Novel lipids for transfection of nucleic acids

Abstract

Cationic lipid compositions are provided that are useful for efficient delivery of macromolecules, such as nucleic acids, into a wide variety of eukaryotic cell types. Methods for using the compositions also are provided.

1 . What is claimed is:
 A lipid having the formula I or II:   
       
         
           
           
               
               
           
         
         wherein in compounds of formula I: 
         X is N(R 4 R 5 R 6 ) or dialkylphosphatidyl, or X is 
       
       
         
           
           
               
               
           
         
         Y and Z independently are selected from the group consisting of alkoxy, alkanoyloxy, alkylamine, alkyl urethane and alkyl guanidine 
         R 1  and R 2  independently are selected from the group consisting of hydrogen, C 1 -C 6  alkyl, alkylamine, alkylaminoalcohol, spermiyl, spermidyl and carboxyspermiyl, 
         R 3  is H or C 1 -C 4  alkyl, 
         R 4 , R 5 , and R 6  independently are selected from the group consisting of hydrogen, alkyl, alkenyl, aryl and alkylaryl, provided that at least one of R 4 , R 5 , and R 6  is a long chain alkyl or alkenyl, 
         W is short chain alkyl or alkylamino, and 
         R 7  is a negative charge or short chain alkyl, 
         and wherein in compounds of formula II, X 1  is selected from the group consisting of (CH 2 ) n , (CHOH) n , —NH—, —O—, a polyether, an ester linkage —S—, CONH, NHCO, a polyamide —NHCONH—, and NHC(═NH)NH, 
         Y 1  is selected from the group consisting of (CH 2 ) n , a polyether, NHCO, —CO—, and a polyamide, 
         Y 2  is selected from the group consisting of (CH 2 ) n , —NH—, —O—, a polyether, an ester linkage, —S—, CONH, NHCO, a polyamide —NHCONH—, and NHC(═NH)NH, 
         Z 1  and Z 2  are independently selected from the group consisting of hydrogen, primary alkylamine, secondary alkylamine, tertiary alkyl amine, quaternary alkylamine, amino alcohol, alkyl polyamine, spermidine, spermine, carboxy spermine, guanidinium, pyridinium, pyrollidinium, piperidinyium amino acyl, peptidyl, and protein, 
         R 8 , R 9 , R 10 , and R 11  are independently absent or selected from the group consisting of hydrogen, straight chain C 1 -C 20  alkyl, branched alkyl, cyclalkyl, straight chain alkenyl, branched alkenyl, cyclo alkenyl, and optionally substituted aryl, wherein said optional substitution, when present, comprises at least one functional group selected from the group consisting of —OH, NH 2 , —COOH, ester, amide, alkylamino, —NHCONH—, and NHC(═NH)NH], and 
         n=1-12. 
       
     
     
         2 . The composition comprising a lipid according to  claim 1  and a macromolecule. 
     
     
         3 . The composition according to  claim 2  wherein said macromolecule is a nucleic acid. 
     
     
         4 . The composition according to  claim 3 , wherein said nucleic acid comprises a DNA molecule. 
     
     
         5 . The composition according to  claim 4 , wherein said DNA molecule is a double stranded DNA molecule. 
     
     
         6 . The composition according to  claim 5 , wherein said DNA molecule is a plasmid. 
     
     
         7 . The composition according to  claim 6 , wherein said plasmid encodes an RNA molecule that is self complementary and that forms a region of double stranded RNA. 
     
     
         8 . The composition according to  claim 3 , wherein said nucleic acid comprises an RNA molecule. 
     
     
         9 . The composition according to  claim 8 , wherein said RNA molecule is a double stranded RNA molecule. 
     
     
         10 . The composition according to  claim 9 , wherein said RNA molecule is an siRNA. 
     
     
         11 . A method of introducing a nucleic acid into a cell, comprising contacting a eukaryotic cell with a composition according to  claim 3 . 
     
     
         12 . A kit for transfecting a cell, comprising a lipid according to  claim 1 . 
     
     
         13 . A composition comprising a lipid according to  claim 1  and a peptide or protein. 
     
     
         14 . A method of introducing a peptide or protein into a cell, comprising contacting a eukaryotic cell with a composition according to  claim 13 . 
     
     
         15 . A kit for introducing a desired peptide or protein into a cell, comprising a lipid according to  claim 1 . 
     
     
         16 . A method of introducing a desired molecule into a tissue, comprising contacting said tissue with a composition comprising said desired molecule and a lipid according to  claim 1 . 
     
     
         17 . A method according to  claim 16 , wherein said desired molecule is a nucleic acid, a peptide, or a protein.