METHODS AND MEANS FOR EFFICIENT SKIPPING OF EXON 45 IN DUCHENNE MUSCULAR DYSTROPHY PRE-mRNA

• US 20120022134A1
• Filed: 04/26/2011
• Published: 01/26/2012
• Est. Priority Date: 10/26/2007
• Status: Active Grant

• Current Assignee: Academisch Ziekenhuis Leiden, BioMarin Technologies B.V.
• Original Assignee: Academisch Ziekenhuis Leiden, Prosensa B.V., Prosensa Holding B.V., Prosensa Technologies B.V.
• Inventors: VAN DEUTEKOM, JUDITH CHRISTINA THEODORA, DE KIMPE, JOSEPHUS JOHANNES, PLATENBURG, GERARDUS JOHANNES, AARTSMA-RUS, ANNEMIEKE, VAN OMMEN, GARRIT-JAN BOUDEWIJN
• Granted Patent: US 9,926,557 B2
• Time in Patent Office: Days
• US Class Current: 514/44.A
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/56 Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
• CPC Class Codes: A61K 31/57 substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
• CPC Class Codes: A61K 31/573 substituted in position 21, e.g. cortisone, dexamethasone, prednisone or aldosterone
• CPC Class Codes: A61K 31/58 containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin digitoxin A61K31/7048
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 38/1719 Muscle proteins, e.g. myosin or actin
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• 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 21/02 Muscle relaxants, e.g. for tetanus or cramps
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• 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: A61P 3/14 for calcium homeostasis vitamin D A61P3/02; parathyroid hormones A61P5/18; calcitonin A61P5/22; osteoporosis A61P19/10; bone metastasis A61P35/04
• CPC Class Codes: A61P 39/06 Free radical scavengers or antioxidants
• 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 2310/31 : of the backbone
• CPC Class Codes: C12N 2310/313 : Phosphorodithioates
• CPC Class Codes: C12N 2310/314 : Phosphoramidates
• 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 2310/3233 : Morpholino-type ring
• CPC Class Codes: C12N 2310/346 : having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3521 : Methyl
• CPC Class Codes: C12N 2320/31 : Combination therapy
• CPC Class Codes: C12N 2320/33 : Alteration of splicing

Abstract

The invention relates to a method for inducing or promoting skipping of exon 45 of DMD pre-mRNA in a Duchenne Muscular Dystrophy patient, preferably in an isolated (muscle) cell, the method comprising providing said cell with a molecule that binds to a continuous stretch of at least 21 nucleotides within said exon. The invention further relates to such molecule used in said method.

Claims

1. A molecule that binds to a continuous stretch of at least 21 nucleotides within exon 45 of DMD pre-mRNA.

2. A molecule according to claim 1, whereby said molecule binds to a continuous stretch of at least 25 nucleotides within said exon.

3. A molecule according to claim 1, whereby said molecule comprises an antisense oligonucleotide of between 21 and 30 bases.

4. A molecule according to claim 1, whereby said molecule comprises an antisense oligonucleotide of 25 bases.

5. A molecule according to claim 1, whereby said molecule binds to a continuous stretch of at least 21 nucleotides within the following nucleotide sequence:

6. A molecule according to claim 1 comprising or consisting of an antisense nucleotide sequence selected from the sequences presented in Table 1, wherein the sequence is not SEQ ID NO:

68.

7. A molecule according to claim 6, comprising or consisting an antisense nucleotide sequence selected from SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 5, SEQ ID NO 6, SEQ ID NO 7 and/or SEQ ID NO 8.

8. A molecule according to claim 7, comprising or consisting of the antisense nucleotide sequence presented in SEQ ID NO 3.

9. A molecule according to claim 8, comprising a 2′

-O-alkyl phosphorothioate antisense oligonucleotide.

10. A molecule according to claim 9, comprising a 2′

-O methyl phosphorothioate ribose.

11. A viral-based vector, comprising an expression cassette that drives expression of the molecule as defined in claim 1.

12. A pharmaceutical composition comprising a molecule as defined in claim 1, a pharmaceutical acceptable carrier, and optionally combined with a molecule which is able to induce or promote skipping of exon 7, 44, 46, 51, 53, 59, 67 (as depicted in table 2) of the DMD pre-mRNA of a patient.

13. A method of treating a patient with DMD comprising administering the molecule of claim 1, and inducing or promoting skipping of exon 45 in the DMD pre-mRNA, and optionally in combination with skipping of one of the exons selected from the group consisting of exon 7, 44, 46, 51, 53, 59 and 67, thereby treating said patient.

14. A method for inducing or promoting skipping of exon 45 of DMD pre-mRNA in a patient, preferably in an isolated cell of said patient, the method comprising providing said cell with a molecule that binds to a continuous stretch of at least 21 nucleotides within said exon.

15. A method according to claim 14, wherein the patient is provided with a functional dystrophin protein and/or wherein the production of an aberrant dystrophin protein in said patient is decreased, wherein the level of said functional dystrophin is assessed by comparison to the level of said dystrophin in said patient at the onset of the method.