NOVEL COMPOSITIONS OF CHEMICALLY MODIFIED SMALL INTERFERING RNA
The present invention is directed to compositions comprising chemically modified siRNA that have high specificity by virtue of no or insignificant off-target activity of the sense strand, no or insignificant induction of IFN-like responses, high potency to offset oligonucleotide manufacturing costs, favorable manufacturing chemistry, and effective means of intracellular delivery both in vitro, during target validation and model studies, and in vivo, during animal model studies and clinical trials in humans.
1 . A DNA/RNA duplex comprising:
a) an antisense RNA strand of between 12 and 30 nucleotides;
b) a sense strand of between 12 and 30 nucleotides comprising:
i) a central region having between 3 and 10 2′-O-methylribonucleotides;
ii) two regions flanking and bound to said central region, each of said flanking
regions independently consisting of between 3 and 10 deoxyribonucleotides,
wherein said sense strand and said antisense strand are complementary to one another.
2 . The DNA/RNA duplex of claim 1 having the formula:
Sense
D D D D D O O O O O O O O O D D D D D
Antisense
R R R R R R R R R R R R R R R R R R R
wherein, each D is a deoxyribonucleotide; each O is a 2′-O-methylribonucleotide; and each R is a ribonucleotide.
3 . The DNA/RNA duplex of claim 2 , wherein the terminal 1 and 6 ribonculeotides at one or both ends of the antisense strand are bound to an adjacent ribonucleotide through a 2-3 phosphorothioate (PS) linkage.
4 . The DNA/RNA duplex of claim 2 , wherein between 1 and 5 deoxyribonucleotides in any flanking region of the sense strand are deoxyinosine.
5 . The DNA/RNA duplex of claim 1 having the formula:
Sense:
D I D I D O O O O O O O O O D I D I D
Antisense:
R′ R′ R′ R R R R R R R R R R R R R R′ R′ R′
wherein, each D is a deoxyribonucleotide; each O is a 2′-O-methylribonucleotide; each R is a ribonucleotide; each I is deoxyinosine; and each R′ is bound to an adjacent ribonucleotide through a 2-3 phosphorothioate (PS) linkage.