Methods and means for efficient skipping of exon 45 in duchenne muscular dystrophy pre-mRNA

Patent metadata

Field	Value
Publication number	US-9528109-B2
Application number	US-201414542183-A
Country	US
Kind code	B2
Filing date	Nov 14, 2014
Priority date	Oct 26, 2007
Publication date	Dec 27, 2016
Grant date	Dec 27, 2016

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 an isolated muscle cell with a molecule that binds to a continuous stretch of at least 21 nucleotides within the exon. The invention further relates to such molecule used in the method.

First claim

We claim:

1. An antisense oligonucleotide whose base sequence consists of the base sequence of 5 ′UUUGCCGCUGCCCAAUGCCAUCCUG-3′ (SEQ ID: NO: 3), said oligonucleotide comprising a modification.

2. The oligonucleotide of claim 1 , comprising a phosphorodiamidate morpholino oligomer (PMO).

3. The oligonucleotide of claim 1 , wherein said oligonucleotide comprises a locked nucleic acid (LNA).

4. The antisense oligonucleotide of claim 1 wherein the oligonucleotide comprises a modified base.

5. The antisense oligonucleotide of claim 1 wherein the oligonucleotide comprises a modified sugar moiety.

6. The antisense oligonucleotide of claim 1 wherein the oligonucleotide comprises a modified internucleoside linkage.

7. The oligonucleotide of claim 1 wherein said oligonucleotide comprises a phosphorothioate internucleoside linkage and a 2′-O-alkyl substituted ribose moiety.

8. The oligonucleotide of claim 5 , wherein the modified sugar moiety is selected from the group consisting of: a ribose that is mono- or di-substituted at the 2′, 3′, and/or 5′ position.

9. The oligonucleotide of claim 8 , wherein the ribose is a 2′-O-substituted ribose.

10. The oligonucleotide of claim 9 , wherein the ribose is a 2′-O-methyl ribose.

11. The oligonucleotide of claim 6 , said oligonucleotide comprising a modified backbone such that all of internucleoside linkages of said oligonucleotide are modified.

12. The oligonucleotide of claim 11 , said internucleoside linkages comprising phosphorothioate.

13. The oligonucleotide of claim 11 , said internucleoside linkages comprising a phosphorodiamidate morpholino oligomer (PMO).

14. The oligonucleotide of claim 1 said modification comprising a peptide nucleic acid, and/or locked nucleic acid.

15. The oligonucleotide of claim 11 , wherein said 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.

16. The antisense oligonucleotide of claim 1 wherein said oligonucleotide is capable of inducing skipping of exon 45 by at least 50%.

17. The antisense oligonucleotide of claim 16 , wherein said oligonucleotide is capable of inducing skipping of exon 45 by at least 60%.

18. The antisense oligonucleotide of claim 17 , wherein said oligonucleotide is capable of inducing skipping of exon 45 by at least 70%.

19. The antisense oligonucleotide of claim 18 , wherein said oligonucleotide is capable of inducing skipping of exon 45 by at least 80%.

20. The antisense oligonucleotide of claim 19 , wherein said oligonucleotide is capable of inducing skipping of exon 45 by at least 90%.

21. A viral-based vector comprising an expression cassette comprising a nucleotide sequence encoding the oligonucleotide of claim 1 .

22. A pharmaceutical composition comprising the oligonucleotide of claim 1 , and a pharmaceutically acceptable carrier.

23. The pharmaceutical composition of claim 1 , further comprising an antisense oligonucleotide which induces or promotes skipping of exon 7, 44, 46, 51, 53, 59, or 67 of dystrophin pre-mRNA of a patient.

Assignees

• Prosensa Tech B V
• Academisch Ziekenhuis Leiden
• Biomarin Tech Bv

Inventors

• De Kimpe Josephus Johannes
• Rus Adriana Marie
• Platenburg Gerard Johannes
• Van Deutekom Judith Christina Theodora
• Van Ommen Garrit-Jan Boudewijn

Classifications

• A61P39/06

  Free radical scavengers or antioxidants · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P29/00

  Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID] · CPC title

• A61P3/14

  for calcium homeostasis (vitamin D A61P3/02; parathyroid hormones A61P5/18; calcitonin A61P5/22; osteoporosis A61P19/10; bone metastasis A61P35/04) · CPC title

• A61P21/02

  Muscle relaxants, e.g. for tetanus or cramps · CPC title