Means and methods for counteracting muscle disorders

• US 20110294753A1
• Filed: 04/26/2010
• Published: 12/01/2011
• Est. Priority Date: 10/26/2007
• Status: Active Grant

• Current Assignee: Academisch Ziekenhuis Leiden, BioMarin Technologies B.V.
• Original Assignee: Academisch Ziekenhuis Leiden, Prosensa Technologies B.V.
• Inventors: Aartsma-Rus, Annemieke, Van Deutekom, Judith Christina Theodora, Van Ommen, Garrit-Jan Boudewijn, Platenburg, Gerard Johannes, De Kimpe, Josephus Johannes
• Granted Patent: US 9,243,245 B2
• Time in Patent Office: Days
• US Class Current: 514/40
• 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 provides means and methods for alleviating one or more symptom(s) of Duchenne Muscular Dystrophy and/or Becker Muscular Dystrophy. Therapies using compounds for providing patients with functional muscle proteins are combined with at least one adjunct compound for reducing inflammation, preferably for reducing muscle tissue inflammation, and/or at least one adjunct compound for improving muscle fiber function, integrity and/or survival.

Claims

1. A composition comprising:

a first compound for providing an individual with a functional

dystrophin protein; and

a second compound;

wherein said second compound comprises at least one of;

a compound for reducing inflammation, and

a compound for improving muscle fiber function, integrity and/or survival.

2. The composition of claim 1, wherein said inflammation is muscle tissue inflammation.

3. The composition of claim 1, wherein said second compound comprises a steroid selected from the group consisting of a (gluco)corticosteroid, an ACE inhibitor (preferably perindopril), angiotensin II type 1 receptor blocker Losartan, a tumour necrosis factor-alpha (TNFα

) inhibitor, a source of mIGF-1, an antioxidant, an ion channel inhibitor, a protease inhibitor and L-arginine.

4. The composition of claim 1, wherein said second compound enhances exon skipping and/or inhibits spliceosome assembly and/or splicing

5. The composition of claim 1, wherein said second compound comprises the combination of a steroid and an agent that enhances exon skipping and/or inhibits spliceosome assembly and/or splicing.

6. The composition of claim 1, wherein said first compound comprises an oligonucleotide, or a functional equivalent thereof, that decreases the production of an aberrant dystrophin protein in said individual.

7. The composition of claim 1, wherein said first compound comprises an oligonucleotide, or a functional equivalent thereof, that inhibits inclusion of an exon of a dystrophin pre-mRNA into mRNA produced from splicing of said pre-mRNA, wherein the absence of said exon from said mRNA produced from said dystrophin pre-mRNA generates a coding region for a functional dystrophin protein.

8. The composition of claim 1, wherein said first compound comprises an oligonucleotide, or a functional equivalent thereof, comprising a sequence which is complementary to at least part of a dystrophin pre-mRNA exon or at least part of a non-exon region of a dystrophin pre-mRNA said part having at least 13 nucleotides.

9. The composition of claim 1, wherein said first compound suppresses stop codons and is selected from the group consisting of gentamicin, PTC124 or a functional equivalent thereof.

10. The composition of claim 1, wherein said first compound comprises an oligonucleotide, or a functional equivalent thereof, comprising a sequence which is complementary to between 13 and 50 consecutive nucleotides of RNA of an exon of a dystrophin pre-mRNA.

11. The composition of claim 1, wherein said first compound comprises an oligonucleotide, or a functional equivalent thereof, comprising a sequence which is complementary to between 14 and 25 consecutive nucleotides of RNA of an exon of a dystrophin pre-mRNA.

12. The composition of claim 1, wherein said first compound comprises an oligonucleotide, or a functional equivalent thereof, that binds specifically to a regulatory RNA sequence required for the correct splicing of dystrophin exons in a transcript and, wherein said oligonucleotide or functional equivalent thereof is complementary to an intronic splicing enhancer (ISE), an exonic splicing enhancer (ESE), an intronic splicing silencer (ISS) and/or an exonic splicing silencer (ESS).

13. The composition of claim 6, wherein said oligonucleotide comprises RNA and wherein said RNA contains a modification.

14. The composition of claim 13, wherein said modification is selected from the group consisting of a 2′

-O-methyl modified ribose (RNA) or deoxyribose (DNA) modification.

15. The composition of claim 6, wherein said functional equivalent of said oligonucleotide is selected from the group consisting of at least one of a peptide nucleic acid, a locked nucleic acid, morpholino phosphorodiamidate, or any combination thereof.

16. The composition of claim 7, wherein said exon comprises at least one of exon 51, 44, 45, 53, 46, 43, 2, 8, 50 and 52.

17. The composition of claim 6, wherein said oligonucleotide is complementary to at least two exons in a dystrophin pre-mRNA, and comprises at least two parts wherein a first part comprises at least 8 consecutive nucleotides that are complementary to a first of said at least two exons and wherein a second part comprises an oligonucleotide having at least 8 consecutive nucleotides that are complementary to a second exon in said dystrophin pre-mRNA.

18. The composition of claim 17, wherein said first and said second exons are separated in said dystrophin pre-mRNA by at least one exon to which said oligonucleotide is not complementary.

19. The composition of claim 17, wherein said first part comprises between 16-80 nucleotides that are complementary to a first of said at least two exons.

20. The composition of claim 17, wherein said second part comprises between 16-80 nucleotides that are complementary to a second exon in said dystrophin pre-mRNA.

21. 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;

a first compound that provides a functional dystrophin protein, and a second compound,

wherein said second compound comprises at least one of;

a compound that reduces inflammation, and/or a compound that improves muscle fiber function, integrity and/or survival.

22. The method of claim 21, wherein said inflammation is muscle tissue inflammation.

23. The method of claim 21, wherein said second compound comprises a steroid selected from the group consisting of a (gluco)corticosteroid, an ACE inhibitor (preferably perindopril), angiotensin II type 1 receptor blocker Losartan, a tumour necrosis factor-alpha (TNFα

) inhibitor, a source of mIGF-1, an antioxidant, an ion channel inhibitor, a protease inhibitor and L-arginine.

24. The method of claim 21 wherein said second compound enhances exon skipping and/or inhibits spliceosome assembly and/or splicing.

25. The method of claim 21, wherein said second compound comprises the combination of a steroid and an agent that enhances exon skipping and/or inhibits spliceosome assembly and/or splicing.

26. The method of claim 21, wherein said first compound comprises an oligonucleotide, or a functional equivalent thereof, that decreases the production of an aberrant dystrophin protein in said individual.

27. The method of claim 21, wherein said first compound comprises an oligonucleotide, or a functional equivalent thereof, that inhibits inclusion of an exon of a dystrophin pre-mRNA into mRNA produced from splicing of said pre-mRNA, wherein the absence of said exon from said mRNA produced from said dystrophin pre-mRNA generates a coding region for a functional dystrophin protein.

28. The method of claim 21, wherein said first compound comprises an oligonucleotide, or a functional equivalent thereof, comprising a sequence which is complementary to at least part of a dystrophin pre-mRNA exon or at least part of a non-exon region of a dystrophin pre-mRNA said part having at least 13 nucleotides.

29. The method of claim 21, wherein said first compound comprises an oligonucleotide, or a functional equivalent thereof, that binds specifically to a regulatory RNA sequence required for the correct splicing of dystrophin exons in a transcript and, wherein said oligonucleotide or functional equivalent thereof is complementary to an intronic splicing enhancer (ISE), an exonic splicing enhancer (ESE), an intronic splicing silencer (ISS) and/or an exonic splicing silencer (ESS).

30. The method of claim 26, wherein said oligonucleotide comprises RNA and wherein said RNA contains a modification.

31. The method of claim 30, wherein said modification is selected from the group consisting of a 2′

-O-methyl modified ribose (RNA) or deoxyribose (DNA) modification.

32. The method of claim 26 wherein said functional equivalent of said oligonucleotide is selected from the group consisting of at least one of a peptide nucleic acid, a locked nucleic acid, morpholino phosphorodiamidate, or any combination thereof.

33. The method of claim 27, wherein said exon comprises at least one of exon 51, 44, 45, 53, 46, 43, 2, 8, 50 and 52.

34. The method of claim 26, wherein said oligonucleotide is complementary to at least two exons in a dystrophin pre-mRNA, and comprises at least two parts wherein a first part comprises at least 8 consecutive nucleotides that are complementary to a first of said at least two exons and wherein a second part comprises an oligonucleotide having at least 8 consecutive nucleotides that are complementary to a second exon in said dystrophin pre-mRNA.

35. The method of claim 34, wherein said first and said second exons are separated in said dystrophin pre-mRNA by at least one exon to which said oligonucleotide is not complementary.

36. A method for increasing the production of a functional dystrophin protein in a cell, said cell comprising pre-mRNA of a dystrophin gene encoding an aberrant dystrophin protein comprising:

providing said cell with a first compound for inhibiting inclusion of an exon into mRNA produced from splicing of said dystrophin pre-mRNA, and providing said cell with a second compound;

wherein said second compound comprises at least one of;

a compound that reduces inflammation, and/or a compound for improving muscle fiber function, integrity and/or survival,

said method further comprising allowing translation of mRNA produced from splicing of said pre-mRNA.

37. The method of claim 36, wherein said inflammation is muscle tissue inflammation,

38. A pharmaceutical preparation comprising:

a first compound for providing an individual with a functional dystrophin protein, and a second compound, wherein said second compound comprises at least one of;

a compound for reducing inflammation, and a compound for improving muscle fiber function, integrity and/or survival, and a pharmaceutically acceptable carrier, adjuvant, diluent and/or excipient.

39. A kit comprising a first compound for providing an individual with a functional dystrophin protein, and a second compound, wherein said second compound comprises at least one of:

a compound for reducing inflammation, and a compound for improving muscle fiber function, integrity and/or survival.

40. The kit of claim 39, further comprising a pharmaceutically acceptable carrier, adjuvant, diluent and/or excipient.

41. The kit of claim 39, further comprising packaging means thereof.