Cationic cyclic amine and amphipathic transfection reagents

Abstract

Cationic cyclic amine containing polymers and copolymers as well as novel lipids have been designed and synthesized for efficient delivery of nucleic acids to cells in biological systems, specifically for in vitro cell transfection research.

1 - 4 . (canceled) 
     
     
         5 . A transfection reagent comprising an amphipathic compound having the structure: 
       
         
           
           
               
               
           
         
         wherein 
         m is 2, 3, 4, 5, or 6; 
         n is 0 or 1; 
         R 1  and R 2  are each a lipid group; and 
         R 3  and R 4  are each selected from the group consisting of:
 (i) hydrogen, 
 (ii) —C(═O)—R 6 , 
 (iii) an amino acid linked by an amide bond, or 
 (iv) 2-pyridyl, 
 
       
       
         
           
           
               
               
           
         
         wherein 
         k is 0, 1, 2, 3, or 4; and 
         R 6  is 
       
       
         
           
           
               
               
           
         
       
     
     
         6 . The transfection reagent of  claim 5 , wherein said lipid group is a linear C 10 -C 20  alkyl, a branched C 10 -C 20  alkyl, a linear C 10 -C 20  alkenyl, a branched C 10 -C 20  alkenyl, or —C(═O)—R 5 ;
 wherein each R 5  is a linear C 9 -C 19  alkyl, a branched C 9 -C 19  alkyl, a linear C 9 -C 19  alkenyl, or a branched C 9 -C 19  alkenyl. 
 
     
     
         7 . The transfection reagent of  claim 6 , wherein each said lipid group is —C(═O)—R 5 . 
     
     
         8 . The transfection reagent of  claim 7 , wherein said R 5  is said linear C 9 -Cig alkenyl, or said branched C 9 -C 19  alkenyl, wherein each said linear C 9 -C 19  alkenyl or said branched C 9 -C 19  alkenyl comprises 1, 2, or 3 double bonds. 
     
     
         9 . The transfection reagent of  claim 6 , wherein each said lipid group is said linear C 10 -C 20  alkenyl, or said branched C 10 -C 20  alkenyl. 
     
     
         10 . The transfection reagent of  claim 9 , wherein each said linear C 10 -C 20  alkenyl or said branched C 10 -C 20  alkenyl comprises 1, 2, or 3 double bonds. 
     
     
         11 . The transfection reagent of  claim 10 , wherein each of said 1, 2, or 3 double bonds are cis-double bonds. 
     
     
         12 . The transfection reagent of  claim 5 , wherein said R 3  and said R 4  are each hydrogen. 
     
     
         13 . The transfection reagent of  claim 5 , wherein said R 3  and said R 4  are each —C(═O)—R 6 , wherein each said R 6  is 
       
         
           
           
               
               
           
         
       
     
     
         14 . The transfection reagent of  claim 5 , wherein said R 3  and said R 4  are each an amino acid linked by an amide bond. 
     
     
         15 . The transfection reagent of  claim 5 , wherein each said m is 3 or 4. 
     
     
         16 . The transfection reagent of  claim 15 , wherein said n is 0. 
     
     
         17 . The transfection reagent of  claim 16 , wherein said m is 4, said n is 0, said R 1  and said R 2  are each said linear C 10 -C 20  alkenyl, or said branched C 10 -C 20  alkenyl, and said R 3  and said R 4  are each hydrogen. 
     
     
         18 . The transfection reagent of  claim 17 , comprising an amphipathic compound having the structure: 
       
         
           
           
               
               
           
         
       
     
     
         19 . A transfection reagent comprising said amphipathic compound of  claim 5 , and a nucleic acid. 
     
     
         20 . A method of transfecting a nucleic acid into a cell, comprising mixing said amphipathic compound of  claim 5  with said nucleic acid to produce a mixed solution, and contacting said cell with said mixed solution, thereby transfecting said nucleic acid into said cell. 
     
     
         21 . A transfection reagent, comprising an amphipathic compound having the structure: 
       
         
           
           
               
               
           
         
       
     
     
         22 . A transfection reagent comprising said amphipathic compound of  claim 21 , and a nucleic acid. 
     
     
         23 . A method of transfecting a nucleic acid into a cell, comprising mixing said amphipathic compound of  claim 21  with said nucleic acid to produce a mixed solution, and contacting said cell with said mixed solution, thereby transfecting said nucleic acid into said cell.