MULTIPLE EXON SKIPPING COMPOSITIONS FOR DMD

• US 20140094500A1
• Filed: 03/14/2013
• Published: 04/03/2014
• Est. Priority Date: 10/24/2008
• Status: Abandoned Application

• Current Assignee: Sarepta Therapeutics, Inc.
• Original Assignee: Sarepta Therapeutics, Inc.
• Inventors: SAZANI, Peter, KOLE, Ryszard
• Application Number:
  US13/829,545
• Publication Number:
  US 20140094500A1
• Time in Patent Office: Days
• 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 2310/321 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. An isolated antisense compound of 20 to 35 nucleotides in length comprising at least 12 contiguous nucleotides of a nucleotide sequence selected from the group consisting of SEQ ID NOs:

1-569 and 612-635, wherein the oligonucleotide specifically hybridizes to a target region in an exon of the human dystrophin gene inducing exon skipping.

2. The antisense compound of claim 1, wherein the sequence is selected from the group consisting of SEQ ID NOs:

1-20, 21-76, 612-624, 77-125, 126-169, 170-224, 634, 225-266, 267-308, 309-371, 372-415, 416-475, 625-633, 476-519, 520-569, and 635.

3. The antisense compound of claim 1, wherein the exon in human dystrophin is selected from the group consisting of exons 44-55.

4. The antisense compound of claim 1, which is substantially uncharged.

5. The antisense compound of claim 4, which comprises morpholino subunits linked by phosphorus-containing intersubunit linkages joining a morpholino nitrogen of one subunit to a 5′

exocyclic carbon of an adjacent subunit.

6. The antisense compound of claim 4, which comprises morpholino subunits linked by substantially uncharged phosphorus-containing intersubunit linkages joining a morpholino nitrogen of one subunit to a 5′

exocyclic carbon of an adjacent subunit.

7. The antisense compound of claim 6, wherein 3-35% of the linkages are positively charged.

8. The antisense compound of claim 5, 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.

9. The antisense compound of claim 6, 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.

10. The antisense compound of claim 1, comprising morpholino subunits and phosphorodiamidate intersubunit linkages.

11. The antisense compound of claim 5, wherein at least one and up to 50% of the intersubunit linkages are modified with a pendent cationic group.

12. The antisense compound of claim 6, wherein at least one and up to 50% of the intersubunit linkages are modified with a pendent cationic group.

13. The antisense compound of claim 1, which is chemically linked to one or more moieties or conjugates that enhance the activity, cellular distribution, or cellular uptake of the antisense oligonucleotide.

14. The antisense compound of claim 13, which is conjugated to a polyamine.

15. The antisense compound of claim 13, which is chemically linked to polyethylene glycol.

18. The antisense compound of claim 1 which specifically hybridizes to a target region in exon 55 and the sequence is selected from the group consisting of SEQ ID NO:

520-569 and 635.

19. A pharmaceutical composition, comprising an antisense compound of claim 1, and a saline solution that includes a phosphate buffer.

20. A method of treating Duchenne muscular dystrophy, comprising administering to a patient in need thereof an effective amount of a pharmaceutical composition of claim 19.

16. The antisense compound of claim 27, which is conjugated to an arginine-rich peptide.