Modified oligonucleotides
One aspect of the present invention relates to double-stranded RNA (dsRNA) agent capable of inhibiting the expression of a target gene. Other aspects of the invention relate to pharmaceutical compositions comprising these dsRNA molecules suitable for therapeutic use, and methods of inhibiting the expression of a target gene by administering these dsRNA molecules, e.g., for the treatment of various disease conditions.
1 . A double-stranded RNA (dsRNA) molecule capable of inhibiting the expression of a target gene, comprising a sense strand and an antisense strand, each strand having 14 to 40 nucleotides, wherein the antisense strand has sufficient complementarity to the target sequence to mediate RNA interference, wherein the dsRNA molecule comprises only one hexopyranose nucleotide and
wherein said hexopyranose nucleotide is in the antisense strand at one of positions 6, 7, 9, 12, 16, 20 and 21, counting from the 5′-end of the antisense strand or said hexopyranose nucleotide is in the sense strand at one of positions 3, 12, and 21, counting from the 5′-end,
wherein the dsRNA comprises a 2′-fluoro or 2′-OMe nucleotide in position complimentary to the hexopyranose nucleotide, and
wherein the hexopyranose nucleotide is selected from the group consisting of altriol nucleotide, glucopyranose nucleotide, mannopyranose nucleotide, allopyranose nucleotide, and galactopyranose nucleotide.
2 . The double-stranded RNA molecule of claim 1 , wherein the antisense strand comprises the hexopyranose nucleotide.
3 . The double-stranded RNA molecule of claim 1 , wherein the antisense strand comprises a hexopyranose nucleotide at one of positions 6, 7 and 16.
4 . The double-stranded RNA molecule of claim 3 , wherein the antisense strand comprises a hexopyranose nucleotide at one of positions 6 and 7.
5 . The double-stranded RNA molecule of claim 4 , wherein the antisense strand comprises a hexopyranose nucleoside at position 7.
6 . The double-stranded RNA molecule of claim 1 , wherein the sense strand comprises the hexopyranose nucleotide.
7 . The double-stranded RNA molecule of claim 1 , wherein the sense strand comprises the hexopyranose nucleotide at one of positions 3 and 12.
8 . The double-stranded RNA molecule of claim 1 , wherein the sense strand comprises the hexopyranose nucleotide at position 3.
9 . The double-stranded RNA molecule of claim 1 , wherein sense strand comprises a 5′-vinylphosphonate (VP) group.
10 . The double-stranded RNA molecule of claim 1 , wherein the hexopyranose nucleotide is selected from the group consisting of altriol nucleotide, glucopyranose nucleotide, and mannopyranose nucleotide.
11 . The double-stranded RNA molecule of claim 10 , wherein the hexopyranose nucleotide is selected an altriol nucleotide.
12 . A pharmaceutical composition comprising the dsRNA agent according to claim 1 alone or in combination with a pharmaceutically acceptable carrier or excipient.
13 . A gene silencing kit containing the dsRNA molecule of claim 1 .
14 . A method for silencing a target gene in a cell, the method comprising a step of introducing the dsRNA molecule of claim 1 into the cell.
15 . The dsRNA molecule of claim 1 , wherein the dsRNA comprises a ligand.
16 . The dsRNA molecule of claim 1 , wherein the dsRNA molecule does not comprise nucleotides other than hexopyranose, 2′-fluoro and 2-OMe nucleotides.
17 . The dsRNA molecule of claim 1 , wherein the sense and antisense strands are independently 19-25 nucleotides in length.
18 . The double-stranded RNA molecule of claim 15 , wherein the ligand is an ASGPR ligand.
19 . The double-stranded RNA molecule of claim 18 , wherein the ASGPR ligand is: