Antisense oligonucleotides for inducing exon skipping and methods of use thereof
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 214.
1. An antisense oligonucleotide comprising a base sequence 25 bases in length that is 100% complementary to 25 consecutive nucleotide bases of a target region of exon 53 of the human dystrophin pre-mRNA, 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, wherein the antisense oligonucleotide is a morpholino antisense oligonucleotide, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping, or a pharmaceutically acceptable salt thereof.
2. The antisense oligonucleotide of claim 1 , wherein the antisense oligonucleotide is chemically linked to a cholesterol moiety, cholic acid, a thioether, a thiocholesterol, an aliphatic chain, a phospholipid, a polyamine chain, a polyethylene glycol chain, adamantane acetic acid, a palmityl moiety, an octadecylamine moiety, or a hexylamino-carbonyl-oxycholesterol moiety.
3. The antisense oligonucleotide of claim 1 , wherein the antisense oligonucleotide is in a free base form.
4. An injectable solution, comprising: an antisense oligonucleotide comprising a base sequence 25 bases in length that is 100% complementary to 25 consecutive nucleotide bases of a target region of exon 53 of the human dystrophin pre-mRNA, 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, wherein the antisense oligonucleotide is a morpholino antisense oligonucleotide, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent, wherein the injectable solution is formulated for intravenous administration.
5. The injectable solution of claim 4 , wherein the antisense oligonucleotide is chemically linked to a cholesterol moiety, cholic acid, a thioether, a thiocholesterol, an aliphatic chain, a phospholipid, a polyamine chain, a polyethylene glycol chain, adamantane acetic acid, a palmityl moiety, an octadecylamine moiety, or a hexylamino-carbonyl-oxycholesterol moiety.
6. The injectable solution of claim 4 , wherein the pharmaceutically acceptable carrier or diluent comprises an isotonic saline solution.
7. The injectable solution of claim 6 , wherein the isotonic saline solution is phosphate-buffered saline.
8. The injectable solution of claim 4 , wherein the antisense oligonucleotide is in a free base form.
9. An injectable solution, comprising an antisense oligonucleotide comprising a base sequence 25 bases in length that is 100% complementary to 25 consecutive nucleotide bases of a target region of exon 53 of the human dystrophin pre-mRNA, 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, wherein the antisense oligonucleotide is a morpholino antisense oligonucleotide, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent, wherein the injectable solution is formulated for parenteral administration.
10. The injectable solution of claim 9 , wherein the antisense oligonucleotide is chemically linked to a cholesterol moiety, cholic acid, a thioether, a thiocholesterol, an aliphatic chain, a phospholipid, a polyamine chain, a polyethylene glycol chain, adamantane acetic acid, a palmityl moiety, an octadecylamine moiety, or a hexylamino-carbonyl-oxycholesterol moiety.
11. The injectable solution of claim 9 , wherein the pharmaceutically acceptable carrier or diluent comprises an isotonic saline solution.
12. The injectable solution of claim 11 , wherein the isotonic saline solution is phosphate-buffered saline.
13. The injectable solution of claim 9 , wherein the antisense oligonucleotide is in a free base form.
14. The injectable solution of claim 9 , wherein the injectable solution is formulated for intramuscular administration.
15. An injectable solution, comprising an antisense oligonucleotide comprising a base sequence 25 bases in length that is 100% complementary to 25 consecutive nucleotide bases of a target region of exon 53 of the human dystrophin pre-mRNA, 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, wherein the antisense oligonucleotide is a morpholino antisense oligonucleotide, and wherein the antisense oligonucleotide specifically hybridizes to the target region and induces exon 53 skipping, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent, wherein the injectable solution is formulated for subcutaneous administration.
16. The injectable solution of claim 15 , wherein the antisense oligonucleotide is chemically linked to a cholesterol moiety, cholic acid, a thioether, a thiocholesterol, an aliphatic chain, a phospholipid, a polyamine chain, a polyethylene glycol chain, adamantane acetic acid, a palmityl moiety, an octadecylamine moiety, or a hexylamino-carbonyl-oxycholesterol moiety.
17. The injectable solution of claim 15 , wherein the pharmaceutically acceptable carrier or diluent comprises an isotonic saline solution.
18. The injectable solution of claim 17 , wherein the isotonic saline solution is phosphate-buffered saline.
19. The injectable solution of claim 15 , wherein the antisense oligonucleotide is in a free base form.