dsRNA For Treating Viral Infection

Abstract

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 propagation of positive stranded RNA viruses in and between cells.

1 .- 63 . (canceled) 
   
   
       64 . A double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of phosphatidylinositol 4-kinase (PI4K) in a cell, wherein said dsRNA comprises a sense strand and an anti-sense strand that are complementary to each other, wherein the sense strand comprises a first sequence;
 and wherein the antisense strand comprises a second sequence comprising a region of complementarity which is substantially complementary to at least a part of a mRNA encoding PI4K,   wherein said region of complementarity is less than 30 nucleotides h length;   and wherein said dsRNA, upon contact with a cell expressing said PI4K gene, inhibits expression of said PI4K gene.   
   
   
       65 . The dsRNA of  claim 64 , wherein said second sequence comprises a sequence which is substantially complementary to at least part of an mRNA encoding human phosphatidylinositol 4-kinase, catalytic, beta polypeptide (PIK4CB). 
   
   
       66 . The dsRNA of  claim 65 , wherein said first sequence and/or said second sequence is selected from among the group consisting of Table 1. 
   
   
       67 . The dsRNA of  claim 64 , wherein said antisense strand comprises a sequence which is substantially complementary to at least part of an mRNA encoding human phosphatidylinositol 4-kinase, catalytic, alpha polypeptide (PIK4CA). 
   
   
       68 . The dsRNA of  claim 67 , wherein said first sequence and/or said second sequence is selected from among the group consisting of Table 2. 
   
   
       69 . The dsRNA of  claim 64 , wherein said dsRNA comprises at least one modified nucleotide. 
   
   
       70 . The dsRNA of  claims 69 , wherein said modified nucleotide is chosen from the group consisting of: a 2′-O-methyl modified nucleotide; a nucleotide comprising a 5′-phosphorothioate group; a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group; 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. 
   
   
       71 . The dsRNA of  claim 64 , wherein said contact reduces the expression level of PIK4CB by at least 40%. 
   
   
       72 . The dsRNA of  claim 64 , wherein said contact is performed in vitro at 30 nM or less. 
   
   
       73 . A cell comprising the dsRNA of  claim 64 . 
   
   
       74 . A pharmaceutical composition comprising the dsRNA of  claim 64  and a pharmaceutically acceptable carrier. 
   
   
       75 . The pharmaceutical composition of  claim 74 , wherein said dsRNA comprises at least one modified nucleotide. 
   
   
       76 . The pharmaceutical composition of  claim 74 , wherein the composition comprises a fully encapsulated liposome, a lipid complex, and/or a polymer. 
   
   
       77 . A method for inhibiting the expression of the phosphatidylinositol 4-kinase, catalytic, beta polypeptide (PIK4CB) gene in a cell, the method comprising:
 (a) introducing into the cell a dsRNA of  claim 64 ; 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.   
   
   
       78 . A method of treating a pathological processes mediated by positive stranded RNA virus infection comprising the steps of:
 providing a dsRNA according to  claim 64 ; and   administering to a patient in need of such treatment, prevention or management a therapeutically or prophylactically effective amount of the dsRNA.   
   
   
       79 . The method of  claim 78 , wherein said positive stranded RNA virus is selected from among hepatitis C virus (HCV), human papilloma virus (HPV), and Dengue virus. 
   
   
       80 . A double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of phosphatidylinositol 4-kinase (PI4K) in a cell, wherein said dsRNA comprises a sense strand and an anti-sense strand that are complementary to each other, wherein the sense strand comprises a first sequence;
 and wherein the antisense strand comprises a second sequence comprising a region of complementarity which is substantially complementary to at least a part of a mRNA encoding PI4K,   wherein said region of complementarity is less than 30 nucleotides in length;   and wherein said dsRNA, upon contact with a cell expressing said PI4K gene, inhibits expression of said PI4K gene;   and wherein said first sequence and/or said second sequence is selected from among the group consisting of Table 1.   
   
   
       81 . The dsRNA of  claim 80 , wherein said dsRNA comprises at least one modified nucleotide. 
   
   
       82 . The dsRNA of  claim 81 , wherein said modified nucleotide is chosen from the group consisting of: a 2′-O-methyl modified nucleotide; a nucleotide comprising a 5′-phosphorothioate group; a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group; 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. 
   
   
       83 . A double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of phosphatidylinositol 4-kinase (PI4K) in a cell, wherein said dsRNA comprises a sense strand and an anti-sense strand that are complementary to each other, wherein the sense strand comprises a first sequence;
 and wherein the antisense strand comprises a second sequence comprising a region of complementarity which is substantially complementary to at least a part of a mRNA encoding PI4K,   wherein said region of complementarity is less than 30 nucleotides in length;   and wherein said dsRNA, upon contact with a cell expressing said PI4K gene, inhibits expression of said PI4K gene;   and wherein said first sequence and/or said second sequence is selected from among the group consisting of Table 2.   
   
   
       84 . The dsRNA of  claim 83 , wherein said dsRNA comprises at least one modified nucleotide. 
   
   
       85 . The dsRNA of  claims 84 , wherein said modified nucleotide is chosen from the group consisting of: a 2′-O-methyl modified nucleotide; a nucleotide comprising a 5′-phosphorothioate group; a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group; 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.