Compositions and methods for the prevention and treatment of Huntington's disease

• US 20030109476A1
• Filed: 08/07/2002
• Published: 06/12/2003
• Est. Priority Date: 08/07/2001
• Status: Abandoned Application

• Current Assignee: University of Delaware
• Original Assignee: University of Delaware
• Inventors: Parekh-Olmedo, Hetal, Kmiec, Eric B.
• Application Number:
  US10/215,432
• Publication Number:
  US 20030109476A1
• Time in Patent Office: Days
• US Class Current: 514/44
• 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 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: 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 2310/315 Phosphorothioates
• 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 2310/3521 Methyl
• CPC Class Codes: C12N 2310/53 partially self-complementary or closed

Abstract

The present invention provides compositions and methods for the prevention and treatment of a neurodegenerative disease, specifically Huntington'"'"'s disease. In particular, the invention provides single-stranded, modified oligonucleotides for the targeted alteration of the genetic sequence of the Huntington'"'"'s disease gene, and mehods of treating or preventing Huntington'"'"'s disease using the same.

Claims

1. An oligonucleotide for targeted alteration of the genetic sequence of the Huntington'"'"'s disease gene, comprising a single-stranded oligonucleotide having a DNA domain, said DNA domain having at least one mismatch with respect to the genetic sequence of the Huntington'"'"'s disease gene to be altered, and further comprising chemical modifications of the oligonucleotide, said chemical modifications selected from the group consisting of an O-methyl modification, an LNA modification including LNA derivatives and analogs, two or more phosphorothioate linkages on one or more termini, and a combination of any two or more of these modifications.

2. The oligonucleotide according to claim 1, wherein said oligonucleotide comprises two or more phosphorothioate linkages on at least the 3′

terminus.

3. The oligonucleotide according to claim 1, wherein said oligonucleotide comprises one or more 2′

-O-methyl analogs.

4. The oligonucleotide according to claim 1, wherein said oligonucleotide comprises an LNA nucleotide, including an LNA derivative or analog.

5. The oligonucleotide according to claim 1, wherein said oligonucleotide comprises a combination of at least two modifications selected from the group of a phosphorothioate linkage, a 2′

-O-methyl analog, a locked nucleotide analog and a ribonucleotide.

6. The oligonucleotide according to claim 1, further comprising at least one unmodified ribonucleotide.

7. The oligonucleotide according to claim 2, wherein said oligonucleotide comprises two or more phosphorothioate linkages on both termini.

9. A method of targeted alteration of the genetic material of the Huntington'"'"'s disease gene, comprising the step of combining the genetic material of the Huntington'"'"'s disease gene with an oligonucleotide according to claim 1 or claim 8.

10. A method of targeted alteration of the genetic material of the Huntington'"'"'s disease gene, comprising the step of administering to a cell extract an oligonucleotide of claim 1 or claim 8.

11. A method of targeted alteration of the genetic material of the Huntington'"'"'s disease gene, comprising the step of administering to a cell an oligonucleotide of claim 1 or claim 8.

12. The method according to claim 11, wherein said genetic material of the Huntington'"'"'s disease gene is a non-transcribed DNA strand of a duplex DNA.

13. An altered genetic material of the Huntington'"'"'s disease gene obtained by the method of claim 10.

14. A cell comprising the altered genetic material of the Huntington'"'"'s disease gene of claim 13.

15. A method of treating Huntington'"'"'s disease, comprising the step of administering to a subject an effective amount of an oligonucleotide according to claim 1 or claim 8.

16. A method of prophylactically treating the severity of Huntington'"'"'s disease, comprising the step of administering to a subject an effective amount of an oligonucleotide according to claim 1 or claim 8.

17. A method of inhibiting the formation of Huntingtin comprising protein aggregates in cells, said protein aggregates being a characteristic of Huntington'"'"'s disease, comprising the step of administering to a subject an effective amount of an oligonucleotide according to claim 1 or claim 8.

18. A method of reducing Huntingtin comprising protein aggregates in cells, said protein aggregates being a characteristic of Huntington'"'"'s disease, comprising the step of administering to a subject an effective amount of an oligonucleotide according to claim 1 or claim 8.

8. An oligonucleotide for targeted alteration of the genetic sequence of the Huntington'"'"'s disease gene, comprising a chimeric RNA/DNA oligonucleotide, said oligonucleotide having at least one mismatch with respect to the genetic sequence of the Huntington'"'"'s disease gene to be altered.

19. A method of treating Huntington'"'"'s disease, comprising administering to a subject an effective amount of an oligonucleotide, wherein said oligonucleotide comprises a single-stranded oligonucleotide having a DNA domain, said DNA domain does or does not hybridize to the genetic sequence of the Huntington'"'"'s disease gene, and further comprises chemical modifications of the oligonucleotide, said chemical modifications being selected from the group consisting of an o-methyl modification, an LNA modification including LNA derivatives and analogs, one or more phosphorothioate linkages on one or more termini, and a combination of any two or more of these modifications.

23. The method according to any one of claims 19-22, wherein said oligonucleotide does not hybridize to the genetic sequence of the Huntington'"'"'s disease gene.

24. The method according to any one of claims 19-22, wherein said oligonucleotide does hybridize to the genetic sequence of the Huntington'"'"'s disease gene and wherein said DNA domain of said oligonucleotide has at least one mismatch with respect to the genetic sequence of the Huntington'"'"'s disease gene to be altered.

25. The method according to any one of claims 19-22, wherein said oligonucleotide comprises one or more phosphorothioate linkages on at least the 3′

terminus.

26. The method according to claim 25, wherein said oligonucleotide comprises one or more phosphorothioate linkage on both termini.

27. The method according to claim 25, wherein said oligonucleotide comprises all phosphorothioate linkages.

28. The method according to any one of claims 19-22, wherein said oligonucleotide comprises a 2′

-O-methyl analog.

29. The method according to any one of claims 19-22, wherein said oligonucleotide comprises a combination of at least two modifications selected from the group of a phosphorothioate linkage, a 2′

-O-methyl analog, a locked nucleotide analog and a ribonucleotide.

30. The method according to any one of claims 19-22, wherein said oligonucleotide comprises at least one unmodified ribonucleotide.

31. The method according to claim 23, wherein said oligonucleotide comprises at least one unmodified ribonucleotide.

32. The method according to claim 24, wherein said oligonucleotide comprises at least one unmodified ribonucleotide.

33. The method according to any one of claims 19-22, wherein said oligonucleotide is about 4 nucleotides to about 25 nucleotides in length.

34. The method according to claim 33, wherein said oligonucleotide is about 4 nucleotides to about 15 nucleotides in length.

35. The method according to claim 33, wherein said oligonucleotide is about 4 nucleotides to about 9 nucleotides in length.

20. A method of preventing Huntington'"'"'s disease, comprising the step of administering to a subject an effective amount of an oligonucleotide, wherein said oligonucleotide comprises a single-stranded oligonucleotide having a DNA domain, said DNA domain does or does not hybridize to the genetic sequence of the Huntington'"'"'s disease gene, and further comprises chemical modifications of the oligonucleotide, said chemical modifications being selected from the group consisting of an o-methyl modification, an LNA modification including LNA derivatives and analogs, one or more phosphorothioate linkages on one or more termini, and a combination of any two or more of these modifications.

21. A method of reducing Huntingtin comprising protein aggregates in cells, said protein aggregates being a characteristic of Huntington'"'"'s disease, comprising the step of administering to a subject an effective amount of an oligonucleotide, wherein said oligonucleotide comprises a single-stranded oligonucleotide having a DNA domain, said DNA domain does or does not hybridize to the genetic sequence of the Huntington'"'"'s disease gene, and further comprises chemical modifications of the oligonucleotide, said chemical modifications being selected from the group consisting of an o-methyl modification, an LNA modification including LNA derivatives and analogs, one or more phosphorothioate linkages on one or more termini, and a combination of any two or more of these modifications.

22. A method of inhibiting the formation of Huntingtin comprising protein aggregates in cells, said protein aggregates being a characteristic of Huntington'"'"'s disease, comprising the step of administering to a subject an effective amount of an oligonucleotide, wherein said oligonucleotide comprises a single-stranded oligonucleotide having a DNA domain, said DNA domain does or does not hybridize to the genetic sequence of the Huntington'"'"'s disease gene, and further comprises chemical modifications of the oligonucleotide, said chemical modifications being selected from the group consisting of an o-methyl modification, an LNA modification including LNA derivatives and analogs, one or more phosphorothioate linkages on a terminus, and a combination of any two or more of these modifications.