Treatment of genetic disorders associated with DNA repeat instability

• US 9,890,379 B2
• Filed: 07/27/2015
• Issued: 02/13/2018
• Est. Priority Date: 08/11/2006
• Status: Active Grant

• Current Assignee: Vico Therapeutics B.V.
• Original Assignee: BioMarin Technologies B.V.
• Inventors: De Kimpe, Josephus Johannes, Platenburg, Gerard Johannes, Wansink, Derick Gert
• Primary Examiner(s): ANGELL, JON E
• Application Number:
  US14/809,483
• Publication Number:
  US 20160053254A1
• Time in Patent Office: 932 Days
• 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 of treating in a subject having a genetic disorder associated with human cis-element repeat instability comprising administering to the subject an oligonucleotide comprising 10 to 50 nucleotides that is complementary to a sequence of a pre-mRNA transcript, said cis-element repeat instability is characterized by CAG repeat instability, said sequence comprising (CAGCAGCAGCAG) SEQ ID NO:

18, said oligonucleotide comprises either ribonucleotides or deoxyribonucleotides, said oligonucleotide comprising a modification selected from the group consisting of;

a morpholino phosphorodiamidate oligonucleotide, a phosphorothioate oligonucleotide, a locked nucleic acid (LNA), a peptide nucleic acid (PNA) and an ethylene-bridged nucleic acid, wherein said administered oligonucleotide does not induce RNaseH-mediated degradation of said pre-mRNA transcripts.

2. The method according to claim 1, wherein said sequence is present in a coding sequence of the gene pre-mRNA transcript.

3. The method according to claim 1, wherein said sequence is present in a non-coding sequence of the gene pre-mRNA transcript.

4. The method according to claim 1, wherein the disorder is Huntington'"'"'s disease.

5. The method according to claim 1, wherein the oligonucleotide comprises a 2′

-O-substituted phosphorothioate nucleotide.

6. The method of claim 5, wherein said oligonucleotide comprises a 2′

-O-methyl phosphorothioate nucleotide or a 2′

-O-methyoxy ethyl phosphorothioate nucleotide.

8. The method according to claim 1, wherein the oligonucleotide has a length selected from the group consisting of 12 to 30 nucleotides, 12 to 45 nucleotides and 12 to 25 nucleotides.

10. The method of claim 1, wherein the oligonucleotide is fully complementary to said sequence.

11. The method of claim 1, wherein the oligonucleotide is a 2′

-O-methyl phosphorothioate.

14. The method of claim 1, wherein the oligonucleotide is a RNA phosphorothioate comprising the sequence of SEQ ID NO:

5 (cug cug cug cug cug cug cug) or 20 (cug cug cug cug).

15. The method of claim 1, wherein the oligonucleotide is a DNA phosphorothioate oligonucleotide.

7. A method for reducing the number of CAG repeat-containing gene transcripts in a cell comprising providing to said cell an oligonucleotide comprising a 10 to 50 nucleotides that is complementary to a sequence of a pre-mRNA transcript, said sequence comprising (CAGCAGCAGCAG) SEQ ID NO:

18, said oligonucleotide comprises either ribonucleotides or deoxyribonucleotides, said oligonucleotide comprising a modification selected from the group consisting of;

a morpholino phosphorodiamidate oligonucleotide, a phosphorothioate oligonucleotide, a locked nucleic acid (LNA), a peptide nucleic acid (PNA) and an ethylene-bridged nucleic acid, wherein said oligonucleotide does not induce RNaseH-mediated degradation of said pre-mRNA transcripts.

9. The method of claim 7, wherein the oligonucleotide has a length selected from the group consisting of 12 to 30 nucleotides, 12 to 45 nucleotides and 12 to 25 nucleotides.

12. A method of treating a subject having a genetic disorder associated with human cis-element CAG repeat instability comprising administering to the subject an oligonucleotide comprising 10 to 50 nucleotides that is completely complementary to a sequence of a pre-mRNA transcript, wherein the oligonucleotide comprises a sequence selected from the group consisting of:

SEQ ID NOs;

5 (cug cug cug cug cug cug cug) and 20 (cug cug cug cug).

13. The method of claim 12, wherein the oligonucleotide comprises a modification selected from the group consisting of:

a morpholino phosphorodiamidate oligonucleotide, a phosphorothioate oligonucleotide, a locked nucleic acid (LNA), a peptide nucleic acid (PNA) and an ethylene-bridged nucleic acid.

16. The method of claim 12, wherein the oligonucleotide is RNA phosphorothioate consisting of the sequence of SEQ ID NO:

5 (cug cug cug cug cug cug cug) or 20 (cug cug cug cug).

17. The method of claim 12, wherein the oligonucleotide consists of the sequence of SEQ ID NOs:

5 (cug cug cug cug cug cug cug) or 20 (cug cug cug cug).