Interfering RNA molecules

Patent metadata

Field	Value
Publication number	US-9695423-B2
Application number	US-201514977710-A
Country	US
Kind code	B2
Filing date	Dec 22, 2015
Priority date	Aug 5, 2002
Publication date	Jul 4, 2017
Grant date	Jul 4, 2017

Abstract

The present invention is related to a ribonucleic acid comprising a double stranded structure whereby the double-stranded structure comprises a first strand and a second strand, whereby the first strand comprises a first stretch of contiguous nucleotides and whereby said first stretch is at least partially complementary to a target nucleic acid, and the second strand comprises a second stretch of contiguous nucleotides whereby said second stretch is at least partially identical to a target nucleic acid, and whereby the double stranded structure is blunt ended.

First claim

We claim:

1. A double-stranded siRNA molecule wherein:
(i) at least one strand of the double-stranded siRNA molecule comprises one or more groups of modified nucleotides and one or more groups of flanking nucleotides, the flanking nucleotides being on one or both sides of the modified nucleotides, wherein:
the one or more groups of modified nucleotides have an amino, fluoro, alkoxy, or alkyl modification at the 2′-position; and
the one or more groups of flanking nucleotides have an amino, fluoro, alkoxy, or alkyl modification at the 2′-position, the modification at the 2′-position of the flanking nucleotide being different from the modification at the 2′-position of the modified nucleotide,
(ii) the number of nucleotides in the one or more groups of modified nucleotides is 1-10 and the number of nucleotides in the one or more groups of flanking nucleotides is 1-10;
(iii) the double-stranded siRNA molecule has a double-stranded region of 17-21 nucleotides in length; and
(iv) the double-stranded siRNA molecule is capable of RNA interference.

2. The double-stranded siRNA molecule of claim 1 , wherein the double-stranded region is at least 18 nucleotides in length.

3. The double-stranded siRNA molecule of claim 2 , wherein the alkoxy modification at the 2′-position of the one or more groups of modified nucleotides or the one or more groups of flanking nucleotides is a methoxy modification.

4. The double-stranded siRNA molecule of claim 2 , wherein the at least one strand is the antisense strand.

5. The double-stranded siRNA molecule of claim 2 , wherein at least one of the strands has an overhang of at least one nucleotide at the 3′-end.

6. The double-stranded siRNA molecule of claim 5 , wherein the strand having the overhang at the 3′-end is the antisense strand.

7. The double-stranded siRNA molecule of claim 2 , wherein the one or more groups of modified nucleotides have an alkoxy modification at the 2′-position, the alkoxy modification being a methoxy modification.

8. The double-stranded siRNA molecule of claim 7 , wherein the one or more groups of flanking nucleotides have a fluoro modification at the 2′-position.

9. The double-stranded siRNA molecule of claim 2 , wherein the one or more groups of modified nucleotides are located on at least alternating nucleotide positions over the length of the at least one strand.

10. The double-stranded siRNA molecule of claim 2 , wherein the double-stranded siRNA molecule has increased stability in serum and/or cell culture medium compared to a double-stranded siRNA molecule lacking the one or more groups of modified nucleotides and/or the one or more groups of flanking nucleotides.

11. The double-stranded siRNA molecule of claim 2 , wherein the double-stranded siRNA molecule inhibits the expression of a target nucleic acid.

12. A double-stranded siRNA molecule wherein:
(i) each strand of the double-stranded siRNA molecule comprises one or more groups of modified nucleotides and one or more groups of flanking nucleotides, the flanking nucleotides being on one or both sides of the modified nucleotides, wherein
the one or more groups of modified nucleotides have an amino, fluoro, alkoxy, or alkyl modification at the 2′-position; and
the one or more groups of flanking nucleotides have an amino, fluoro, alkoxy, or alkyl modification at the 2′-position, the modification at the 2′-position of the flanking nucleotide being different from the modification at the 2′-position of the modified nucleotide,
(ii) the number of nucleotides in the one or more groups of modified nucleotides is 1-10 and the number of nucleotides in the one or more groups of flanking nucleotides is 1-10;
(iii) the double-stranded siRNA molecule has a double-stranded region of 17-21 nucleotides in length; and
(iv) the double-stranded siRNA molecule is capable of RNA interference.

13. The double-stranded siRNA molecule of claim 12 , wherein the double-stranded region is at least 18 nucleotides in length.

14. The double-stranded siRNA molecule of claim 13 , wherein the alkoxy modification at the 2′-position of the one or more groups of modified nucleotides or the one or more groups of flanking nucleotides is a methoxy modification.

15. The double-stranded siRNA molecule of claim 13 , wherein at least one of the strands has an overhang of at least one nucleotide at the 3′-end.

16. The double-stranded siRNA molecule of claim 15 , wherein the strand having the overhang at the 3′-end is the antisense strand.

17. The double-stranded siRNA molecule of claim 13 , wherein the one or more groups of modified nucleotides on at least one of the strands have an alkoxy modification at the 2′-position, the alkoxy modification being a methoxy modification.

18. The double-stranded siRNA molecule of claim 13 , wherein, on at least one of the strands, the one or more groups of modified nucleotides have a methoxy modification at the 2′-position and the one or more groups of flanking nucleotides have a fluoro modification at the 2′-position.

19. The double-stranded siRNA molecule of claim 13 , wherein the one or more groups of modified nucleotides on at least one of the strands are located on at least alternating nucleotide positions over the length of the strand.

20. The double-stranded siRNA molecule of claim 13 , wherein the double-stranded siRNA molecule has increased stability in serum and/or cell culture medium compared to a double-stranded siRNA molecule lacking the one or more groups of modified nucleotides and/or the one or more groups of flanking nucleotides.

21. The double-stranded siRNA molecule of claim 13 , wherein the double-stranded siRNA molecule inhibits the expression of a target nucleic acid.

22. The double-stranded siRNA molecule of claim 1 , wherein the double-stranded siRNA molecule inhibits the expression of a target nucleic acid.

23. The double-stranded siRNA molecule of claim 12 , wherein the double-stranded siRNA molecule inhibits the expression of a target nucleic acid.

24. The double-stranded siRNA molecule of claim 1 , wherein the one or more groups of modified nucleotides have a methoxy modification at the 2′-position and the one or more groups of flanking nucleotides have a fluoro modification at the 2′-position.

25. The double-stranded siRNA molecule of claim 12 , wherein, on at least one of the strands, the one or more groups of modified nucleotides have a methoxy modification at the 2′-position and the one or more groups of flanking nucleotides have a fluoro modification at the 2′-position.

Assignees

• Silence Therapeutics Gmbh

Inventors

• Giese Klaus
• Kaufmann Jörg
• Klippel-Giese Anke

Classifications

• A61P35/02

  specific for leukemia · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P3/04

  Anorexiants; Antiobesity agents · CPC title