EXON SKIPPING COMPOSITIONS FOR TREATING MUSCULAR DYSTROPHY

• US 20160040162A1
• Filed: 03/14/2014
• Published: 02/11/2016
• Est. Priority Date: 03/14/2013
• Status: Abandoned Application

• Current Assignee: Sarepta Therapeutics, Inc.
• Original Assignee: Sarepta Therapeutics, Inc.
• Inventors: BESTWICK, Richard K., FRANK, Diane Elizabeth
• Application Number:
  US14/776,533
• Publication Number:
  US 20160040162A1
• Time in Patent Office: Days
• US Class Current: 1/1
• 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 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/351 Conjugate
• CPC Class Codes: C12N 2310/3513 Protein; Peptide
• CPC Class Codes: C12N 2310/3535 Nitrogen
• CPC Class Codes: C12N 2320/33 Alteration of splicing

Abstract

Antisense molecules capable of binding to a selected target site in the human dystrophin gene to induce exon 53 skipping are described.

Claims

1. An antisense oligomer of 20-50 nucleotides in length capable of binding a selected target to induce exon skipping in the human dystrophin gene, wherein said antisense oligomer comprises a sequence of bases that specifically hybridizes to an exon 53 target region selected from the group consisting of H53A(+36+60), H53A(+30+57), H53A (+30+56), H53A(+30+55) and H53A(+33+57), wherein the bases of said oligomer are linked to morpholino ring structures, and wherein said morpholino ring structures are joined by phosphorous-containing intersubunit linkages joining a morpholino nitrogen of one ring structure to a 5′

exocyclic carbon of an adjacent ring structure.

2. The antisense oligomer of claim 1, wherein the sequence is SEQ ID NO:

1-5.

3. The antisense oligomer of claim 1 or 2, which is about 20 to 30 nucleotides in length.

4. The antisense oligomer of claim 1 or 2, which is about 25 to 28 nucleotides in length.

5. The antisense oligomer of claim 2, wherein the sequence consists of a sequence selected from SEQ ID NO:

1-5.

6. The antisense oligomer of any one of claims 1-5, wherein the oligomer does not activate RNase H.

7. The antisense oligomer of any one of claims 1-5, wherein the oligomer is chemically linked to one or more moieties or conjugates that enhance the activity, cellular distribution, or cellular uptake of the antisense oligomer.

8. The antisense oligomer of claim 7, wherein the oligomer is chemically linked to a polyethylene glycol molecule.

9. The antisense oligomer of any one of claims 1-5, wherein the antisense oligomer is conjugated to an arginine-rich peptide.

10. The antisense oligomer of claim 9, wherein the arginine-rich peptide comprises a sequence selected from SEQ ID NOS:

16-31.

11. The antisense oligomer of any of the preceding claims, comprising morpholino ring structures joined by substantially uncharged phosphorus-containing intersubunit linkages joining a morpholino nitrogen of one ring structure to a 5′

exocyclic carbon of an adjacent ring structure.

12. The antisense oligomer of claim 11, wherein 5%-35% of the linkages is positively charged.

13. The antisense oligomer of any of the preceding claims, wherein the intersubunit linkages are uncharged and interspersed with linkages that are positively charged at physiological pH, wherein the total number of positively charged linkages is between 2 and no more than half of the total number of linkages.

14. The antisense oligomer of any one of claims 1-13, comprising morpholino ring structures and phosphorodiamidate intersubunit linkages.

17. A composition comprising the antisense oligomer of any of the preceding claims and a pharmaceutically acceptable carrier.

18. The composition of claim 17 for use in the treatment of muscular dystrophy.

19. The composition of claim 18, wherein the muscular dystrophy is Duchenne'"'"'s muscular dystrophy (DMD).

20. The composition of claim 18, wherein the muscular dystrophy is Becker'"'"'s muscular dystrophy (BMD).

15. The antisense oligomer are modified with a pendant cationic group.

16. An antisense oligomer, wherein the oligomer has a structure selected from the group consisting of: