Claims for Patent: RE47691

Summary for Patent: RE47691

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:	15/645,842
Patent Claims:	1. An isolated antisense oligonucleotide of 20 to 50 nucleotides in length comprising at least 20 consecutive nucleotides of SEQ ID NO:193, wherein the oligonucleotide specifically hybridizes to an exon 53 target region of the human dystrophin gene inducing exon 53 skipping, and wherein the uracil bases are optionally thymine bases..]. .[.2. The antisense oligonucleotide of claim 1 comprising SEQ ID NO:193..]. .[.3. The antisense oligonucleotide of claim 1 consisting of SEQ ID NO:193..]. .[.4. The antisense oligonucleotide of claim 1 comprising 20-31 nucleotides in length..]. .[.5. The antisense oligonucleotide of claim 1, wherein the oligonucleotide does not activate RNase H..]. .[.6. The antisense oligonucleotide of claim 1, comprising a non-natural backbone..]. .[.7. The antisense oligonucleotide of claim 1, wherein the sugar moieties of the oligonucleotide backbone are replaced with non-natural moieties..]. .[.8. The antisense oligonucleotide of claim 7, wherein the non-natural moieties are morpholinos..]. .[.9. The antisense oligonucleotide of claim 1, wherein the inter-nucleotide linkages of the oligonucleotide backbone are replaced with non-natural inter-nucleotide linkages..]. .[.10. The antisense oligonucleotide of claim 9, wherein the non-natural inter-nucleotide linkages are modified phosphates..]. .[.11. 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..]. .[.12. The antisense oligonucleotide of claim 11, wherein the non-natural moieties are morpholinos and the non-natural internucleotide linkages are modified phosphates..]. .[.13. The antisense oligonucleotide of claim 12, wherein the modified phosphates are methyl phosphonates, methyl phosphorothioates, phosphoromorpholidates, phosphoropiperazidates or phosphoroamidates..]. .[.14. The antisense oligonucleotide of claim 1, wherein the oligonucleotide is a 2'-O-methyl-oligoribonucleotide..]. .[.15. The antisense oligonucleotide of claim 1, wherein the oligonucleotide is a peptide nucleic acid..]. .[.16. 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..]. .[.17. The antisense oligonucleotide of claim 16, wherein the oligonucleotide is conjugated to a polyamine..]. .[.18. The antisense oligonucleotide of claim 16, wherein the oligonucleotide is chemically linked to a polyethylene glycol chain..]. .[.19. An isolated antisense oligonucleotide of 20 to 50 nucleotides in length comprising at least 20 consecutive nucleotides complementary to an exon 53 target region of the human dystrophin gene designated as annealing site H53A(+39+69), wherein the antisense oligonucleotide specifically hybridizes to the annealing site inducing exon 53 skipping, and wherein uracil bases in the antisense oligonucleotide are optionally thymine bases..]. .[.20. The antisense oligonucleotide of claim 19 comprising 20-31 nucleotides in length..]. .[.21. The antisense oligonucleotide of claim 19, wherein the uracil bases are thymine..]. .[.22. The antisense oligonucleotide of claim 19, wherein the oligonucleotide does not activate RNase H..]. .[.23. The antisense oligonucleotide of claim 19, comprising a non-natural backbone..]. .[.24. The antisense oligonucleotide of claim 19, wherein the sugar moieties of the oligonucleotide backbone are replaced with non-natural moieties..]. .[.25. The antisense oligonucleotide of claim 24, wherein the non-natural moieties are morpholinos..]. .[.26. The antisense oligonucleotide of claim 19, wherein the inter-nucleotide linkages of the oligonucleotide backbone are replaced with non-natural inter-nucleotide linkages..]. .[.27. The antisense oligonucleotide of claim 26, wherein the non-natural inter-nucleotide linkages are modified phosphates..]. .[.28. The antisense oligonucleotide of claim 19, 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..]. .[.29. The antisense oligonucleotide of claim 28, wherein the non-natural moieties are morpholinos and the non-natural internucleotide linkages are modified phosphates..]. .[.30. The antisense oligonucleotide of claim 29, wherein the modified phosphates are methyl phosphonates, methyl phosphorothioates, phosphoromorpholidates, phosphoropiperazidates or phosphoroamidates..]. .[.31. The antisense oligonucleotide of claim 19, wherein the oligonucleotide is a 2'-O-methyl-oligoribonucleotide..]. .[.32. The antisense oligonucleotide of claim 19, wherein the oligonucleotide is a peptide nucleic acid..]. .[.33. The antisense oligonucleotide of claim 19, 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..]. .[.34. The antisense oligonucleotide of claim 33, wherein the oligonucleotide is conjugated to a polyamine..]. .[.35. The antisense oligonucleotide of claim 33, wherein the oligonucleotide is chemically linked to a polyethylene glycol chain..]. .[.36. A pharmaceutical composition, comprising an antisense oligonucleotide of claim 1, and a saline solution that includes a phosphate buffer..]. .[.37. The antisense oligonucleotide of claim 1, wherein the uracil bases are thymine bases..]. .[.38. The antisense oligonucleotide of claim 1, comprising SEQ ID NO:193, wherein the uracil bases are thymine bases..]. .[.39. A pharmaceutical composition, comprising an antisense oligonucleotide of claim 2, and a saline solution that includes a phosphate buffer..]. .[.40. A pharmaceutical composition, comprising an antisense oligonucleotide of claim 3, and a saline solution that includes a phosphate buffer..]. .[.41. A pharmaceutical composition, comprising an antisense oligonucleotide of claim 19, and a saline solution that includes a phosphate buffer..]. .[.42. A pharmaceutical composition, comprising an antisense oligonucleotide of claim 38, and a saline solution that includes a phosphate buffer..]. .[.43. A method of treating Duchenne muscular dystrophy, comprising administering to a patient in need thereof an effective amount of a pharmaceutical composition of claim 36..]. .Iadd.44. An injectable solution comprising: a morpholino antisense oligonucleotide 25 bases in length comprising a base sequence that is 100% complementary to 25 consecutive nucleotides of a target region of exon 53 of the human dystrophin pre-mRNA, said morpholino antisense oligonucleotide chemically linked to a polyethylene glycol chain; wherein the antisense oligonucleotide base sequence comprises at least 20 consecutive bases of C AUU CAA CUG UUG CCU CCG GUU CUG AAG GUG (SEQ ID NO: 193), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping; and a pharmaceutically acceptable carrier or diluent; wherein the injectable solution is formulated for intravenous administration. .Iaddend. .Iadd.45. The injectable solution of claim 44, wherein the pharmaceutically acceptable carrier or diluent comprises an isotonic saline solution. .Iaddend. .Iadd.46. An injectable solution comprising: a morpholino antisense oligonucleotide 25 bases in length comprising a base sequence that is 100% complementary to 25 consecutive nucleotides of a target region of exon 53 of the human dystrophin pre-mRNA, said morpholino antisense oligonucleotide chemically linked to a polyethylene glycol chain; wherein the antisense oligonucleotide base sequence comprises at least 20 consecutive bases of C AUU CAA CUG UUG CCU CCG GUU CUG AAG GUG (SEQ ID NO: 193), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping; and phosphate-buffered saline; wherein the injectable solution is formulated for intravenous administration. .Iaddend. .Iadd.47. An injectable solution comprising: a morpholino antisense oligonucleotide 25 bases in length comprising a base sequence that is 100% complementary to 25 consecutive nucleotides of a target region of exon 53 of the human dystrophin pre-mRNA, said morpholino antisense oligonucleotide chemically linked to a polyethylene glycol chain; wherein the antisense oligonucleotide base sequence comprises at least 20 consecutive bases of C AUU CAA CUG UUG CCU CCG GUU CUG AAG GUG (SEQ ID NO: 193), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping; and a pharmaceutically acceptable carrier or diluent; wherein the injectable solution is formulated for parenteral administration. .Iaddend. .Iadd.48. The injectable solution of claim 47, wherein the pharmaceutically acceptable carrier or diluent comprises an isotonic saline solution. .Iaddend. .Iadd.49. An injectable solution comprising: a morpholino antisense oligonucleotide 25 bases in length comprising a base sequence that is 100% complementary to 25 consecutive nucleotides of a target region of exon 53 of the human dystrophin pre-mRNA, said morpholino antisense oligonucleotide chemically linked to a polyethylene glycol chain; wherein the antisense oligonucleotide base sequence comprises at least 20 consecutive bases of C AUU CAA CUG UUG CCU CCG GUU CUG AAG GUG (SEQ ID NO: 193), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping; and phosphate-buffered saline; wherein the injectable solution is formulated for parenteral administration. .Iaddend. .Iadd.50. An injectable solution comprising: a morpholino antisense oligonucleotide 25 bases in length comprising a base sequence that is 100% complementary to 25 consecutive nucleotides of a target region of exon 53 of the human dystrophin pre-mRNA, said morpholino antisense oligonucleotide chemically linked to a polyethylene glycol chain; wherein the antisense oligonucleotide base sequence comprises at least 20 consecutive bases of C AUU CAA CUG UUG CCU CCG GUU CUG AAG GUG (SEQ ID NO: 193), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping: and a pharmaceutically acceptable carrier or diluent: wherein the injectable solution is formulated for intramuscular administration. .Iaddend. .Iadd.51. The injectable solution of claim 50, wherein the pharmaceutically acceptable carrier or diluent comprises an isotonic saline solution. .Iaddend. .Iadd.52. An injectable solution comprising: a morpholino antisense oligonucleotide 25 bases in length comprising a base sequence that is 100% complementary to 25 consecutive nucleotides of a target region of exon 53 of the human dystrophin pre-mRNA, said morpholino antisense oligonucleotide chemically linked to a polyethylene glycol chain; wherein the antisense oligonucleotide base sequence comprises at least 20 consecutive bases of C AUU CAA CUG UUG CCU CCG GUU CUG AAG GUG (SEQ ID NO: 193), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping; and phosphate-buffered saline; wherein the injectable solution is formulated for intramuscular administration. .Iaddend. .Iadd.53. An injectable solution comprising: a morpholino antisense oligonucleotide 25 bases in length comprising a base sequence that is 100% complementary to 25 consecutive nucleotides of a target region of exon 53 of the human dystrophin pre-mRNA, said morpholino antisense oligonucleotide chemically linked to a polyethylene glycol chain; wherein the antisense oligonucleotide base sequence comprises at least 20 consecutive bases of C AUU CAA CUG UUG CCU CCG GUU CUG AAG GUG (SEQ ID NO: 193), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping; and a pharmaceutically acceptable carrier or diluent; wherein the injectable solution is formulated for subcutaneous administration. .Iaddend. .Iadd.54. The injectable solution of claim 53, wherein the pharmaceutically acceptable carrier or diluent comprises an isotonic saline solution. .Iaddend. .Iadd.55. An injectable solution comprising: a morpholino antisense oligonucleotide 25 bases in length comprising a base sequence that is 100% complementary to 25 consecutive nucleotides of a target region of exon 53 of the human dystrophin pre-mRNA, said morpholino antisense oligonucleotide chemically linked to a polyethylene glycol chain; wherein the antisense oligonucleotide base sequence comprises at least 20 consecutive bases of C AUU CAA CUG UUG CCU CCG GUU CUG AAG GUG (SEQ ID NO: 193), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping; and phosphate-buffered saline; wherein the injectable solution is formulated for subcutaneous administration. .Iaddend. .Iadd.56. An injectable solution comprising: a morpholino antisense oligonucleotide 25 bases in length comprising a base sequence that is 100% complementary to 25 consecutive nucleotides of a target region of exon 53 of the human dystrophin pre-mRNA, said morpholino antisense oligonucleotide chemically linked to a polyethylene glycol chain; wherein the antisense oligonucleotide base sequence comprises at least 20 consecutive bases of C AUU CAA CUG UUG CCU CCG GUU CUG AAG GUG (SEQ ID NO: 193), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping; and a pharmaceutically acceptable carrier or diluent; wherein the injectable solution is formulated for intraocular administration. .Iaddend. .Iadd.57. The injectable solution of claim 56, wherein the pharmaceutically acceptable carrier or diluent comprises an isotonic saline solution. .Iaddend. .Iadd.58. An injectable solution comprising: a morpholino antisense oligonucleotide 25 bases in length comprising a base sequence that is 100% complementary to 25 consecutive nucleotides of a target region of exon 53 of the human dystrophin pre-mRNA, said morpholino antisense oligonucleotide chemically linked to a polyethylene glycol chain; wherein the antisense oligonucleotide base sequence comprises at least 20 consecutive bases of C AUU CAA CUG UUG CCU CCG GUU CUG AAG GUG (SEQ ID NO: 193), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping; and phosphate-buffered saline; wherein the injectable solution is formulated for intraocular administration. .Iaddend. .Iadd.59. An injectable solution comprising: a morpholino antisense oligonucleotide 25 bases in length comprising a base sequence that is 100% complementary to 25 consecutive nucleotides of a target region of exon 53 of the human dystrophin pre-mRNA, said morpholino antisense oligonucleotide chemically linked to a polyethylene glycol chain; wherein the antisense oligonucleotide base sequence comprises at least 20 consecutive bases of C AUU CAA CUG UUG CCU CCG GUU CUG AAG GUG (SEQ ID NO: 193), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping; and a pharmaceutically acceptable carrier or diluent; wherein the injectable solution is formulated for oral administration. .Iaddend. .Iadd.60. The injectable solution of claim 59, wherein the pharmaceutically acceptable carrier or diluent comprises an isotonic saline solution. .Iaddend. .Iadd.61. An injectable solution comprising: a morpholino antisense oligonucleotide 25 bases in length comprising a base sequence that is 100% complementary to 25 consecutive nucleotides of a target region of exon 53 of the human dystrophin pre-mRNA, said morpholino antisense oligonucleotide chemically linked to a polyethylene glycol chain; wherein the antisense oligonucleotide base sequence comprises at least 20 consecutive bases of C AUU CAA CUG UUG CCU CCG GUU CUG AAG GUG (SEQ ID NO: 193), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping; and phosphate-buffered saline; wherein the injectable solution is formulated for oral administration. .Iaddend. .Iadd.62. An injectable solution comprising: a morpholino antisense oligonucleotide 25 bases in length comprising a base sequence that is 100% complementary to 25 consecutive nucleotides of a target region of exon 53 of the human dystrophin pre-mRNA, said morpholino antisense oligonucleotide chemically linked to a polyethylene glycol chain; wherein the antisense oligonucleotide base sequence comprises at least 20 consecutive bases of C AUU CAA CUG UUG CCU CCG GUU CUG AAG GUG (SEQ ID NO: 193), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping; and a pharmaceutically acceptable carrier or diluent; wherein the injectable solution is formulated for transdermal administration. .Iaddend. .Iadd.63. The injectable solution of claim 62, wherein the pharmaceutically acceptable carrier or diluent comprises an isotonic saline solution. .Iaddend. .Iadd.64. An injectable solution comprising: a morpholino antisense oligonucleotide 25 bases in length comprising a base sequence that is 100% complementary to 25 consecutive nucleotides of a target region of exon 53 of the human dystrophin pre-mRNA, said morpholino antisense oligonucleotide chemically linked to a polyethylene glycol chain; wherein the antisense oligonucleotide base sequence comprises at least 20 consecutive bases of C AUU CAA CUG UUG CCU CCG GUU CUG AAG GUG (SEQ ID NO: 193), in which the uracil bases are thymine bases, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping; and phosphate-buffered saline; wherein the injectable solution is formulated for transdermal administration. .Iaddend.