METHODS AND MEANS FOR TREATING DNA REPEAT INSTABILITY ASSOCIATED GENETIC DISORDERS

• US 20110184050A1
• Filed: 08/06/2010
• Published: 07/28/2011
• Est. Priority Date: 02/08/2008
• Status: Active Grant

• Current Assignee: BioMarin Technologies B.V.
• Original Assignee: Prosensa Holding B.V.
• Inventors: De Kimpe, Josephus Johannes, Platenburg, Gerardus Johannes
• Granted Patent: US 8,263,760 B2
• Time in Patent Office: Days
• US Class Current: 514/44.R
• 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 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 15/113 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 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/331 Universal or degenerate base
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3521 Methyl

Abstract

The current invention provides for methods and medicaments that apply an oligonucleotide comprising aninosine and/or an uracile and/or a nucleotide containing a base able to form a wobble base pair, said oligonucleotide being preferably RNAse H substantially independent and being 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 a modified oligonucleotide which can be applied in a method of the invention to prevent the accumulation and/or translation of repeat expanded transcripts in cells.

Claims

1. Use of an oligonucleotide comprising an inosine and/or an uracile and/or a nucleotide containing a base able to form a wobble base pair, said oligonucleotide comprising or consisting of a sequence that is complementary only to a repetitive sequence in a gene transcript for the manufacture of a medicament for the treatment or prevention of human cis-element repeat instability associated genetic disorders.

2. A use according to claim 1, wherein the inosine and/or an uracile and/or a nucleotide containing a base able to form a wobble base pair containing oligonucleotide is a single stranded oligonucleotide.

3. A use according to claim 1 or 2, wherein the repetitive element is present in a coding or in a non-coding sequence of the gene transcript.

4. A use according to any of the preceding claims wherein the oligonucleotide comprises or consists of a sequence that is complementary to a repetitive element selected from (CAG)n, (GCG)n, (CUG)n, (CGG)n, (CCG)n, (GAA)n, (GCC)n and (CCUG)n.

5. A use according to any of the preceding claims wherein the oligonucleotide has a length of about 9 to about 50 nucleotides, preferably 12 to 40 nucleotides, more preferably 15 to 30.

6. A use according to any of the preceding claims, wherein the oligonucleotide comprises or consists of RNA nucleotides, DNA nucleotides, locked nucleic acid (LNA) nucleotides, peptide nucleic acid (PNA) nucleotides, morpholino phosphorodiamidates, ethylene-bridged nucleic acid (ENA) nucleotides or mixtures thereof with or without a phosphorothioate containing backbone.

7. A use according to claim 6, wherein the oligonucleotide comprises 2′

-O—

substituted RNA phosphorothioate nucleotides, preferably wherein the 2′

-O-substitution is a methoxy ethyl and/or methyl and/or 2′

O,4′

C methylene bridge (LNA) and/or 2′

O, 4′

C constrained ethylene (cEt) and/or 2′

O, 4′

C constrained methoxyethylene (cMOEt).

8. A use preferably according any of the preceding claims wherein the oligonucleotide comprises or consists of a sequence that is complementary only to a repetitive sequence in a gene transcript for the manufacture of a medicament for the treatment or prevention of human cis-element repeat instability associated genetic disorders and wherein the oligonucleotide is a RNAse H substantially independent oligonucleotide and preferably wherein the oligonucleotide has a length of 9 to 50 nucleotides, is substituted as defined in claim 6 in at least one of its 5′

or 3′

end and comprises less than 9, more preferably less than 6 consecutive deoxyriboses in the rest of its sequence.

9. A use according to claim 8, wherein the oligonucleotide comprises no deoxyribose and the backbone is fully modified as defined in claim 6.

10. A use according to any of the preceding claims wherein the oligonucleotide in the medicament is provided by a nucleic acid vector capable of conferring expression of the oligonucleotide and/or wherein the oligonucleotide in the medicament is provided with at least an excipient and/or targeting ligand for delivery of the oligonucleotide to cells and/or enhancing its intracellular delivery.

13. An oligonucleotide according to claim 11 or 12 and further as defined in any one of claims 1 to 10 and which is preferably provided with a radioactive label or fluorescent label.

11. An oligonucleotide comprising an inosine and/or an uracile and/or a nucleotide containing a base able to form a wobble base pair, wherein said oligonucleotide comprises or consists of a sequence that is complementary to a repetitive sequence selected from the group consisting of (CAG)n, (GCG)n, (CUG)n, (CGG)n, (GAA)n, (GCC)n and (CCUG)n, and has a length of 9 to 50 nucleotides.

12. An oligonucleotide preferably according to claim 11, comprising or consisting of a sequence that is complementary to a repetitive sequence selected from the group consisting of (CAG)n, (GCG)n, (CUG)n, (CGG)n, (GAA)n, (GCC)n and (CCUG)n, having a length of 9 to 50 nucleotides and wherein the oligonucleotide is a RNAse H substantially independent oligonucleotide.

14. A pharmaceutically acceptable composition comprising an oligonucleotide as defined in any of claims 11 to 13 and preferably further comprising at least one excipient and/or targeting ligand for delivery of the oligonucleotide to the cell and/or enhancing its intracellular delivery.

15. A nucleic acid vector, preferably a viral vector, capable of conferring expression of an oligonucleotide as defined in any of claims 11 to 13 in human cells.

16. An in vitro method for the reduction of repeat containing gene transcripts in a cell comprising the administration of an oligonucleotide as defined in any one of claims 11 to 13 or a pharmaceutically acceptable composition as defined in claim 14, or a nucleic acid vector as defined in claim 15.

17. In vitro or ex vivo detection and/or diagnostic method wherein the oligonucleotide according to any of claims 11 to 13 is used.