Compositions and Methods for Inhibiting Expression of Factor V
The invention relates to a double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of Factor V, comprising an antisense strand having a nucleotide sequence which is less that 25 nucleotides in length and which is substantially complementary to at least a part of Factor V. 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 Factor V using the pharmaceutical composition; and methods for inhibiting the expression of Factor V in a cell.
1 . An isolated double-stranded ribonucleic acid (dsRNA) comprising a sense strand and an antisense strand and wherein the sense strand consists of the nucleotide sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, or 75 and the antisense strand consists of the nucleotide sequence of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, or 76, respectively.
2 . The dsRNA of claim 1 , wherein the sense strand consists of the nucleotide sequence of SEQ ID NO: 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, or 75 and the antisense strand consists of the nucleotide sequence of SEQ ID NO: 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, or 76, respectively.
3 . The dsRNA of claim 1 , wherein the sense strand consists of the nucleotide sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, or 25 and the antisense strand consists of the nucleotide sequence of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, or 26, respectively.
4 . The dsRNA of claim 1 , wherein the dsRNA comprises at least one modified nucleotide.
5 . The dsRNA of claim 4 , 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.
6 . The dsRNA of claim 4 , wherein said modified nucleotide is chosen from the group of: 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.
7 . An isolated cell comprising the dsRNA of claim 1 .
8 . The cell of claim 7 , wherein the cell is a mammalian cell.
9 . A pharmaceutical composition comprising the dsRNA of claim 1 and a pharmaceutically acceptable carrier.
10 . A method for inhibiting the expression of a Factor V gene in a cell, the method comprising:
(a) introducing into the cell the dsRNA of claim 1 ; and
(b) maintaining the cell for a time sufficient to obtain degradation of the mRNA transcript of the Factor V gene, thereby inhibiting the expression of the Factor V gene in the cell.
11 . The cell of claim 10 , wherein the cell is a mammalian cell.
12 . A method of treating, preventing or managing thrombophilia comprising administering to a subject in need of such treatment, prevention or management a therapeutically or prophylactically effective amount of the dsRNA of claim 1 .
13 . A vector comprising a regulatory sequence operably linked to a nucleotide sequence that encodes at least one strand of the dsRNA of claim 1 .
14 . A cell comprising the vector of claim 13 .
15 . The cell of claim 14 , wherein the cell is a mammalian cell.