Antisense oligonucleotides for inducing exon skipping and methods of use thereof

• US 9,175,286 B2
• Filed: 03/14/2013
• Issued: 11/03/2015
• 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, Fletcher, Sue, McClorey, Graham
• Primary Examiner(s): Chong, Kimberly
• Application Number:
  US13/826,880
• Publication Number:
  US 20130274313A1
• Time in Patent Office: 964 Days
• US Class Current: 1/1
• 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. An antisense oligonucleotide of 24 bases comprising the base sequence GAU AGG UGG UAU CAA CAU CUG UAA (SEQ ID NO:

1), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide is a morpholino antisense oligonucleotide.

2. The antisense oligonucleotide of claim 1, 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.

3. The antisense oligonucleotide of claim 1, wherein the oligonucleotide is chemically linked to a polyethylene glycol chain.

4. An antisense oligonucleotide of 21 bases comprising the base sequence GAU AGG UGG UAU CAA CAU CUG (SEQ ID NO:

2), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide is a morpholino antisense oligonucleotide.

5. The antisense oligonucleotide of claim 4, 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.

6. The antisense oligonucleotide of claim 4, wherein the oligonucleotide is chemically linked to a polyethylene glycol chain.

7. An antisense oligonucleotide of 25 bases comprising the base sequence GAU AGG UGG UAU CAA CAU CUG UAA G (SEQ ID NO:

3), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide is a morpholino antisense oligonucleotide.

8. The antisense oligonucleotide of claim 7, 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.

9. The antisense oligonucleotide of claim 7, wherein the oligonucleotide is chemically linked to a polyethylene glycol chain.

10. An antisense oligonucleotide of 20 bases comprising the base sequence GGU GGU AUC AAC AUC UGU AA (SEQ ID NO:

4), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide is a morpholino antisense oligonucleotide.

11. The antisense oligonucleotide of claim 10, 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.

12. The antisense oligonucleotide of claim 10, wherein the oligonucleotide is chemically linked to a polyethylene glycol chain.

13. A pharmaceutical composition comprising an antisense oligonucleotide of 20 to 50 bases comprising at least 17 consecutive bases of the base sequence GAU AGG UGG UAU CAA CAU CUG UAA (SEQ ID NO:

1), in which the uracil bases are optionally thymine bases, wherein the antisense oligonucleotide is a morpholino antisense oligonucleotide, and wherein the antisense oligonucleotide specifically hybridizes to a target region in exon 8 of the human dystrophin gene inducing exon 8 skipping, and a pharmaceutically acceptable carrier.

14. The pharmaceutical composition of claim 13, 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.

15. The pharmaceutical composition of claim 13, wherein the oligonucleotide is chemically linked to a polyethylene glycol chain.

16. A pharmaceutical composition comprising an antisense oligonucleotide of 21 bases comprising the base sequence AGG UGG UAU CAA CAU CUG (SEQ ID NO:

2), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide is a morpholino antisense oligonucleotide, and a pharmaceutically acceptable carrier.

17. The pharmaceutical composition of claim 16, 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 pharmaceutical composition of claim 16, wherein the oligonucleotide is chemically linked to a polyethylene glycol chain.

19. A pharmaceutical composition comprising an antisense oligonucleotide of 25 bases comprising the base sequence GAU AGG UGG UAU CAA CAU CUG UAA G (SEQ ID NO:

3), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide is a morpholino antisense oligonucleotide, and a pharmaceutically acceptable carrier.

20. The pharmaceutical composition of claim 19, 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.

21. The pharmaceutical composition of claim 19, wherein the oligonucleotide is chemically linked to a polyethylene glycol chain.

22. A pharmaceutical composition comprising an antisense oligonucleotide of 20 bases comprising the base sequence GGU GGU AUC AAC AUC UGU AA (SEQ ID NO:

4), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide is a morpholino antisense oligonucleotide, and a pharmaceutically acceptable carrier.

23. The pharmaceutical composition of claim 22, 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.

24. The pharmaceutical composition of claim 22, wherein the oligonucleotide is chemically linked to a polyethylene glycol chain.