OLIGONUCLEOTIDE COMPRISING AN INOSINE FOR TREATING DMD
The invention provides an oligonucleotide comprising an inosine, and/or a nucleotide containing a base able to form a wobble base pair or a functional equivalent thereof, wherein the oligonucleotide, or a functional equivalent thereof, comprises a sequence which is complementary to at least part of a dystrophin pre-m RNA exon or at least part of a non-exon region of a dystrophin pre-m RNA said part being a contiguous stretch comprising at least 8 nucleotides. The invention further provides the use of said oligonucleotide for preventing or treating DMD or BMD.
1 . An isolated antisense oligonucleotide 13 to 50 nucleotides in length comprising an inosine base, wherein said oligonucleotide is capable of binding to an exon of the human dystrophin pre-mRNA so as to induce skipping of said exon.
2 . The oligonucleotide of claim 1 , said oligonucleotide being 20-50 nucleotides in length.
3 . The oligonucleotide of claim 1 , said oligonucleotide being 14-25 nucleotides in length.
4 . The oligonucleotide of claim 1 , said oligonucleotide being 20-25 nucleotides in length.
5 . The oligonucleotide of claim 1 , said oligonucleotide comprising from one to four inosine bases.
6 . The oligonucleotide of claim 1 , further comprising a base and/or sugar modification.
7 . The oligonucleotide of claim 1 , wherein said oligonucleotide is a 2′-O-methyl phosphorothioate oligonucleotide.
8 . The oligonucleotide of claim 1 , wherein said exon is selected from the group consisting of 51, 45, 53, 44, 46, 52, 50, 43 and 55.
9 . The oligonucleotide of claim 1 , said oligonucleotide being RNA.
10 . An isolated antisense oligonucleotide consisting of a base or a nucleotide sequence selected from the group consisting of: SEQ ID NO: 2-473, 539-576, wherein said oligonucleotide comprises an inosine base.
11 . The oligonucleotide of claim 8 , further comprising a base and/or sugar modification.
12 . The oligonucleotide of claim 8 , wherein said oligonucleotide is a 2′-O-methyl phosphorothioate oligonucleotide.
13 . The oligonucleotide of claim 8 , said oligonucleotide being RNA.
14 . A method for inducing skipping of an exon of human dystrophin pre-mRNA in a muscle cell, the method comprising contacting said cell with an oligonucleotide of claim 1 for a time and under conditions which permit exon skipping.
15 . The oligonucleotide of claim 1 , which is a locked nucleic acid oligonucleotide (LNA).
16 . A method for inducing skipping of an exon of human dystrophin pre-mRNA in a human subject, the method comprising administering an oligonucleotide of claim 1 to said subject in an amount and for a time which is effective to induce exon skipping.
17 . A method for alleviating one or more symptom(s) of Duchenne Muscular Dystrophy or Becker Muscular Dystrophy in an individual, the method comprising administering to said individual an oligonucleotide of claim 1 , wherein said oligonucleotide induces skipping of an exon of a dystrophin pre-mRNA.
18 . A method for inducing and/or promoting skipping of exon 43, 44, 45, 46, 50, 51, 52 or 53 of the dystrophin pre-mRNA in a patient, the method comprising administering an oligonucleotide of claim 4 to said patient.