Multiple exon skipping compositions for DMD

• US 8,871,918 B2
• Filed: 10/23/2009
• Issued: 10/28/2014
• Est. Priority Date: 10/24/2008
• Status: Active Grant

• Current Assignee: Sarepta Therapeutics, Inc.
• Original Assignee: Sarepta Therapeutics, Inc.
• Inventors: Sazani, Peter, Kole, Ryszard
• Primary Examiner(s): MCDONALD, JENNIFER SUE PITRAK
• Application Number:
  US12/605,276
• Publication Number:
  US 20100130591A1
• Time in Patent Office: 1,831 Days
• US Class Current: 536/23.1
• 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 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/331 Universal or degenerate base
• CPC Class Codes: C12N 2310/3341 5-Methylcytosine
• CPC Class Codes: C12N 2310/3513 Protein; Peptide
• CPC Class Codes: C12N 2320/30 Special therapeutic applications
• CPC Class Codes: C12N 2320/33 Alteration of splicing

Abstract

Provided are antisense molecules capable of binding to a selected target site in the human dystrophin gene to induce exon skipping, and methods of use thereof to treat muscular dystrophy.

Claims

1. A composition for producing skipping of exon 50 in the processing of human dystrophin pre-processed mRNA, comprising a pharmaceutically acceptable carrier and

a substantially uncharged antisense compound consisting of a base sequence set forth in SEQ ID NO:

287 and consisting of 25 morpholino subunits linked by phosphorus-containing intersubunit linkages joining a morpholino nitrogen of one subunit to a 5′

exocyclic carbon of an adjacent subunit, wherein

the antisense compound is capable of forming a heteroduplex structure with a complementary mRNA sequence in the dystrophin-gene exon 50.

2. The composition of claim 1, wherein the antisense compound is conjugated to an arginine-rich peptide.

3. The composition of claim 2, wherein the arginine-rich peptide comprises a sequence selected from SEQ ID NOS:

570-578.

4. The composition of claim 1, wherein the intersubunit linkages are phosphorodiamidate intersubunit linkages.

5. The composition of claim 4, wherein at least one and up to about 50% of the intersubunit linkage(s) comprise a pendant cationic group.

6. The composition of claim 5, wherein the cationic group is 1-piperazinyl.

7. A method of inducing exon 50 skipping in a subject with muscular dystrophy, comprising administering to the subject an effective amount of a composition according to claim 1.

8. The method of claim 7, wherein the muscular dystrophy is Duchenne'"'"'s muscular dystrophy (DMD).

9. The method of claim 7, wherein the muscular dystrophy is Becker muscular dystrophy (BMD).

16. The composition of claim 1, wherein the antisense compound is conjugated to a chemical moiety.

17. The composition of claim 16, wherein the chemical moiety is a polyethylene glycol moiety.

18. The composition of claim 4, wherein the antisense compound is conjugated to a chemical moiety.

19. The composition of claim 18, wherein the chemical moiety is a polyethylene glycol moiety.

24. A method of inducing exon 50 skipping in a subject with muscular dystrophy, comprising administering to the subject an effective amount of a composition according to claim 4.

25. The method of claim 24, wherein the muscular dystrophy is Duchenne'"'"'s muscular dystrophy (DMD).

26. The method of claim 24, wherein the muscular dystrophy is Becker muscular dystrophy (BMD).

27. A method of inducing exon 50 skipping in a subject with muscular dystrophy, comprising administering to the subject an effective amount of a composition according to claim 16.

28. The method of claim 27, wherein the muscular dystrophy is Duchenne'"'"'s muscular dystrophy (DMD).

29. The method of claim 27, wherein the muscular dystrophy is Becker muscular dystrophy (BMD).

30. A method of inducing exon 50 skipping in a subject with muscular dystrophy, comprising administering to the subject an effective amount of a composition according to claim 17.

31. The method of claim 30, wherein the muscular dystrophy is Duchenne'"'"'s muscular dystrophy (DMD).

32. The method of claim 30, wherein the muscular dystrophy is Becker muscular dystrophy (BMD).

33. A method of inducing exon 50 skipping in a subject with muscular dystrophy, comprising administering to the subject an effective amount of a composition according to claim 18.

34. The method of claim 33, wherein the muscular dystrophy is Duchenne'"'"'s muscular dystrophy (DMD).

35. The method of claim 33, wherein the muscular dystrophy is Becker muscular dystrophy (BMD).

36. A method of inducing exon 50 skipping in a subject with muscular dystrophy, comprising administering to the subject an effective amount of a composition according to claim 19.

37. The method of claim 36, wherein the muscular dystrophy is Duchenne'"'"'s muscular dystrophy (DMD).

38. The method of claim 36, wherein the muscular dystrophy is Becker muscular dystrophy (BMD).

10. A substantially uncharged antisense oligonucleotide consisting of a base sequence set forth in SEQ ID NO:

287 and consisting of 25 morpholino subunits linked by phosphorus-containing intersubunit linkages joining a morpholino nitrogen of one subunit to a 5′

exocyclic carbon of an adjacent subunit, wherein the antisense oligonucleotide is capable of forming a heteroduplex structure with a complementary mRNA sequence in the dystrophin-gene exon 50 inducing exon skipping.

11. The antisense oligonucleotide of claim 10, wherein the oligonucleotide is conjugated to an arginine-rich peptide.

12. The antisense oligonucleotide of claim 11, wherein the arginine-rich peptide comprises a sequence selected from SEQ ID NOS:

570-578.

13. The antisense oligonucleotide of claim 10, wherein the intersubunit linkages are phosphorodiamidate intersubunit linkages.

14. The antisense oligonucleotide of claim 13, wherein at least one and up to about 50% of the intersubunit linkage(s) comprise a pendant cationic group.

15. The antisense oligonucleotide of claim 14, wherein the cationic group is 1-piperazinyl.

20. The antisense oligonucleotide of claim 10, wherein the oligonucleotide is conjugated to a chemical moiety.

21. The antisense oligonucleotide of claim 20, wherein the chemical moiety is a polyethylene glycol moiety.

22. The antisense oligonucleotide of claim 13, wherein the oligonucleotide is conjugated to a chemical moiety.

23. The antisense oligonucleotide of claim 22, wherein the chemical moiety is a polyethylene glycol moiety.