RNA modulating oligonucleotides with improved characteristics for the treatment of duchenne and becker muscular dystrophy

• US 10,179,912 B2
• Filed: 08/09/2016
• Issued: 01/15/2019
• Est. Priority Date: 01/27/2012
• Status: Active Grant

• Current Assignee: BioMarin Technologies B.V.
• Original Assignee: BioMarin Technologies B.V.
• Inventors: de Visser, Peter Christian, van Deutekom, Judith Christina Theodora
• Primary Examiner(s): Schnizer, Richard A
• Application Number:
  US15/232,493
• Publication Number:
  US 20170029818A1
• Time in Patent Office: 889 Days
• Field of Search: None
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3231 having an additional ring, e.g. LNA, ENA
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/331 Universal or degenerate base
• CPC Class Codes: C12N 2310/3341 5-Methylcytosine
• CPC Class Codes: C12N 2310/335 Modified T or U
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2320/33 Alteration of splicing

Abstract

The current invention provides an improved oligonucleotide and its use for treating, ameliorating, preventing and/or delaying DMD or BMD.

Claims

1. An isolated antisense oligonucleotide 16-22 nucleotides in length, wherein the oligonucleotide comprises a sequence which is complementary without mismatches to at least 15 contiguous nucleotides of an exon-internal sequence of exon 51 of a human dystrophin pre-mRNA, wherein the exon-internal sequence is the complement of SEQ ID NO:

111, and wherein all cytosines in the antisense oligonucleotide are 5-methylcytosines and all uracils in the antisense oligonucleotide are 5-methyluracils, and wherein the antisense oligonucleotide is capable of inducing skipping of exon 51 of a human dystrophin pre-mRNA.

2. The isolated antisense oligonucleotide of claim 1, wherein the isolated antisense oligonucleotide comprises a peptide nucleic acid, a locked nucleic acid, or a morpholino phosphorodiamidate modification, or a combination thereof.

3. The isolated antisense oligonucleotide of claim 1, wherein the isolated antisense oligonucleotide comprises a locked nucleic acid modification.

4. The isolated antisense oligonucleotide of claim 1, said oligonucleotide being RNA.

5. The isolated antisense oligonucleotide of claim 1, wherein said oligonucleotide is at least 16 nucleotides in length and comprises a locked nucleic acid modification.

6. 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.

7. 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.

8. A method for treating 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.

9. A method for treating one or more symptom(s) of Duchenne Muscular Dystrophy or Becker Muscular Dystrophy in a cell, the method comprising administering to said cell one or more isolated oligonucleotides of claim 1.