RNA Modulating Oligonucleotides with Improved Characteristics for the Treatment of Duchenne and Becker Muscular Dystrophy

• US 20170029818A1
• Filed: 08/09/2016
• Published: 02/02/2017
• Est. Priority Date: 01/27/2012
• Status: Active Grant

• Current Assignee: BioMarin Technologies B.V.
• Original Assignee: BioMarin Technologies B.V.
• Inventors: van DEUTEKOM, Judith Christina Theodora, de Visser, Peter Christian
• Granted Patent: US 10,179,912 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• 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 of length 13-33 nucleotides, wherein the oligonucleotide comprises a sequence which is complementary to a dystrophin pre-mRNA exon, said oligonucleotide being a 2′

-O-methyl ribose and 5′

methylcytosine or 5′

methyluracil modified oligonucleotide, and wherein the antisense oligonucleotide is capable of exon-skipping.

2. The isolated antisense oligonucleotide of claim 1, wherein the oligonucleotide comprises 16-22 nucleotides.

3. The isolated antisense oligonucleotide of claim 1, wherein said oligonucleotide is a 2′

-O-methyl-phosphorothioate oligonucleotide.

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

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

6. The isolated antisense oligonucleotide of claim 1, wherein said exon comprises exon 44, 45, 46, 50, 51, 52, and 55.

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

8. The isolated antisense oligonucleotide of claim 1, wherein the isolated antisense oligonucleotide is a 5-methyl cytosine oligonucleotide.

9. The isolated antisense oligonucleotide of claim 1, wherein the isolated antisense oligonucleotide is a 5-methyl uracil oligonucleotide.

10. The isolated antisense oligonucleotide of claim 1, wherein said oligonucleotide comprises a 2, 6-diaminopurine base.

11. The isolated antisense oligonucleotide of claim 1, wherein said oligonucleotide is at least 16 nucleotides in length and comprises 5-methyl cytosine, a locked nucleic acid, and 2′

-O-methyl-phosphorothioate modification.

12. A pharmaceutical composition comprising at least two distinct isolated antisense oligonucleotides as defined claim 1, wherein said pharmaceutical composition further comprises a pharmaceutically acceptable carrier, adjuvant, diluent and/or excipient.

13. 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 a pharmaceutical composition of claim 12, wherein said composition induces skipping of exons of dystrophin pre-mRNA.

14. 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 a pharmaceutical composition of claim 12, wherein said composition induces skipping of exons of dystrophin pre-mRNA.

15. 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.

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 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.

18. 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.