Multiple exon skipping compositions for DMD
View Patent ↗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.
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).
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.
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.
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.
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).