Oligonucleotide comprising an inosine for treating DMD

• US 10,533,171 B2
• Filed: 03/24/2017
• Issued: 01/14/2020
• Est. Priority Date: 04/24/2009
• Status: Active Grant

• Current Assignee: BioMarin Technologies B.V.
• Original Assignee: BioMarin Technologies B.V.
• Inventors: Van Deutekom, Judith Christina Theodora, De Kimpe, Josephus Johannes, Platenburg, Gerard Johannes
• Primary Examiner(s): Chong, Kimberly
• Application Number:
  US15/468,239
• Publication Number:
  US 20170204414A1
• Time in Patent Office: 1,026 Days
• Field of Search: None
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• 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 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/3181 Peptide nucleic acid, PNA
• 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 2310/331 Universal or degenerate base
• CPC Class Codes: C12N 2310/333 Modified A
• CPC Class Codes: C12N 2310/336 Modified G
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2320/33 Alteration of splicing

Abstract

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.

Claims

1. An isolated antisense oligonucleotide consisting of the base sequence of SEQ ID NO:

557, provided that said isolated antisense oligonucleotide comprises at least one position wherein a guanosine base is substituted with an inosine base, wherein said isolated antisense oligonucleotide comprises a modification and induces skipping of said exon 45 of human dystrophin pre-mRNA.

2. The isolated antisense oligonucleotide of claim 1, said oligonucleotide comprising from one to four inosine bases.

3. The isolated antisense oligonucleotide of claim 1, wherein the modification is a base and/or sugar modification.

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

-O-methyl phosphorothioate oligonucleotide.

5. The isolated antisense oligonucleotide of claim 1, which is a locked nucleic acid (LNA) oligonucleotide.

6. The isolated antisense oligonucleotide of claim 1, which is a peptide nucleic acid (PNA) oligonucleotide.

7. The isolated antisense oligonucleotide of claim 1, which is a phosphorodiamidate morpholino oligomer (PMO) oligonucleotide.

8. The isolated antisense oligonucleotide of claim 1, wherein the modification is a modified internucleoside linkage.

9. A pharmaceutical composition comprising the isolated antisense oligonucleotide of claim 1 and a pharmaceutically acceptable carrier.

10. A method for inducing skipping of exon 45 of human dystrophin pre-mRNA in a muscle cell, the method comprising contacting said cell with an isolated antisense oligonucleotide of claim 1 for a time and under conditions which permit exon skipping.

11. A method for inducing skipping of exon 45 of human dystrophin pre-mRNA in a human subject, the method comprising administering an isolated antisense oligonucleotide of claim 1 to said subject in an amount and for a time which is effective to induce exon skipping.

12. 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 isolated antisense oligonucleotide of claim 1.