METHODS AND MEANS FOR TREATING DNA REPEAT INSTABILITY ASSOCIATED GENETIC DISORDERS

• US 20100184833A1
• Filed: 08/10/2007
• Published: 07/22/2010
• Est. Priority Date: 08/11/2006
• Status: Abandoned Application

• Current Assignee: BioMarin Technologies B.V.
• Original Assignee: BioMarin Technologies B.V.
• Inventors: Wansink, Derick Gert, Platenburg, Gerard Johannes, De Kimpe, Josephus Johannes
• Application Number:
  US12/377,160
• Publication Number:
  US 20100184833A1
• Time in Patent Office: Days
• US Class Current: 514/44.R
• 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 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 15/11 DNA or RNA fragments; Modified forms thereof DNA or RNA not used in recombinant technology, C07H21/00; Non-coding nucleic acids having a biological activity
• 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 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers

Abstract

The current invention provides for methods and medicaments that apply oligonucleotide molecules complementary only to a repetitive sequence in a human gene transcript, for the manufacture of a medicament for the diagnosis, treatment or prevention of a cis-element repeat instability associated genetic disorders in humans. The invention hence provides a method of treatment for cis-element repeat instability associated genetic disorders. The invention also pertains to modified oligonucleotides which can be applied in method of the invention to prevent the accumulation and/or translation of repeat expanded transcripts in cells.

Claims

1. A method for preventing or treating a genetic disorder in a subject, comprising administering to a subject with a genetic disorder that is associated with human cis-element repeat instability a single stranded oligonucleotide comprising or consisting of a sequence that is complementary only to a repetitive element sequence in a gene transcript of a gene with said repeat instability.

2. The method according to claim 1 wherein the repetitive element is present in a coding sequence of the gene transcript.

3. The method according to claim 1 wherein the repetitive element is present in a non-coding sequence of the gene transcript.

4. The method according to claim 1 wherein the sequence of the repetitive element is selected from the group consisting of CAG;

GCG;

CUG;

CGG;

CCG;

GAA;

GCC; and

CCUG.

5. The method according to claim 2, wherein the oligonucleotide comprises or consists of a sequence that is complementary to a CAG repeat and wherein the disorder is Huntington'"'"'s disease, spino-cerebellar ataxias, Haw River syndrome, X-linked spinal and bulbar muscular atrophy or dentatorubral-pallidoluysian atrophy.

6. The method according to claim 2, wherein the oligonucleotide comprises or consists of a sequence that is complementary to a GCG repeat and wherein the disorder is infantile spasm syndrome, deidocranial dysplasia, blepharophimosis, hand-foot-genital disease, synpolydactyl, oculopharyngeal muscular dystrophy or holoprosencephaly.

7. The method according to claim 3, wherein the oligonucleotide comprises or consists of a sequence that is complementary to a CUG repeat and wherein the disorder is myotonic dystrophy type 1, spino-cerebellar ataxia 8 or Huntington'"'"'s disease-like 2.

8. The method according to claim 3 wherein the oligonucleotide comprises or consists of a sequence that is complementary to a CCUG repeat and wherein the disorder is myotonic dystrophy type 2.

9. The method according to claim 3, wherein the oligonucleotide comprises or consists of a sequence that is complementary to a CGG repeat and wherein the disorder is fragile X syndrome.

10. The method according to claim 3, wherein the oligonucleotide comprises or consists of a sequence that is complementary to a GAA repeat and wherein the disorder is Friedreich'"'"'s ataxia.

11. The method according to claim 1 wherein the oligonucleotide has a length of about 10 to about 50 nucleotides.

12. The method according to claim 1 wherein the single stranded oligonucleotide comprises ribonucleotides, deoxyribonucleotides, nucleotides of a locked nucleic acid (LNA), nucleotides of a peptide nucleic acid (PNA), morpholino phosphorodiamidates, nucleotides of an ethylene-bridged nucleic acid or a mixture thereof.

13. The method according to claim 12, wherein the oligonucleotide comprises 2′

-O-substituted RNA phosphorothioate nucleotides.

14. The method according to claim 1 wherein the administered oligonucleotide is in the form of an expressible nucleic acid vector.

15. The method according to claim 1, wherein the oligonucleotide is in a pharmaceutical composition which further comprises an excipient and/or targeting ligand that delivers the oligonucleotide to cells and/or enhances intracellular delivery of the oligonucleotide.

24. The method according to claim 11 wherein the oligonucleotide has a length of about 12 to about 30 nucleotides.

25. The method of claim 12 wherein the single stranded oligonucleotide has a phosphorothioate-containing backbone.

16. A single stranded oligonucleotide comprising or consisting of a sequence of about 10 to about 50 nucleotides that is complementary to a repetitive sequence of a tri- or tetranucleotide selected from the group consisting of

(a) CAG;

(b) GCG;

(c) CUG;

(c) CGG;

(d) GAA;

(e) GCC;

(f) CCUG.

17. The oligonucleotide according to claim 16, comprising 2′

-O-substituted phosphorothioate ribonucleotides, phosphorothioate deoxyribonucleotides, LNA nucleotides, morpholino nucleotides, and/or combinations thereof.

18. The oligonucleotide according to claim 26 wherein the label is a radioactive label or a fluorescent label.

19. A pharmaceutical composition comprising an oligonucleotide according to claim 16, and a pharmaceutically acceptable excipient.

20. The pharmaceutical composition of claim 19, further comprising a targeting ligand that delivers the oligonucleotide to a cell and/or enhances intracellular delivery of the oligonucleotide.

21. A nucleic acid vector, that expresses the oligonucleotide according to claim 16 in human cells.

22. A method for reducing the number of repeat-containing gene transcripts in a cell comprising providing to said cell the oligonucleotide according to claim 16.

26. The oligonucleotide of claim 16 that further comprises a detectable label.

27. A method for detecting the presence of nucleic acid repetitive elements in cells, comprising:

(a) contacting the cells or a lysate or extract thereof with the oligonucleotide according to claim 18, under conditions wherein said oligonucleotide hybridizes with cellular DNA and/or RNA;

(b) detecting hybridization of said oligonucleotide,

wherein the hybridization of said oligonucleotide is indicative of the presence of said repeat elements in said cells.

28. A method for diagnosing a genetic disorder associated with human cis-element repeat instability in a subject, comprising performing the method of claim 27 on cells, or obtained from a subject with, suspected of having, or at risk for said disorder, or on a lysate or extract of said cells,

wherein detection of the presence of said repeat elements in said cells, lysate or extract is diagnostic of said genetic disorder.

23. (canceled)