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 oligonucleotide comprising a sequence which is complementary to at least part of a dystrophin pre-mRNA exon or at least part of a non-exon region of a dystrophin pre-mRNA said part being a contiguous stretch comprising at least 8 nucleotides, wherein said oligonucleotide comprises one or both of an inosine nucleotide and a nucleotide containing a base able to form a wobble base pair with a complementary base to which it is paired.
2 . An isolated oligonucleotide according to claim 1 , wherein the contiguous stretch comprises between 13 and 50 nucleotides, of RNA of an exon of a dystrophin pre-mRNA.
3 . An isolated oligonucleotide according to claim 2 , wherein said exon comprises exon 51, 45, 53, 44, 46, 52, 50, 43, 6, 7, 8, 55, 2, 11, 17, 19, 21, 57, 59, 62, 63, 65, 66, 69, and/or 75.
4 . (canceled)
5 . An isolated oligonucleotide according to claim 1 , wherein the oligonucleotide comprises a first part and a second part, wherein said first part comprises least 8, consecutive nucleotides that are complementary to a first exon and wherein said second part comprises at least 8 consecutive nucleotides that are complementary to a second exon in said dystrophin pre-mRNA.
6 . An isolated oligonucleotide according to claim 5 , wherein said first and said second exon are separated in said dystrophin pre-mRNA by at least one exon to which said oligonucleotide is not complementary.
7 . An oligonucleotide according to claim 5 , wherein said first and said second exon are contiguous in said dystrophin pre-mRNA.
8 . (canceled)
9 . (canceled)
10 . (canceled)
11 . A composition comprising at least two distinct oligonucleotides as defined in claim 1 .
12 . A composition according to claim 11 , wherein each said distinct oligonucleotide is dosed, independently, in an amount between 0.5 mg/kg and 10 mg/kg, inclusive.
13 . A composition according to claim 11 in combination with one or more of:
(a) an adjunct compound for reducing inflammation, preferably for reducing muscle tissue inflammation,
(b) an adjunct compound for improving muscle fiber function, integrity and/or survival, and
(c) a compound exhibiting readthrough activity.
14 . (canceled)
15 . 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 a composition as defined in claim 11 .
16 . An isolated oligonucleotide according to claim 2 wherein said contiguous stretch comprises between 14 and 25 nucleotides of RNA of an exon of a dystrophin pre-mRNA.
17 . An isolated oligonucleotide according to claim 1 , wherein the oligonucleotide comprises RNA.
18 . An isolated oligonucleotide according to claim 17 , wherein said RNA comprises a modified ribonucleotide.
19 . An isolated oligonucleotide according to claim 18 , wherein said modified ribonucleotide is a 2′-O-methyl modified ribose (RNA).
20 . An isolated oligonucleotide according to claim 1 , comprising a modified deoxyribose (DNA) base.
21 . An isolated oligonucleotide according to claim 1 , wherein said oligonucleotide comprises a peptide nucleic acid, a locked nucleic acid, a morpholino phosphorodiamidate, or a combination thereof.
22 . An isolated oligonucleotide according to claim 21 , comprising a morpholino phosphorodiamidate.
23 . An isolated oligonucleotide according to claim 5 , wherein said first part and said second part, independently, comprise between 16 and 80 consecutive nucleotides, inclusive.
24 . The composition of claim 11 , admixed with a pharmaceutically acceptable carrier, adjuvant, diluent and/or excipient.
25 . The composition of claim 13 , wherein said adjunct composition for reducing inflammation reduces tissue inflammation.
26 . The method of claim 15 , wherein the composition administered provides the individual with a functional dystrophin protein.
27 . The method of claim 15 , wherein the composition administered decreases the production of an aberrant dystrophin protein.
28 . The method of claim 15 , wherein the composition administered increases the production of a functional or a more functional dystrophin protein.
29 . The method of claim 15 , wherein the composition administered alleviates one or more symptom(s).