COMPOSITIONS AND METHODS FOR INHIBITING EXPRESSION OF HUNTINGTIN GENE

• US 20120095076A1
• Filed: 10/04/2011
• Published: 04/19/2012
• Est. Priority Date: 10/28/2005
• Status: Active Grant

• Current Assignee: Medtronic Incorporated (Medtronic Plc)
• Original Assignee: Alnylam Pharmaceuticals Incorporated
• Inventors: Sah, Dinah Wen-Yee, Hadwiger, Philipp, Roehl, Ingo, Bramlage, Birgit, Tan, Pamela, Vornlocher, Hans-Peter, Bumcrot, David
• Granted Patent: US 8,314,075 B2
• Time in Patent Office: Days
• US Class Current: 514/44.A
• 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 pharmaceutical composition comprising a double stranded ribonucleic acid (dsRNA) comprising a sense strand and an antisense strand and a pharmaceutically acceptable carrier, 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.

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

12. The dsRNA of claim 11, 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.

13. The dsRNA of claim 11, 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.

16. The pharmaceutical composition of claim 1, wherein said dsRNA comprises a cholesterol moiety.

17. A method for inhibiting expression of Huntingtin (HD) gene in a cell, the method comprising:

(a) introducing into the cell a the pharmaceutical composition of claim 1, 11, 12, 13, or 38; 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.

18. 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 1, 11, 12, or 13 or 38.

19. The method of claim 18, wherein the pharmaceutical composition is administered to the brain of the patient.

20. The method of claim 18, wherein the pharmaceutical composition is administered by intrastriatal infusion.

21. The method of claim 19, wherein administering the pharmaceutical composition to the brain causes a decrease in Huntingtin (HD) gene mRNA in the striatum.

22. The method of claim 19, wherein administering the pharmaceutical composition to the brain causes a decrease in Huntingtin (HD) gene mRNA in the cortex.

25. The method of claim 17, wherein the dsRNA of the pharmaceutical composition comprises a cholesterol moiety.

26. The method of claim 17, wherein the pharmaceutical composition is administered in vivo to a mammal.

27. The method of claim 25, wherein the pharmaceutical composition is administered in vivo to a mammal.

28. The method of claim 17, wherein the pharmaceutical composition is administered by intrastiatal infusion.

29. The method of claim 25, wherein the pharmaceutical composition is administered by intrastiatal infusion.

30. The method of claim 18, wherein the dsRNA of the pharmaceutical composition comprises a cholesterol moiety.

31. The method of claim 30, wherein the pharmaceutical composition is administered to the brain of the patient.

32. The method of claim 30, wherein the pharmaceutical composition is administered by intrastriatal infusion.

33. The method of claim 30, wherein administering the pharmaceutical composition to the brain causes a decrease in Huntingtin (HD) gene mRNA in the striatum.

34. The method of claim 30, wherein administering the pharmaceutical composition to the brain causes a decrease in Huntingtin (HD) gene mRNA in the cortex.

35. The pharmaceutical composition of claim 1, wherein the pharmaceutically acceptable carrier consists of an aqueous buffer.

36. The method of claim 17, wherein the pharmaceutically acceptable carrier consists of an aqueous buffer.

37. The method of claim 18, wherein the pharmaceutically acceptable carrier consists of an aqueous buffer.

38. 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.

39. The method of claim 17, 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.

40. The method of claim 18, 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.

2. (canceled)

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41. 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 N0:

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 N0;

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.

42. 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 N0:

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 N0;

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.