Compositions and methods for inhibiting expression of huntingtin gene
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.
1. A pharmaceutical composition comprising a double stranded ribonucleic acid (dsRNA) comprising a sense strand and an antisense strand and a pharmaceutically acceptable carrier, wherein the antisense strand comprises a nucleotide sequence consisting of SEQ ID NO:793 and a 2′-O-methyl modified nucleotide at location 4 and the sense strand comprises a nucleotide sequence consisting of SEQ ID NO:792 and a 2′-O-methyl modified nucleotide at locations 2, 3, 5, 12, 13, 15, and 17.
2. The dsRNA of claim 1 , wherein said dsRNA comprises at least one additional modified nucleotide.
3. The dsRNA of claim 2 , 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.
4. The dsRNA of claim 2 , wherein said modified nucleotide is chosen from the group of: a 5-bromo-2′-deoxyuridine, 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.
5. The pharmaceutical composition of claim 1 , wherein said dsRNA comprises a cholesterol moiety.
6. A method for inhibiting expression of Huntingtin (HD) gene in a cell, the method comprising: (a) introducing into the cell the pharmaceutical composition of claim 2 , 3 , or 4 ; and (b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of the HD gene, thereby inhibiting expression of the HD gene in the cell.
7. A method of treating or managing Huntingtin disease comprising administering to a patient in need of such treatment or management a therapeutically effective amount of the pharmaceutical composition of claim 2 , 3 , or 4 .
8. The method of claim 7 , wherein the pharmaceutical composition is administered to the brain of the patient.
9. The method of claim 7 , wherein the pharmaceutical composition is administered by intrastriatal infusion.
10. The method of claim 8 , wherein administering the pharmaceutical composition to the brain causes a decrease in Huntingtin (HD) gene mRNA in the striatum.
11. The method of claim 8 , wherein administering the pharmaceutical composition to the brain causes a decrease in Huntingtin (HD) gene mRNA in the cortex.
12. The method of claim 6 , wherein the dsRNA of the pharmaceutical composition comprises a cholesterol moiety.
13. The method of claim 6 , wherein the pharmaceutical composition is administered in vivo to a mammal.
14. The method of claim 12 , wherein the pharmaceutical composition is administered in vivo to a mammal.
15. The method of claim 6 , wherein the pharmaceutical composition is administered by intrastiatal infusion.
16. The method of claim 12 , wherein the pharmaceutical composition is administered by intrastiatal infusion.
17. The method of claim 7 , wherein the dsRNA of the pharmaceutical composition comprises a cholesterol moiety.
18. The method of claim 17 , wherein the pharmaceutical composition is administered to the brain of the patient.
19. The method of claim 17 , wherein the pharmaceutical composition is administered by intrastriatal infusion.
20. The method of claim 18 , wherein administering the pharmaceutical composition to the brain causes a decrease in Huntingtin (HD) gene mRNA in the striatum.
21. The method of claim 18 , wherein administering the pharmaceutical composition to the brain causes a decrease in Huntingtin (HD) gene mRNA in the cortex.
22. The pharmaceutical composition of claim 1 , wherein the pharmaceutically acceptable carrier consists of an aqueous buffer.
23. The method of claim 6 , wherein the pharmaceutically acceptable carrier consists of an aqueous buffer.
24. The method of claim 7 , wherein the pharmaceutically acceptable carrier consists of an aqueous buffer.
25. The pharmaceutical composition of claim 1 , wherein the antisense strand comprises a phosphorothioate at the internucleotide linkage between nucleotides 20 and 21 and the sense strand comprises a phosphorothioate at the internucleotide linkage between nucleotides 20 and 21.
26. The method of claim 6 , wherein the antisense strand comprises a phosphorothioate at the internucleotide linkage between nucleotides 20 and 21 and the sense strand comprises a phosphorothioate at the internucleotide linkage between nucleotides 20 and 21.
27. The method of claim 7 , wherein the antisense strand comprises a phosphorothioate at the internucleotide linkage between nucleotides 20 and 21 and the sense strand comprises a phosphorothioate at the internucleotide linkage between nucleotides 20 and 21.
28. A method of inhibiting expression of Huntingtin (HD) gene in a cell in a patient comprising administering to the brain of the patient by intrastriatal infusion a therapeutically effective amount of a pharmaceutical composition comprising a dsRNA and an aqueous buffer, the dsRNA consisting of a sense strand and antisense strand, wherein the antisense strand comprises a nucleotide sequence consisting of SEQ ID NO:793 and a 2′-O-methyl modified nucleotide at location 4 and a phosphorothioate at the internucleotide linkage between nucleotides 20 and 21 and the sense strand comprises a nucleotide sequence consisting of SEQ ID NO:792 and said sense strand comprises a 2′-O-methyl modified nucleotide at locations 2, 3, 5, 12, 13, 15, and 17 and a phosphorothioate at the internucleotide linkage between nucleotides 20 and 21.
29. A method of treating or managing Huntintin disease in a patient comprising administering to the brain of the patient by intrastriatal infusion a therapeutically effective amount of a pharmaceutical composition comprising a dsRNA and an aqueous buffer, the dsRNA consisting of a sense strand and antisense strand, wherein the antisense strand comprises a nucleotide sequence consisting of SEQ ID NO:793 and a 2′-O-methyl modified nucleotide at location 4 and a phosphorothioate at the internucleotide linkage between nucleotides 20 and 21 and the sense strand comprises a nucleotide sequence consisting of SEQ ID NO:792 and a 2′-O-methyl modified nucleotide at locations 2, 3, 5, 12, 13, 15, and 17 and a phosphorothioate at the internucleotide linkage between nucleotides 20 and 21.