ANTISENSE OLIGONUCLEOTIDES FOR INDUCING EXON SKIPPING AND METHODS OF USE THEREOF

• US 20110046203A1
• Filed: 08/20/2010
• Published: 02/24/2011
• Est. Priority Date: 06/28/2004
• Status: Active Grant

• Current Assignee: University of Western Australia
• Original Assignee: University of Western Australia
• Inventors: Wilton, Stephen Donald, McClorey, Graham, Fletcher, Sue
• Granted Patent: US 7,960,541 B2
• Time in Patent Office: Days
• US Class Current: 514/44.00A
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: C12N 15/113 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 Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/33 of the base
• CPC Class Codes: C12N 2310/3341 5-Methylcytosine
• CPC Class Codes: C12N 2310/3519 Fusion with another nucleic acid
• CPC Class Codes: C12N 2320/30 Special therapeutic applications
• CPC Class Codes: C12N 2320/33 Alteration of splicing

Abstract

An antisense molecule capable of binding to a selected target site to induce exon skipping in the dystrophin gene, as set forth in SEQ ID NO: 1 to 202.

Claims

1-14. -14. (canceled)

15. An isolated antisense oligonucleotide of 25 to 50 nucleotides in length comprising SEQ ID NO:

180, wherein the uracil bases are optionally thymine bases.

16. The antisense oligonucleotide of claim 15, wherein the antisense oligonucleotide comprises a non-natural backbone.

17. The antisense oligonucleotide of claim 15, wherein the antisense oligonucleotide is chemically linked to one or more moieties or conjugates that enhance the activity, cellular distribution, or cellular uptake of the antisense oligonucleotide.

18. The antisense oligonucleotide of claim 15, wherein the antisense oligonucleotide does not activate RNase H.

19. The antisense oligonucleotide of claim 16, wherein the sugar moieties of the oligonucleotide backbone are replaced with non-natural moieties.

20. The antisense oligonucleotide of claim 19, wherein the non-natural moieties are morpholinos.

21. The antisense oligonucleotide of claim 20, wherein the uracil bases are thymine bases.

22. The antisense oligonucleotide of claim 15, wherein the inter-nucleotide linkages of the oligonucleotide backbone are replaced with non-natural inter-nucleotide linkages.

23. The antisense oligonucleotide of claim 22, wherein the non-natural inter-nucleotide linkages are modified phosphates.

24. The antisense oligonucleotide of claim 23, wherein the modified phosphates are methyl phosphonates, methyl phosphorothioates, phosphoromorpholidates, phosphoropiperazidates or phosphoroamidates.

25. The antisense oligonucleotide of claim 24, wherein the modified phosphates are phosphoroamidates.

26. The antisense oligonucleotide of claim 24, wherein the modified phosphates are phosphoromorpholidates.

27. The antisense oligonucleotide of claim 15, wherein the sugar moieties of the oligonucleotide backbone are replaced with non-natural moieties and the inter-nucleotide linkages of the oligonucleotide backbone are replaced with non-natural inter-nucleotide linkages.

28. The antisense oligonucleotide of claim 27, wherein the non-natural moieties are morpholinos and the non-natural internucleotide linkages are modified phosphates.

29. The antisense oligonucleotide of claim 28, wherein the modified phosphates are methyl phosphonates, methyl phosphorothioates, phosphoromorpholidates, phosphoropiperazidates or phosphoroamidates.

30. The antisense oligonucleotide of claim 29, wherein the modified phosphates are phosphoroamidates.

31. The antisense oligonucleotide of claim 29, wherein the modified phosphates are phosphoromorpholidates.

32. The antisense oligonucleotide of claim 29, wherein the uracil bases are thymine bases.

33. The antisense oligonucleotide of claim 32, wherein the antisense oligonucleotide is chemically linked to one or more moieties or conjugates that enhance the activity, cellular distribution, or cellular uptake of the antisense oligonucleotide.

34. The antisense oligonucleotide of claim 15, wherein the antisense oligonucleotide is 25 nucleotides in length.

35. The antisense oligonucleotide of claim 34, wherein the uracil bases are thymine bases.

36. A pharmaceutical composition, comprising the antisense oligonucleotide of claim 15 and a pharmaceutically acceptable carrier.

37. The pharmaceutical composition of claim 36, wherein the pharmaceutically acceptable carrier is phosphate-buffered saline.

38. A pharmaceutically acceptable salt of the antisense oligonucleotide of claim 15.