Lipid formulated dsrna targeting the pcsk9 gene

Abstract

This invention relates to composition and methods using lipid formulated siRNA targeted to a PCSK9 gene.

1 . A composition comprising a nucleic acid lipid particle comprising a double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of a human PCSK9 gene in a cell, wherein:
 the nucleic acid lipid particle comprises a lipid formulation comprising 45-65 mol % of a cationic lipid, 5 mol % to about 10 mol %, of a non-cationic lipid, 25-40 mol % of a sterol, and 0.5-5 mol % of a PEG or PEG-modified lipid,   the dsRNA consists of a sense strand and an antisense strand, and the sense strand comprises a first sequence and the antisense strand comprises a second sequence complementary to at least 15 contiguous nucleotides of a nucleotide sequence of a target sequence of a dsRNA found in Table 1a, Table 2a, Table 5a, Table 6, Table 7, Table 8,   wherein the first sequence is complementary to the second sequence and wherein the dsRNA is between 15 and 30 base pairs in length.   
     
     
         2 . The composition of  claim 1 , wherein the cationic lipid comprises MC3 (((6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate). 
     
     
         3 . The composition of  claim 2 , wherein the cationic lipid comprises MC3 and the lipid formulation is selected from the group consisting of: 
       
         
           
                 
                 
                 
               
                     
                 
                     
                   LNP11 
                   MC3/DSPC/Cholesterol/PEG-DMG 
                 
                     
                     
                   50/10/38.5/1.5 
                 
                     
                   LNP14 
                   MC3/DSPC/Cholesterol/PEG-DMG 
                 
                     
                     
                   40/15/40/5 
                 
                     
                   LNP15 
                   MC3/DSPC/Cholesterol/PEG-DSG/GalNAc-PEG-DSG 
                 
                     
                     
                   50/10/35/4.5/0.5 
                 
                     
                   LNP16 
                   MC3/DSPC/Cholesterol/PEG- DMG   
                 
                     
                     
                   50/10/38.5/1.5 
                 
                     
                   LNP17 
                   MC3/DSPC/Cholesterol/PEG-DSG 
                 
                     
                     
                   50/10/38.5/1.5 
                 
                     
                   LNP18 
                   MC3/DSPC/Cholesterol/PEG-DMG 
                 
                     
                     
                   50/10/38.5/1.5 
                 
                     
                   LNP19 
                   MC3/DSPC/Cholesterol/PEG-DMG 
                 
                     
                     
                   50/10/35/5 
                 
                     
                   LNP20 
                   MC3/DSPC/Cholesterol/PEG-DPG 
                 
                     
                     
                   50/10/38.5/1.5 
                 
                     
                 
             
                
               
               
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         4 .- 9 . (canceled) 
     
     
         10 . The composition of  claim 1 , wherein the sense strand comprises SEQ ID NO:1227 and the antisense strand comprises SEQ ID NO:1228. 
     
     
         11 . (canceled) 
     
     
         12 . (canceled) 
     
     
         13 . The composition of  claim 1 , wherein the dsRNA comprises at least one modified nucleotide. 
     
     
         14 . The composition of  claim 13 , wherein the 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. 
     
     
         15 . The composition of  claim 13 , wherein the 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. 
     
     
         16 . The composition of  claim 1 , wherein the dsRNA comprises at least one 2′-O-methyl modified ribonucleotide and at least one nucleotide comprising a 5′-phosphorothioate group. 
     
     
         17 . The composition of  claim 1 , wherein each strand of the dsRNA is 19-23 bases in length. 
     
     
         18 . The composition of  claim 1 , wherein each strand of the dsRNA is 21-23 bases in length. 
     
     
         19 . The composition of  claim 1 , wherein each strand of the dsRNA is 21 bases in length. 
     
     
         20 . The composition of  claim 1 , further comprising a lipoprotein. 
     
     
         21 . The composition of  claim 1 , further comprising apolipoprotein E (ApoE). 
     
     
         22 . The composition of  claim 21 , wherein the dsRNA is conjugated to a lipophile. 
     
     
         23 . The composition of  claim 22 , wherein the lipophile is cholesterol. 
     
     
         24 . The composition of  claim 21 , wherein the ApoE is reconstituted with at least one amphiphilic agent. 
     
     
         25 . The composition of  claim 24 , wherein the amphiphilic agent is a phospholipid. 
     
     
         26 . The composition of  claim 24 , wherein the amphilic agent is a phospholipid selected from the group consisting of dimyristoyl phosphatidyl choline (DMPC), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), egg phosphatidylcholine (EPC), di stearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), -phosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), and combinations thereof. 
     
     
         27 . The composition of  claim 25 , wherein the ApoE is reconstituted high density lipoprotein (rHDL). 
     
     
         28 .- 33 . (canceled) 
     
     
         34 . A method for inhibiting the expression of PCSK9 in a cell comprising administering the composition of  claim 1  to the cell. 
     
     
         35 .- 43 . (canceled)