Compositions and methods directed to treating liver fibrosis

Abstract

The invention relates to double-stranded ribonucleic acid (dsRNA) compositions targeting the COL1A1, TGF-β, and SMAD2/3 genes, and methods of using such dsRNA compositions to inhibit expression of COL1A1, TGF-β, and SMAD2/3.

1 .- 4 . (canceled) 
     
     
         5 . A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of TGFβ, wherein said dsRNA comprises a sense strand and an antisense strand, the sense strand comprising one of SEQ ID NOs: 67, 71, 73, 87, 89, 107, 109, 111, 125 and 127, and the antisense strand comprising, respectively, one of SEQ ID NOs: 68, 72, 74, 88, 90, 108, 110, 112, 126 and 128. 
     
     
         6 .- 7 . (canceled) 
     
     
         8 . The dsRNA of  claim 5 , wherein said dsRNA comprises at least one modified nucleotide. 
     
     
         9 . The dsRNA of  claim 8 , wherein at least one of said modified nucleotides 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. 
     
     
         10 .- 11 . (canceled) 
     
     
         12 . The dsRNA of  claim 5 , wherein each strand is no more than 30 nucleotides in length. 
     
     
         13 . The dsRNA of  claim 5 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide. 
     
     
         14 . The dsRNA of  claim 5 , further comprising a ligand. 
     
     
         15 . The dsRNA of  claim 14 , wherein the ligand is conjugated to the 3′ end of the sense strand of the dsRNA. 
     
     
         16 .- 21 . (canceled) 
     
     
         22 . A cell containing the dsRNA of any of  claim 5 . 
     
     
         23 . (canceled) 
     
     
         24 . A pharmaceutical composition for inhibiting expression of a TGFβ gene comprising the dsRNA of  claim 5 . 
     
     
         25 . (canceled) 
     
     
         26 . A lipid formulation comprising a dsRNA of  claim 5 . 
     
     
         27 . The lipid formulation of  claim 26 , wherein the lipid formulation is a SNALP, or XTC formulation. 
     
     
         28 . A method of inhibiting TGFβ expression in a cell, the method comprising:
 a. introducing into the cell the dsRNA of  claim 5 ; and 
 b. maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a TGFβ gene, thereby inhibiting expression of the TGFβ gene in the cell. 
 
     
     
         29 . The method of  claim 28 , wherein the cell is a hepatic stellate cell. 
     
     
         30 . The method of  claim 28 , wherein the TGFβ expression is inhibited by at least 30%. 
     
     
         31 . A method of treating liver fibrosis comprising administering to a human in need of such treatment a therapeutically effective amount of the dsRNA of  claim 5 . 
     
     
         32 . The method of  claim 31 , wherein the dsRNA is administered at a concentration of 0.01 mg/kg-5 mg/kg bodyweight of the subject. 
     
     
         33 .- 34 . (canceled) 
     
     
         35 . A vector encoding a dsRNA, wherein said dsRNA comprises a sense strand and an antisense strand, the sense strand comprising one of SEQ ID NOs: 67, 71, 73, 87, 89, 107, 109, 111, 125 and 127, and the antisense strand comprising, respectively, one of SEQ ID NOs: 68, 72, 74, 88, 90, 108, 110, 112, 126 and 128. 
     
     
         36 .- 37 . (canceled) 
     
     
         38 . A cell comprising the vector of  claim 35 . 
     
     
         39 .- 40 . (canceled) 
     
     
         41 . The double-stranded ribonucleic acid of  claim 5 , wherein the sense strand comprises nucleotide sequence of SEQ ID NO: 71 and the antisense strand comprises nucleotide sequence of SEQ ID NO: 72. 
     
     
         42 . The double-stranded ribonucleic acid of  claim 5 , wherein the sense strand comprises nucleotide sequence of SEQ ID NO: 107 and the antisense strand comprises nucleotide sequence of SEQ ID NO: 108.