Sirna silencing of genes expressed in cancer

Abstract

The present invention provides nucleic acid-lipid particles comprising siRNA molecules that silence genes expressed in cancer (e.g., Eg5, EGFR or XIAP) and methods of using such nucleic acid-lipid particles to silence Eg5, EGFR or XIAP gene expression.

1 . A nucleic acid-lipid particle comprising:
 an siRNA molecule that silences expression a gene expressed in cancer, said gene selected from the group consisting of Eg5, EGFR, XIAP, and combinations thereof;   a cationic lipid;   a non-cationic lipid; and
 a conjugated lipid that inhibits aggregation of said particle. 
   
     
     
         2 . The nucleic acid-lipid particle of  claim 1 , wherein said nucleic acid-lipid particle comprises an siRNA molecule comprising at least one sequence set forth in any one of Tables 1-9. 
     
     
         3 . (canceled) 
     
     
         4 . (canceled) 
     
     
         5 . (canceled) 
     
     
         6 . The nucleic acid-lipid particle of  claim 1 , wherein said cationic lipid is a member selected from the group consisting of N,N-dioleyl-N,N-dimethylammonium chloride (DODAC), N,N-distearyl-N,N-dimethylammonium bromide (DDAB), N-(1-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP), N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA), and N,N-dimethyl-2,3-dioleyloxy)propylamine (DODMA), 1,2-DiLinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 1,2-Dilinolenyloxy-N,N-dimethylaminopropane (DLendMA), and a mixture thereof. 
     
     
         7 . The nucleic acid-lipid particle of  claim 1 , wherein said cationic lipid is DLinDMA. 
     
     
         8 . The nucleic acid-lipid particle of  claim 1 , wherein said non-cationic lipid is a member selected from the group consisting of dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), egg phosphatidylcholine (EPC), distearoylphosphatidylcholine (DSPC), palmitoyloleyolphosphatidylglycerol (POPG), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, palmitoyloleoyl-phosphatidylethanolamine (POPE), 1-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), cholesterol, and a mixture thereof. 
     
     
         9 . The nucleic acid-lipid particle of  claim 1 , wherein said non-cationic lipid is DSPC. 
     
     
         10 . The nucleic acid-lipid particle of  claim 1 , wherein the conjugated lipid that inhibits aggregation of particles is a polyethyleneglycol (PEG)-dialkyloxypropyl (PEG-DAA) conjugate selected from the group consisting of a PEG-dilauryloxypropyl (C 12 ), a PEG-dimyristyloxypropyl (C 14 ), a PEG-dipalmityloxypropyl (C 16 ), and a PEG-distearyloxypropyl (C 18 ). 
     
     
         11 . The nucleic acid-lipid particle of  claim 10 , wherein the PEG-DAA conjugate is a PEG-dimyristyloxypropyl (C 14 ). 
     
     
         12 . The nucleic acid-lipid particle of  claim 1 , wherein said cationic lipid comprises from about 30 mol % to about 50 mol % of the total lipid present in said particle. 
     
     
         13 . The nucleic acid-lipid particle of  claim 1 , wherein said non-cationic lipid comprises from about 5 mol % to about 90 mol % of the total lipid present in said particle. 
     
     
         14 . The nucleic acid-lipid particle of  claim 10 , wherein said PEG-DAA conjugate comprises from 1 mol % to about 20 mol % of the total lipid present in said particle. 
     
     
         15 . The nucleic acid-lipid particle of  claim 1 , further comprising cholesterol. 
     
     
         16 . The nucleic acid-lipid particle of  claim 15 , wherein the cholesterol comprises from about 10 mol % to about 60 mol % of the total lipid present in said particle. 
     
     
         17 . (canceled) 
     
     
         18 . (canceled) 
     
     
         19 . (canceled) 
     
     
         20 . The nucleic acid-lipid particle of  claim 1 , wherein the nucleic acid is fully encapsulated in said nucleic acid-lipid particle. 
     
     
         21 . The nucleic acid-lipid particle of  claim 1 , wherein said particle has a nucleic acid:lipid ratio of from about 0.01 to about 0.2. 
     
     
         22 . The nucleic acid-lipid particle of  claim 1 , wherein said particle has a median diameter of less than about 150 nm. 
     
     
         23 . A composition comprising the nucleic acid-lipid particle of  claim 2  and a pharmaceutically acceptable carrier. 
     
     
         24 . A method of introducing an siRNA that silences expression of a Eg5, EGFR or XIAP gene into a cell, said method comprising:
 contacting said cell with a nucleic acid-lipid particle comprising   a cationic lipid;   a non-cationic lipid;   a conjugated lipid that inhibits aggregation of particles; and   said siRNA.   
     
     
         25 .- 33 . (canceled)