Method for efficient exon (44) skipping in duchenne muscular dystrophy and associated means

• US 10,246,707 B2
• Filed: 09/21/2015
• Issued: 04/02/2019
• Est. Priority Date: 05/14/2008
• Status: Active Grant

• Current Assignee: BioMarin Technologies B.V.
• Original Assignee: Academisch Ziekenhuis Leiden, BioMarin Technologies B.V.
• Inventors: Platenburg, Gerard Johannes, de Kimpe, Josephus Johannes, van Deutekom, Judith Christina Theodora, van Ommen, Garrit-Jan Boudewijn, Aartsma-Rus, Annemieke
• Primary Examiner(s): Shin, Dana H
• Application Number:
  US14/859,598
• Publication Number:
  US 20160053262A1
• Time in Patent Office: 1,289 Days
• 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 21/04 for myasthenia gravis
• 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 2310/111 spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/314 Phosphoramidates
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3513 Protein; Peptide
• CPC Class Codes: C12N 2310/3517 Marker; Tag
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2320/33 Alteration of splicing

Abstract

The invention relates to a nucleic acid molecule that binds and/or is complementary to the nucleotide molecule having sequence 5′-GUGGCUAACAGAAGCU (SEQ ID NO 1) and to its use in a method for inducing skipping of exon 44 of the DMD gene in a DMD patient.

Claims

1. A single stranded antisense oligonucleotide 16 to 25 nucleotides in length that comprises a base sequence of the sequence selected from the group consisting of SEQ ID NO:

5 to SEQ ID NO;

16, SEQ ID NO;

20 to SEQ ID NO;

28, SEQ ID NO;

30 to SEQ ID NO;

34, SEQ ID NO;

41, and SEQ ID NO;

46, wherein the antisense oligonucleotide induces skipping of exon 44 of human dystrophin pre-mRNA, and comprises a modification.

2. The antisense oligonucleotide according to claim 1, wherein said antisense oligonucleotide comprises the base sequence of the sequence SEQ ID NO:

5.

3. The antisense oligonucleotide according to claim 1, comprising a 2′

-O-alkyl phosphorothioate antisense oligonucleotide.

4. The antisense oligonucleotide according to claim 3, wherein said antisense oligonucleotide comprises a 2′

-O-methyl phosphorothioate ribose.

5. A viral-based vector, comprising a Pol III-promoter driven expression cassette for expression of an antisense oligonucleotide according to claim 1.

6. The antisense oligonucleotide of claim 1, wherein said modification comprises a modified backbone.

7. The antisense oligonucleotide according to claim 6, wherein the modified backbone is selected from the group consisting of a morpholino backbone, a carbamate backbone, a siloxane backbone, a sulfide backbone, a sulfoxide backbone, a sulfone backbone, a formacetyl backbone, a thioformacetyl backbone, a methyleneformacetyl backbone, a riboacetyl backbone, an alkene containing backbone, a sulfamate backbone, a sulfonate backbone, a sulfonamide backbone, a methyleneimino backbone, a methylenehydrazino backbone and an amide backbone.

8. The antisense oligonucleotide according to claim 1, wherein said modification is selected from the group consisting of:

phosphorodiamidate morpholino oligomer (PMO), peptide nucleic acid, and locked nucleic acid.

9. The antisense oligonucleotide according to claim 2, wherein the antisense oligonucleotide is a PMO.

10. The oligonucleotide of claim 1, wherein said oligonucleotide is conjugated to a ligand.

11. The oligonucleotide of claim 10, wherein said ligand is a peptide.

12. The antisense oligonucleotide according to claim 1, wherein said modification comprises a locked nucleic acid (LNA).