RNA interference mediated inhibition of checkpoint kinase-1 (CHK-1) gene expression using short interfering nucleic acid (siNA)
This invention relates to compounds, compositions, and methods useful for modulating checkpoint kinase (e.g., checkpoint kinase-1 or CHK-1) gene expression using short interfering nucleic acid (siNA) molecules. This invention also relates to compounds, compositions, and methods useful for modulating the expression and activity of other genes involved in pathways of checkpoint kinase gene expression and/or activity by RNA interference (RNAi) using small nucleic acid 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 checkpoint kinase genes.
1. A chemically modified short interfering nucleic acid molecule (siNA), wherein:
(a) the siNA molecule comprises a sense strand and a separate antisense strand, each strand having one or more pyrimidine nucleotides and one or more purine nucleotides;
(b) each strand is independently 18 to 27 nucleotides in length; and together comprise a duplex having between 17 and 23 base pairs;
(c) the antisense strand is complementary to a human checkpoint kinase-1 (CHK-1) RNA sequence comprising SEQ ID NO:277;
(d) a plurality of the pyrimidine nucleotides present in the sense strand are 2′-deoxy-2′-fluoro pyrimidine nucleotides and a plurality of the purine nucleotides present in the sense strand are 2′-deoxy purine nucleotides; and,
(e) plurality of the pyrimidine nucleotides in the antisense strand are 2′-deoxy-2′-fluoro pyrimidine nucleotides and a plurality of the purine nucleotides present in the antisense strand are 2′-O-methyl purine nucleotides.
2. The siNA molecule of claim 1 , wherein the sense strand includes a terminal cap moiety at both 5′- and 3′-ends.
3. The siNA molecule of claim 1 , wherein the sense strand, the antisense strand, or both the sense strand and the antisense strand comprise a 3′-overhang.
4. A composition comprising the siNA molecule of claim 1 and a pharmaceutically acceptable carrier or diluent.
5. The siNA of claim 1 , wherein the antisense strand has a phosphorothioate internucleotide linkage at the 3′-end.
6. A chemically modified short interfering nucleic acid (siNA) molecule, wherein:
(a) the siNA molecule comprises a sense strand and a separate antisense strand, each strand having one or more pyrimidine nucleotides and one or more purine nucleotides;
(b) each strand is independently 18 to 27 nucleotides in length, and together comprise a duplex having between 17 and 23 base pairs;
(c) the antisense strand is complementary to a human checkpoint kinase-1 (CHK-1) RNA sequence comprising SEQ ID NO:277;
(d) a plurality of the pyrimidine nucleotides present in the sense strand are 2′-deoxy-2′-fluoro pyrimidine nucleotides and a plurality of the purine nucleotides present in the sense strand are 2′-deoxy purine nucleotides; and,
(e) a plurality of the pyrimidine nucleotides present in the antisense strand are 2′-deoxy-2′-fluoro pyrimidine nucleotides and a plurality of the purine nucleotides present in the antisense strand are 2′-deoxy purine nucleotides.
7. The siNA molecule of claim 6 , wherein the antisense strand has a phosphorothioate internucleotide linkage at the 3′-end.
8. The siNA molecule of claim 6 , wherein the sense strand includes a terminal cap moiety at both 5′- and 3′- ends.
9. The siNA molecule of claim 6 , wherein the sense strand, the antisense strand, or both the sense strand and the antisense strand comprise a 3′-overhang.
10. A composition comprising the siNA molecule of claim 6 and a pharmaceutically acceptable carrier or diluent.