RNA interference mediated inhibition of gene expression using short interfering nucleic acid (siNA)

• US 20070042983A1
• Filed: 08/20/2004
• Published: 02/22/2007
• Est. Priority Date: 05/18/2001
• Status: Abandoned Application

• Current Assignee: Sirna Therapeutics, Inc. (Alnylam Pharmaceuticals Incorporated)
• Original Assignee: Sirna Therapeutics, Inc. (Alnylam Pharmaceuticals Incorporated)
• Inventors: McSwiggen, James, Haeberli, Peter
• Application Number:
  US10/923,536
• Publication Number:
  US 20070042983A1
• Time in Patent Office: Days
• US Class Current: 514/44.A
• 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: C07H 21/02 with ribosyl as saccharide radical
• CPC Class Codes: C12P 19/34 Polynucleotides, e.g. nucleic acids, oligoribonucleotides

Abstract

This invention relates to compounds, compositions, and methods useful for modulating gene expression using short interfering nucleic acid (siNA) molecules. In particular, the instant invention features small nucleic acid molecules, such as short interfering nucleic acid (siNA), short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), and short hairpin RNA (shRNA) molecules and methods used to modulate the expression of genes, such as expressed pseudogenes associated with the maintenance or development of diseases, disorders, traits, and conditions in a subject or organism.

Claims

1-32. -32. (canceled)

33. A chemically synthesized double stranded short interfering nucleic acid (siNA) molecule, wherein:

a. the siNA comprises a first strand and a second strand having at least one chemical modification;

b. the first strand comprises a sense region the second strand comprises an antisense region; and

c. the first strand and second strand form an asymmetric duplex wherein the antisense region is of length from about 15 to about 30 nucleotides, and the sense region is of length from about 10 to about 25 nucleotides that are complementary to the antisense region.

34. The siNA molecule of claim 33, wherein said antisense region comprises a 5′

-terminal phosphate group.

35. The siNA molecule of claim 33, wherein said siNA molecule comprises no ribonucleotides.

36. The siNA molecule of claim 33, wherein said siNA molecule comprises one or more ribonucleotides.

37. The siNA molecule of claim 33, wherein the first strand includes a terminal cap moiety at the 5′

-end, the 3′

-end, or both of the 5′ and

3′

ends.

38. The siNA molecule of claim 33, wherein the second strand includes a terminal cap moiety at the 3′

-end.

39. The siNA molecule of claim 37, wherein said cap moiety comprises an abasic moiety.

40. The siNA molecule of claim 38, wherein said cap moiety comprises an abasic moiety.

41. The siNA molecule of claim 39, wherein said abasic moiety comprises an inverted deoxyabasic moiety.

42. The siNA molecule of claim 40, wherein said abasic moiety comprises an inverted deoxyabasic moiety.

43. The siNA molecule of claim 33, wherein pyrimidine nucleotides in the sense region are 2′

-O-methylpyrimidine nucleotides.

44. The siNA molecule of claim 33, wherein purine nucleotides in the sense region are 2′

-deoxy purine nucleotides.

45. The siNA molecule of claim 33, wherein pyrimidine nucleotides present in the sense region are 2′

-deoxy-2′

-fluoro pyrimidine nucleotides.

46. The siNA molecule of claim 33, wherein pyrimidine nucleotides of said antisense region are 2′

-deoxy-2′

-fluoro pyrimidine nucleotides

47. The siNA molecule of claim 33, wherein purine nucleotides of said antisense region are 2′

-O-methyl purine nucleotides.

48. The siNA molecule of claim 33, wherein said chemical modification comprises a modified nucleotide having a Northern configuration.

49. The siNA molecule of claim 48, wherein said Northern conformation nucleotides are selected from the group consisting of locked nucleic acid (LNA) nucleotides;

2′

-methoxyethoxy nucleotides;

2′

-methyl-thio-ethyl nucleotides, 2′

-deoxy-2′

-fluoro nucleotides, 2′

-deoxy-2′

-chloro nucleotides, 2-azido nucleotides, 2′

-O-trifluoromethyl nucleotides, 2′

-O-ethyl-trifluoromethoxy nucleotides, 2′

-O-difluoromethoxy-ethoxy nucleotides, 4′

-thio nucleotides and 2′

-O-methyl nucleotides.

50. A pharmaceutical composition comprising the siNA molecule of claim 33 in an acceptable carrier or diluent.