dsRNA For Treating Viral Infection
The invention relates to double-stranded ribonucleic acids (dsRNAs) targeting gene expression of phosphatidylinositol 4-kinase (PI4K), in particular human phosphatidylinositol 4-kinase, catalytic, beta polypeptide (PIK4CB) or human phosphatidylinositol 4-kinase, catalytic, alpha polypeptide (PIK4CA), and their use for treating infection by positive stranded RNA viruses such as hepatitis C virus (HCV). Each dsRNA comprises an antisense strand having a nucleotide sequence which is less that 30 nucleotides in length, generally 19-25 nucleotides in length, and which is substantially complementary to at least a part of the PIK4CB or PIK4CA target mRNA. A plurality of such dsRNA may be employed to provide therapeutic benefit. The invention also relates to a pharmaceutical composition comprising the dsRNA together with a pharmaceutically acceptable carrier, and including a delivery modality such as fully encapsulated liposomes or lipid complexes. The invention further includes methods for treating diseases caused by positive stranded RNA virus infection using the pharmaceutical compositions; and methods for inhibiting the propogation of positive stranded RNA viruses in and between cells.
1 . A double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of phosphatidylinositol 4-kinase (PI4K) in a cell, wherein said dsRNA comprises a first strand and a second strand, wherein the sequence of the first strand comprises the sequence of nucleotides 1 to 19 of SEQ ID NO:210.
2 . The dsRNA of claim 1 , wherein the sequence of the second strand comprises the sequence of nucleotides 1 to 19 of SEQ ID NO:314.
3 . The dsRNA of claim 1 , wherein said dsRNA comprises at least one modified nucleotide.
4 . The dsRNA of claim 3 , 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.
5 . The dsRNA of claim 3 , 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.
6 . A pharmaceutical composition comprising the dsRNA of claim 1 and a pharmaceutically acceptable carrier.
7 . A composition comprising the dsRNA of claim 1 , further comprising one or more additional dsRNAs for inhibiting the expression of phosphatidylinositol 4-kinase (PI4K) in a cell.
8 . A method for inhibiting the expression of the phosphatidylinositol 4-kinase, catalytic, beta polypeptide (PIK4CB) gene in a cell, the method comprising the steps of:
(a) introducing into the cell a double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of phosphatidylinositol 4-kinase (PI4K) in a cell, wherein said dsRNA comprises a first strand and a second strand, wherein the sequence of the first strand comprises the sequence of nucleotides 1 to 19 of SEQ ID NO:210; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of the PIK4CB gene, thereby inhibiting expression of the PIK4CB gene in the cell.
9 . The method of claim 8 , wherein the sequence of the second strand comprises the sequence of nucleotides 1 to 19 of SEQ ID NO: 314.
10 . The method of claim 8 , wherein said dsRNA comprises at least one modified nucleotide.
11 . The method of claim 8 , 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.
12 . The method of claim 8 , 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.
13 . A vector for inhibiting the expression of the phosphatidylinositol 4-kinase, catalytic, beta polypeptide (PIK4CB) gene in a cell, said 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 . A double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of phosphatidylinositol 4-kinase (PI4K) in a cell, wherein said dsRNA comprises a first strand and a second strand, wherein the sequence of the first strand comprises the sequence of SEQ ID NO:210 or SEQ ID NO:314.
16 . The dsRNA of claim 15 wherein said dsRNA comprises SEQ ID NO:210 and SEQ ID NO:314.