COMPOSITIONS AND METHODS FOR INHIBITING EXPRESSION OF HUNTINGTIN GENE

• US 20080221055A1
• Filed: 11/26/2007
• Published: 09/11/2008
• Est. Priority Date: 10/28/2005
• Status: Active Grant

• Current Assignee: Medtronic Incorporated (Medtronic Plc)
• Original Assignee: Alnylam Pharmaceuticals Incorporated
• Inventors: Tan, Pamela, Bramlage, Birgit, Hadwiger, Philipp, Bumcrot, David, Vornlocher, Hans-Peter, Roehl, Ingo, Sah, Dinah Wen-Yee
• Granted Patent: US 7,749,978 B2
• Time in Patent Office: Days
• US Class Current: 514/44
• 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: 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 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3515 Lipophilic moiety, e.g. cholesterol
• CPC Class Codes: C12N 2310/3521 Methyl

Abstract

The invention relates to a double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of the Huntingtin gene (HD gene), comprising an antisense strand having a nucleotide sequence which is less than 25 nucleotides in length and which is substantially complementary to at least a part of the HD gene. The invention also relates to a pharmaceutical composition comprising the dsRNA together with a pharmaceutically acceptable carrier; methods for treating diseases caused by the expression of the HD gene, or a mutant form thereof, using the pharmaceutical composition; and methods for inhibiting the expression of the huntingtin gene in a cell.

Claims

1. A double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of a human Huntingtin gene in a cell, wherein said dsRNA is 21 nucleotides in length and comprises at least two sequences that are complementary to each other and wherein a sense strand comprises a first sequence and an antisense strand comprises a second sequence comprising a region of complementarity which is substantially complementary to at least a part of a mRNA encoding HD, and wherein said region of complementarity is at least 19 nucleotides in length and wherein said dsRNA, upon contact with a cell expressing said Huntingtin, inhibits expression of said Huntingtin gene by at least 20%.

3. The dsRNA of claim 1, wherein said dsRNA comprises at least one modified nucleotide.

5. The dsRNA of claim 3, wherein said modified nucleotide is chosen from the group of:

a 2′

-O-methyl modified nucleotide, a nucleotide comprising a 5′

-phosphorothioate group, and a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group.

6. The dsRNA of claim 3, wherein said modified nucleotide is chosen from the group of:

a 2′

-deoxy-2′

-fluoro modified nucleotide, a 2′

-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide, 2′

-amino-modified nucleotide, 2′

-alkyl-modified nucleotide, morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide.

10. A pharmaceutical composition for inhibiting the expression of the HD gene in an organism, comprising the dsRNA of claim 1 and a pharmaceutically acceptable carrier.

13. A method for inhibiting expression of Huntingtin gene in a cell, the method comprising:

(a) introducing into the cell a double-stranded ribonucleic acid (dsRNA) of claim 1, and

(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of the Huntingtin gene, thereby inhibiting expression of the Huntingtin gene in the cell.

14. A method of treating, preventing or managing Huntingtin disease comprising administering to a patient in need of such treatment, prevention or management a therapeutically or prophylactically effective amount of a dsRNA of claim 1.

17. The method of claim 14, wherein said dsRNA is administered to the brain of the patient.

18. The method of claim 14, wherein said dsRNA is administered by intrastriatal infusion.

19. The method of claim 17, wherein administering the dsRNA of claim 1 to the brain causes a decrease in HD mRNA in the striatum.

20. The method of claim 17, wherein administering the dsRNA of claim 1 to the brain causes a decrease in HD mRNA in the cortex.

21. The dsRNA of claim 1, wherein the dsRNA agent comprises a cholesterol moiety.

2. (canceled)

4. (canceled)

7. (canceled)

8. (canceled)

9. (canceled)

11. (canceled)

12. (canceled)

15. A vector for inhibiting the expression of Huntingtin gene in a cell, said vector comprising a regulatory sequence operably linked to a nucleotide sequence that encodes at least one strand of a dsRNA, wherein one of the strands of said dsRNA is substantially complementary to at least a part of a mRNA encoding Huntingtin and wherein said dsRNA is at least 21 nucleotides in length and wherein said dsRNA, upon contact with a cell expressing said Huntingtin, inhibits the expression of said Huntingtin gene by at least 20%.

16. A cell comprising the vector of claim 15.