DYSTROPHIN EXON SKIPPING OLIGONUCLEOTIDES
Provided herein are antisense oligonucleotides (ASOs) that induce skipping of exon 53 of human dystrophin pre-mRNA, and pharmaceutically acceptable derivatives thereof. Also provided are pharmaceutical compositions containing the ASOs and methods of using the ASOs and compositions for treating a subject with Duchenne muscular dystrophy.
1 . An ASO comprising or consisting of the sequence of any one of SEQ ID NO:1-1436 which contains at least one modification.
2 . The ASO of claim 1 wherein all nucleotides are RNA.
3 . The ASO of claim 1 , wherein the chemical modification of the sugar moiety is 2′-MOE, 2′-OMe, 2′-F and/or LNA.
4 . The ASO of claim 1 , wherein the modification is a chemical modification of the sugar moiety of all nucleotides in the ASO.
5 . The ASO of claim 3 , wherein the chemical modification of the sugar moiety of all nucleotides in the ASO is 2′-MOE.
6 . The ASO of claim 1 , wherein the chemical modification of the sugar moiety is 1, 2, 3 or 4 LNAs, and the remaining nucleotides are 2′-MOE.
7 . The ASO of claim 1 , wherein the chemical modification of the sugar moiety of the two 5′ terminal nucleotide positions of the ASO and the two 3′ terminal nucleotide positions of the ASO are all LNA, and the remaining nucleotides are 2′-MOE.
8 . The ASO of claim 1 , wherein the chemical modification of the sugar moiety of one or more nucleotides in the ASO is a morpholine (PMO, PPMO, PMO-X), a peptide derivative (PNA), a boron-cluster modified PNA, a pyrrolidine-based oxy-peptide nucleic acid (POPNA), a glycol- or glycerol-based nucleic acid (GNA), a threose-based nucleic acid (TNA), an acyclic threoninol-based nucleic acid (aTNA), a cationic morpholino-based oligomer (PMOPlus), an oligonucleotide with integrated bases and backbones (ONIBs), a pyrrolidine-amide oligonucleotides (POMs); or a derivative thereof.
9 . The ASO of claim 1 , wherein the modification is a chemical modification of the base moiety of 1, 2, 3, 4 or all nucleotides in the ASO.
10 . The ASO of claim 1 , wherein all cytosine bases are replaced with 5-methylcytosine.
11 . The ASO of claim 1 , wherein all thymine bases are replaced with uracil.
12 . The ASO of claim 1 , wherein the backbone is a fully phosphorothioate backbone linkage.
13 . The ASO of claim 1 , wherein the ASO comprises a hydroxyalkoxy group at the 5′ terminus of the ASO, the 3′ terminus of the ASO, or at both the 5′ and 3′ ends of the ASO.
14 . The ASO of claim 1 , wherein the ASO comprises a modified hydroxyalkoxy group at the 5′ terminus of the ASO, the 3′ terminus of the ASO, or at both the 5′ and 3′ ends of the ASO.
15 . The ASO of claim 14 , wherein the hydroxyalkoxy group comprises or consists of an ethylene glycol monomer, ethylene glycol oligomer or ethylene glycol polymer (also known as polyethylene glycol, PEG).
16 . The ASO of claim 14 , wherein the hydroxyalkoxy group is a TEG or a HEG.
17 . The ASO of claim 1 , wherein one or two nucleotides are omitted from the 5′ terminus of the ASO, or where one or two nucleotides are omitted from the 3′ terminus of the ASO, or where one nucleotide is omitted from the 5′ terminus of the ASO and one nucleotide is omitted from the 3′ terminus of the ASO.
18 . The ASO of claim 17 that is 16 to 30 nucleotides in length.
19 . The ASO of claim 17 that is 16, 17, 18, 19 or 20 nucleotides in length.
20 . The ASO of claim 17 that is 18 nucleotides in length.
21 . The ASO of claim 1 , that is fully 2′-MOE RNA modified, wherein all cytosines are replaced with 5-methylcytosine, and wherein the backbone is a fully phosphorothioate backbone.
22 . The ASO of claim 1 , wherein all cytosines are replaced with 5-methylcytosine, the two 5′ terminal nucleotides of the ASO are LNA, the two 3′ terminal nucleotides of the ASO are LNA, the remaining nucleotides are 2′-MOE RNA modified, and wherein the backbone is a fully phosphorothioate backbone.
23 . A pharmaceutical composition, comprising the ASO of claim 1 and a pharmaceutically acceptable carrier.
24 . A method of treating a subject having DMD, comprising administering to the subject the ASO of claim 1 .
25 . A method of delaying the onset of DMD in a subject, comprising administering to the subject the ASO of claim 1 .
26 . A method of inducing skipping of exon 53 of human dystrophin pre-mRNA, comprising contacting human dystrophin pre-mRNA with the ASO of claim 1 .