Antisense molecules and methods for treating pathologies

• US 8,637,483 B2
• Filed: 11/12/2010
• Issued: 01/28/2014
• Est. Priority Date: 11/12/2009
• Status: Active Grant

• Current Assignee: University of Western Australia
• Original Assignee: University of Western Australia
• Inventors: Wilton, Stephen Donald, Fletcher, Sue, Adams, Abbie, Meloni, Penny
• Primary Examiner(s): VIVLEMORE, TRACY ANN
• Application Number:
  US13/509,331
• Publication Number:
  US 20120270925A1
• Time in Patent Office: 1,173 Days
• Field of Search: None
• US Class Current: 514/44.A
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• 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 2310/3181 Peptide nucleic acid, PNA
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/351 Conjugate
• CPC Class Codes: C12N 2310/3521 Methyl
• 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 59.

Claims

1. A combination of two or more antisense molecules selected from the following combinations:

(a) an antisense oligonucleotide comprising SEQ ID NO;

31 and an antisense oligonucleotide comprising SEQ ID NO;

32;

(b) an antisense oligonucleotide comprising SEQ ID NO;

33 and an antisense oligonucleotide comprising SEQ ID NO;

34;

(c) an antisense oligonucleotide comprising SEQ ID NO;

35 and an antisense oligonucleotide comprising SEQ ID NO;

36;

(d) an antisense oligonucleotide comprising SEQ ID NO;

39, an antisense oligonucleotide comprising SEQ ID NO;

40, and an antisense oligonucleotide comprising SEQ ID NO;

41;

(e) an antisense oligonucleotide comprising SEQ ID NO;

42 and an antisense oligonucleotide comprising SEQ ID NO;

43;

(f) an antisense oligonucleotide comprising SEQ ID NO;

44 and an antisense oligonucleotide comprising SEQ ID NO;

45;

(g) an antisense oligonucleotide comprising SEQ ID NO;

46 and an antisense oligonucleotide comprising SEQ ID NO;

47;

(h) an antisense oligonucleotide comprising SEQ ID NO;

48 and an antisense oligonucleotide comprising SEQ ID NO;

49; and

(i) an antisense oligonucleotide comprising SEQ ID NO;

50 and an antisense oligonucleotide comprising SEQ ID NO;

51,

wherein each of the antisense oligonucleotides comprises a modification to minimize or prevent cleavage by RNase H, and wherein the combination of antisense molecules is capable of binding to selected targets in dystrophin pre-mRNA to induce exon skipping in the human dystrophin gene.

2. The antisense molecules according to claim 1, capable of binding to selected target sites, wherein the target site is an mRNA splicing element selected from a splice donor site, splice acceptor sites or exonic splicing enhancer elements.

3. A pharmaceutical composition for the treatment of muscular dystrophy in a patient comprising (a) a combination of antisense molecules according to claim 1, and (b) one or more pharmaceutically acceptable carriers and/or diluents.

4. A method of treating muscular dystrophy in a patient comprising administering to the patient a pharmaceutical composition according to claim 3.

5. A kit comprising a combination of antisense molecules according to claim 1, a suitable carrier and instructions for it'"'"'s use.

6. The combination of claim 1, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

31 and an antisense oligonucleotide comprising SEQ ID NO;

32, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

7. The combination of claim 1, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

33 and an antisense oligonucleotide comprising SEQ ID NO;

34, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

8. The combination of claim 1, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

35 and an antisense oligonucleotide comprising SEQ ID NO;

36, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

9. The combination of claim 1, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

39, an antisense oligonucleotide comprising SEQ ID NO;

40, and an antisense oligonucleotide comprising SEQ ID NO;

41, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

10. The combination of claim 1, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

42 and an antisense oligonucleotide comprising SEQ ID NO;

43, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

11. The combination of claim 1, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

44 and an antisense oligonucleotide comprising SEQ ID NO;

45, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

12. The combination of claim 1, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

46 and an antisense oligonucleotide comprising SEQ ID NO;

47, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

13. The combination of claim 1, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

48 and an antisense oligonucleotide comprising SEQ ID NO;

49, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

14. The combination of claim 1, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

50 and an antisense oligonucleotide comprising SEQ ID NO;

51, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

15. The combination of claim 1, wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

16. The combination of claim 1, wherein the antisense oligonucleotides comprise a nucleobase modification or nucleobase substitution.

17. The combination of claim 1, wherein the antisense oligonucleotides comprise a modified backbone.

18. The combination of claim 1, wherein the antisense oligonucleotides comprise non-natural inter-nucleoside linkages.

19. The combination of claim 1, wherein the antisense oligonucleotides comprise modified backbones in which the inter-nucleotide bridging phosphate residues are modified phosphates.

20. The combination of claim 19, wherein the modified phosphates are methyl phosphonates, methyl phosphorothioates, phosphoromorpholidates, phosphoropiperazidates or phosphoroamidates.

21. The combination of claim 1, wherein the antisense oligonucleotides are 2′

-O-methyl-oligoribonucleotides.

22. The combination of claim 1, wherein the antisense oligonucleotides are peptide nucleic acids.

23. The combination of claim 1, wherein the antisense oligonucleotides are chemically linked to one or more moieties or conjugates that enhance the activity, cellular distribution, or cellular uptake of the antisense oligonucleotide.

24. The combination of claim 1, wherein the antisense oligonucleotides are conjugated to a polyamine.

25. The combination of claim 1, wherein the antisense oligonucleotides are chemically linked to a polyethylene glycol chain.

26. A pharmaceutical composition comprising a combination of antisense molecules according to claim 6, and a pharmaceutically acceptable carrier.

27. A pharmaceutical composition comprising a combination of antisense molecules according to claim 7, and a pharmaceutically acceptable carrier.

28. A pharmaceutical composition comprising a combination of antisense molecules according to claim 8, and a pharmaceutically acceptable carrier.

29. A pharmaceutical composition comprising a combination of antisense molecules according to claim 9, and a pharmaceutically acceptable carrier.

30. A pharmaceutical composition comprising a combination of antisense molecules according to claim 10, and a pharmaceutically acceptable carrier.

31. A pharmaceutical composition comprising a combination of antisense molecules according to claim 11, and a pharmaceutically acceptable carrier.

32. A pharmaceutical composition comprising a combination of antisense molecules according to claim 12, and a pharmaceutically acceptable carrier.

33. A pharmaceutical composition comprising a combination of antisense molecules according to claim 13, and a pharmaceutically acceptable carrier.

34. A pharmaceutical composition comprising a combination of antisense molecules according to claim 14, and a pharmaceutically acceptable carrier.

35. The method of claim 4, wherein the muscular dystrophy is Duchenne muscular dystrophy.

36. A method of treating muscular dystrophy in a patient comprising administering an effective amount of a combination of two or more antisense molecules selected from the following combinations:

(a) an antisense oligonucleotide comprising SEQ ID NO;

31 and an antisense oligonucleotide comprising SEQ ID NO;

32;

(b) an antisense oligonucleotide comprising SEQ ID NO;

33 and an antisense oligonucleotide comprising SEQ ID NO;

34;

(c) an antisense oligonucleotide comprising SEQ ID NO;

35 and an antisense oligonucleotide comprising SEQ ID NO;

36;

(d) an antisense oligonucleotide comprising SEQ ID NO;

39, an antisense oligonucleotide comprising SEQ ID NO;

40 and an antisense oligonucleotide comprising SEQ ID NO;

41;

(e) an antisense oligonucleotide comprising SEQ ID NO;

42 and an antisense oligonucleotide comprising SEQ ID NO;

43;

(f) an antisense oligonucleotide comprising SEQ ID NO;

44 and an antisense oligonucleotide comprising SEQ ID NO;

45;

(g) an antisense oligonucleotide comprising SEQ ID NO;

46 and an antisense oligonucleotide comprising SEQ ID NO;

47;

(h) an antisense oligonucleotide comprising SEQ ID NO;

48 and an antisense oligonucleotide comprising SEQ ID NO;

49; and

(i) an antisense oligonucleotide comprising SEQ ID NO;

50 and an antisense oligonucleotide comprising SEQ ID NO;

51,

wherein each of the antisense oligonucleotides comprises a modification to minimize or prevent cleavage by RNase H, and wherein the combination of antisense molecules is capable of binding to a selected target in dystrophin pre-mRNA to induce exon skipping in the human dystrophin gene.

37. The method of claim 36, wherein the muscular dystrophy is Duchenne muscular dystrophy.

38. The method of claim 36, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

31 and an antisense oligonucleotide comprising SEQ ID NO;

32, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

39. The method of claim 36, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

33 and an antisense oligonucleotide comprising SEQ ID NO;

34, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

40. The method of claim 36, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

35 and an antisense oligonucleotide comprising SEQ ID NO;

36, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

41. The method of claim 36, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

39, an antisense oligonucleotide comprising SEQ ID NO;

40, and an antisense oligonucleotide comprising SEQ ID NO;

41, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

42. The method of claim 36, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

42 and an antisense oligonucleotide comprising SEQ ID NO;

43, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

43. The method of claim 36, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

44 and an antisense oligonucleotide comprising SEQ ID NO;

45, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

44. The method of claim 36, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

46 and an antisense oligonucleotide comprising SEQ ID NO;

47, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

45. The method of claim 36, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

48 and an antisense oligonucleotide comprising SEQ ID NO;

49, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.

46. The method of claim 36, wherein the antisense molecules are an antisense oligonucleotide comprising SEQ ID NO:

50 and an antisense oligonucleotide comprising SEQ ID NO;

51, and wherein the antisense oligonucleotides comprise deoxyribonucleotide or ribonucleotide sequences.