Claims for Patent: RE48960

Summary for Patent: RE48960

Title:	Antisense oligonucleotides for inducing exon skipping and methods of use thereof
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 202.
Inventor(s):	Wilton; Stephen Donald (Applecross, AU), Fletcher; Sue (Bayswater, AU), McClorey; Graham (Bayswater, AU)
Assignee:	The University of Western Australia (Crawley, AU)
Application Number:	16/721,681
Patent Claims:	1. An isolated antisense oligonucleotide of 20 to .[.50 nucleotides in length comprising.]. .Iadd.31 bases comprising a base sequence that is 100% complementary to consecutive bases of exon 45 of the human dystrophin pre-mRNA, wherein the base sequence comprises .Iaddend.at least .[.17.]. .Iadd.20 .Iaddend.consecutive .[.nucleotides of.]. .Iadd.bases of CCA AUG CCA UCC UGG AGU UCC UGU AA (.Iaddend.SEQ ID NO: 207.Iadd.).Iaddend., .Iadd.in which uracil bases are thymine bases, wherein the antisense oligonucleotide is a morpholino antisense oligonucleotide, and .Iaddend.wherein the .Iadd.antisense .Iaddend.oligonucleotide .[.specifically hybridizes to an exon 45 acceptor splice site of a human dystrophin gene, inducing.]. .Iadd.induces .Iaddend.exon 45 skipping.[., and wherein the uracil bases are optionally thymine bases.].. .[.2. The antisense oligonucleotide of claim 1, wherein the uracil bases are thymine bases..]. .[.3. The antisense oligonucleotide of claim 1 comprising SEQ ID NO: 207..]. .[.4. The antisense oligonucleotide of claim 1 consisting of SEQ ID NO: 207..]. .[.5. The antisense oligonucleotide of claim 1 comprising SEQ ID NO: 207, wherein the uracil bases are thymine bases..]. .[.6. The antisense oligonucleotide of claim 1 comprising 25 nucleotides in length..]. .[.7. The antisense oligonucleotide of claim 1 comprising 17-30 nucleotides in length..]. .[.8. The antisense oligonucleotide of claim 1 comprising 20-31 nucleotides in length..]. .[.9. The antisense oligonucleotide of claim 1, wherein the oligonucleotide does not activate RNase H..]. .[.10. The antisense oligonucleotide of claim 1, comprising a non-natural backbone..]. .[.11. The antisense oligonucleotide of claim 10, wherein the sugar moieties of the oligonucleotide backbone are replaced with non-natural moieties..]. .[.12. The antisense oligonucleotide of claim 11, wherein the non-natural moieties are morpholinos..]. .[.13. The antisense oligonucleotide of claim 1, wherein the inter-nucleotide linkages of the oligonucleotide backbone are replaced with non-natural inter-nucleotide linkages..]. .[.14. The antisense oligonucleotide of claim 13, wherein the non-natural inter-nucleotide linkages are modified phosphates..]. .[.15. The antisense oligonucleotide of claim 1, wherein the sugar moieties of the oligonucleotide backbone are replaced with non-natural moieties and the inter-nucleotide linkages of the oligonucleotide backbone are replaced with non-natural inter-nucleotide linkages..]. .[.16. The antisense oligonucleotide of claim 15, wherein the non-natural moieties are morpholinos and the non-natural internucleotide linkages are modified phosphates..]. .[.17. The antisense oligonucleotide of claim 16, wherein the modified phosphates are methyl phosphonates, methyl phosphorothioates, phosphoromorpholidates, phosphoropiperazidates or phosphoroamidates..]. .[.18. The antisense oligonucleotide of claim 1, wherein the oligonucleotide is a 2'-O-methyl-oligoribonucleotide..]. .[.19. The antisense oligonucleotide of claim 1, wherein the oligonucleotide is a peptide nucleic acid..]. 20. The antisense oligonucleotide of claim 1, wherein the oligonucleotide is chemically linked to one or more moieties or conjugates that enhance the activity, cellular distribution, or cellular uptake of the antisense oligonucleotide. .[.21. The antisense oligonucleotide of claim 20, wherein the oligonucleotide is conjugated to a polyamine..]. 22. The antisense oligonucleotide of claim 20, wherein the oligonucleotide is chemically linked to a polyethylene glycol chain. .[.23. An isolated antisense oligonucleotide of 20 to 50 nucleotides in length comprising at least 17 consecutive nucleotides complementary to an exon 45 target region of a human dystrophin gene designated as annealing site H45A(-06+20), wherein the antisense oligonucleotide specifically hybridizes to the acceptor splice site inducing exon 45 skipping, and wherein uracil bases in the antisense oligonucleotide are optionally thymine bases..]. .[.24. The antisense oligonucleotide of claim 23, wherein the uracil bases are thymine bases..]. .[.25. The antisense oligonucleotide of claim 23 comprising 25 nucleotides in length..]. .[.26. The antisense oligonucleotide of claim 23 comprising 17-30 nucleotides in length..]. .[.27. The antisense oligonucleotide of claim 23 comprising 20-31 nucleotides in length..]. .[.28. The antisense oligonucleotide of claim 23 comprising at least 17 consecutive nucleotides 100% complementary to the exon 45 target region..]. .[.29. The antisense oligonucleotide of claim 23 comprising 20-31 nucleotides in length and at least 17 consecutive nucleotides 100% complementary to the exon 45 target region..]. .[.30. The antisense oligonucleotide of claim 23 comprising at least 20 consecutive nucleotides complementary to the exon 45 target region..]. .[.31. The antisense oligonucleotide of claim 23 comprising at least 20 consecutive nucleotides 100% complementary to the exon 45 target region..]. .[.32. The antisense oligonucleotide of claim 23 comprising 20-31 nucleotides in length and at least 20 consecutive nucleotides complementary to the exon 45 target region..]. .[.33. The antisense oligonucleotide of claim 23, wherein the oligonucleotide does not activate RNase H..]. .[.34. The antisense oligonucleotide of claim 23, comprising a non-natural backbone..]. .[.35. The antisense oligonucleotide of claim 34, wherein the sugar moieties of the oligonucleotide backbone are replaced with non-natural moieties..]. .[.36. The antisense oligonucleotide of claim 35, wherein the non-natural moieties are morpholinos..]. .[.37. The antisense oligonucleotide of claim 23, wherein the inter-nucleotide linkages of the oligonucleotide backbone are replaced with non-natural inter-nucleotide linkages..]. .[.38. The antisense oligonucleotide of claim 37, wherein the non-natural inter-nucleotide linkages are modified phosphates..]. .[.39. The antisense oligonucleotide of claim 23, wherein the sugar moieties of the oligonucleotide backbone are replaced with non-natural moieties and the inter-nucleotide linkages of the oligonucleotide backbone are replaced with non-natural inter-nucleotide linkages..]. .[.40. The antisense oligonucleotide of claim 39, wherein the non-natural moieties are morpholinos and the non-natural internucleotide linkages are modified phosphates..]. .[.41. The antisense oligonucleotide of claim 40, wherein the modified phosphates are methyl phosphonates, methyl phosphorothioates, phosphoromorpholidates, phosphoropiperazidates or phosphoroamidates..]. .[.42. The antisense oligonucleotide of claim 23, wherein the oligonucleotide is a 2'-O-methyl-oligoribonucleotide..]. .[.43. The antisense oligonucleotide of claim 23, wherein the oligonucleotide is a peptide nucleic acid..]. .[.44. The antisense oligonucleotide of claim 23, wherein the oligonucleotide is chemically linked to one or more moieties or conjugates that enhance the activity, cellular distribution, or cellular uptake of the antisense oligonucleotide..]. .[.45. The antisense oligonucleotide of claim 44, wherein the oligonucleotide is conjugated to a polyamine..]. .[.46. The antisense oligonucleotide of claim 44, wherein the oligonucleotide is chemically linked to a polyethylene glycol chain..]. .[.47. An isolated antisense oligonucleotide of 20 to 50 nucleotides in length comprising at least 20 consecutive nucleotides of SEQ ID NO: 207, wherein the uracil bases are optionally thymine bases..]. .[.48. The antisense oligonucleotide of claim 47, wherein the uracil bases are thymine bases..]. .[.49. The antisense oligonucleotide of claim 47 comprising 25 nucleotides in length..]. .[.50. The antisense oligonucleotide of claim 47 comprising 20-31 nucleotides in length..]. .[.51. The antisense oligonucleotide of claim 47, wherein the oligonucleotide does not activate RNase H..]. .[.52. The antisense oligonucleotide of claim 47, comprising a non-natural backbone..]. .[.53. The antisense oligonucleotide of claim 52, wherein the sugar moieties of the oligonucleotide backbone are replaced with non-natural moieties..]. .[.54. The antisense oligonucleotide of claim 53, wherein the non-natural moieties are morpholinos..]. .[.55. The antisense oligonucleotide of claim 47, wherein the inter-nucleotide linkages of the oligonucleotide backbone are replaced with non-natural inter-nucleotide linkages..]. .[.56. The antisense oligonucleotide of claim 55, wherein the non-natural inter-nucleotide linkages are modified phosphates..]. .[.57. The antisense oligonucleotide of claim 47, wherein the sugar moieties of the oligonucleotide backbone are replaced with non-natural moieties and the inter-nucleotide linkages of the oligonucleotide backbone are replaced with non-natural inter-nucleotide linkages..]. .[.58. The antisense oligonucleotide of claim 57, wherein the non-natural moieties are morpholinos and the non-natural internucleotide linkages are modified phosphates..]. .[.59. The antisense oligonucleotide of claim 58, wherein the modified phosphates are methyl phosphonates, methyl phosphorothioates, phosphoromorpholidates, phosphoropiperazidates or phosphoroamidates..]. .[.60. The antisense oligonucleotide of claim 47, wherein the oligonucleotide is a 2'-O-methyl-oligoribonucleotide..]. .[.61. The antisense oligonucleotide of claim 47, wherein the oligonucleotide is a peptide nucleic acid..]. .[.62. The antisense oligonucleotide of claim 47, wherein the oligonucleotide is chemically linked to one or more moieties or conjugates that enhance the activity, cellular distribution, or cellular uptake of the antisense oligonucleotide..]. .[.63. The antisense oligonucleotide of claim 62, wherein the oligonucleotide is conjugated to a polyamine..]. .[.64. The antisense oligonucleotide of claim 62, wherein the oligonucleotide is chemically linked to a polyethylene glycol chain..]. 65. A pharmaceutical composition comprising an .Iadd.isolated .Iaddend.antisense oligonucleotide of claim 1, and a pharmaceutically acceptable carrier. .[.66. A pharmaceutical composition comprising an antisense oligonucleotide of claim 23, and a pharmaceutically acceptable carrier..]. .[.67. A pharmaceutical composition comprising an antisense oligonucleotide of claim 47, and a pharmaceutically acceptable carrier..]. 68. A method of inducing exon-skipping of dystrophin exon 45, comprising administering a pharmaceutical composition of claim 65. 69. A method of treating Duchenne muscular dystrophy, comprising administering to a patient in need thereof an effective amount of a pharmaceutical composition of claim 65. .Iadd.70. An injectable solution comprising: an antisense oligonucleotide of 20 to 31 bases comprising a base sequence that is 100% complementary to consecutive bases of exon 45 of the human dystrophin pre-mRNA, wherein the base sequence comprises at least 20 consecutive bases of CCA AUG CCA UCC UGG AGU UCC UGU AA (SEQ ID NO: 207), in which uracil bases are thymine bases, wherein the antisense oligonucleotide is a morpholino antisense oligonucleotide, and wherein the antisense oligonucleotide induces exon 45 skipping; and a pharmaceutically acceptable carrier or diluent; wherein the injectable solution is formulated for intravenous administration..Iaddend. .Iadd.71. The injectable solution of claim 70, wherein the pharmaceutically acceptable carrier or diluent comprises a saline solution..Iaddend. .Iadd.72. The injectable solution of claim 70, wherein the antisense oligonucleotide is chemically linked to a polyethylene glycol chain..Iaddend. .Iadd.73. A method of inducing exon-skipping of dystrophin exon 45, comprising administering the injectable solution of claim 70..Iaddend. .Iadd.74. A method of treating Duchenne muscular dystrophy, comprising administering to a patient in need thereof the injectable solution of claim 70..Iaddend. .Iadd.75. A method of treating Duchenne muscular dystrophy, comprising administering to a patient in need thereof an effective amount of an isolated antisense oligonucleotide of claim 1..Iaddend. .Iadd.76. A method of inducing exon-skipping of dystrophin exon 45, comprising administering an effective amount of an isolated antisense oligonucleotide of claim 1..Iaddend.