Means and method for inducing exon-skipping

• US 8,361,979 B2
• Filed: 03/18/2007
• Issued: 01/29/2013
• Est. Priority Date: 05/19/2006
• Status: Active Grant

• Current Assignee: Academisch Ziekenhuis Leiden
• Original Assignee: Academisch Ziekenhuis Leiden
• Inventors: Aartsma-Rus, Annemieke, Van Ommen, Garrit-Jan Boudewijn, Van Deutekom, Judith Christina T.
• Primary Examiner(s): SCHNIZER, RICHARD A
• Application Number:
  US12/300,629
• Publication Number:
  US 20090269755A1
• Time in Patent Office: 2,144 Days
• Field of Search: None
• US Class Current: 514/44.A
• CPC Class Codes: A61K 48/0016 wherein the nucleic acid is delivered as a 'naked' nucleic acid, i.e. not combined with an entity such as a cationic lipid
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 5/14 of the thyroid hormones, e.g. T3, T4
• CPC Class Codes: A61P 7/04 Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 15/907 in mammalian cells
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• 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

In the present invention means and method are provided for optimising exon-skipping using exon-internal AON. We show that skipping efficiencies are improved by targeting putative splicing regulatory sequences (ESEs) within an exon. Such double targeting may be particularly useful for exons with which efficient skipping was difficult to obtain prior to the invention.

Claims

1. An AON comprising a sequence selected from the group consisting of:

SEQ ID NO;

115 to SEQ ID NO;

118.

2. The AON of claim 1, wherein the AON comprises 14-40 nucleotides.

3. The AON of claim 1, wherein the AON comprises 14-25 nucleotides.

4. The AON of claim 1, wherein the AON comprises one or more modifications selected from the group consisting of:

2′

-O-methyl oligoribonucleotides, phosphorothioate backbone, morpholino phosphorodiamidate DNA (morpholinos), locked nucleic acids (LNA); and

ethylene bridged nucleic acids (ENA).

5. A method of treating a subject in need thereof, wherein said subject comprises Duchenne Muscular Dystrophy, comprising administering the AON of claim 1 to the subject and wherein said AON induces the exclusion of exon 33 or 44 from a DMD pre-mRNA in said subject.

6. A pharmaceutical composition comprising the AON of claim 1 in combination with a suitable carrier.

7. A method of inducing the exclusion of an exon from a pre-mRNA associated with Duchenne Muscular Dystrophy in a subjecting having Duchenne Muscular Dystrophy, said method comprising administering the AON of claim 1 to the subject.