosides or nucleotides · CPC title

• C12N2320/51

  modulating the chemical stability, e.g. nuclease-resistance · CPC title