Cationic graft-copolymer for non-viral gene delivery vector

Abstract

A cationic graft-copolymer for a non-viral gene delivery vector comprising a unit derived from a cationic derivative of a water-soluble linear polymers having a hydroxyl groups, namely, a cationic polysaccharide of the following formula (1) and the cationic derivative of polyvinylalcohol of the following formula (2) or the cationic derivative of the partial hydrolyzed polyvinylalcohol of the following formula (3) and a unit derived from a polymerizable olefin compound of the following formula (4) a process for preparing the same and a transfection reagent made therefrom.

1 . A non-viral gene delivery vector formed from an aqueous solution of a cationic graft-copolymer of a water-soluble linear backbone polymer having hydroxyl groups, for a non-viral gene delivery vector, comprising a unit derived from a cationic water-soluble linear polysaccharide of the following formula (1)
   [C 6 H 7 O 2 (OH) 3-a (OX) a ] x H 2 O  (1)   
     or a unit derived from a water-soluble linear polyvinylalcohol of the following formula (2) or a partial hydrolyzed alcohol of the following formula (3)
   —[CH 2 CH(OH) 1-b (OX) b ] n —  (2) 
   —[CH 2 CH(OH) 1-b-c (OX) b (OAc) c ] n —  (3), 
 wherein X is a —(CH 2 ) m R 1  organic radical where R 1  is a member of the class consisting of: 
 —NH 2  radical, 
 —N(CH 3 ) 2  radical, 
 —N(C 2 H 5 ) 2  radical, 
 —N + (C 2 H 5 ) 3  radical, 
 —N + (CH 2 ) 2 CH 2 CH(OH)CH 3  radical, 
 —N + (C 2 H 5 ) 2 CH 2 CH(OH)CH 3  radical, 
 —N + (C 2 H 5 ) 2 (C 2 H 5 )N(C 2 H 5 ) 2  radical, 
 —C 6 H 4 NH 2  radical, 
 —COC 6 H 4 NH 2  radical, 
 —COR 2  radical where R 2  is —CH 2 NH 2  or —C 6 H 4 NH 2 , and 
 —CH 2 CH(OH)CH 2 R 3  radical where R 3  is —NH 2 , —N(CH 3 ) 2 , —N(C 2 H 5 ) 2 , or —N + (C 2 H 5 ) 3  radical, 
 where m is a natural number of 1 to 3, a is a positive number having a value of 0<a<3, b is a positive number having a value of 0<b<1, x and n are natural numbers having a value of 5 or more, 1>b+c, and Ac is acetyl radical; and 
 a unit derived from a polymerize-able olefin compound of the following formula (4): 
 
     
       
         
         
             
             
         
       
       wherein R 4 , R 5  and R 6  are each selected from the group consisting of hydrogen and CH 3 , and R 7  is a member of the group consisting of: 
     
     
       
         
         
             
             
         
       
       where R 8  is a member of the class consisting of: hydrogen, C 1 -C 12  alkyl radicals, cyclohexyl radical, C 1 -C 4  hydroxyalkyl radicals, C 1 -C 8  aminoalkyl radicals, C 1 -C 8  dialkylaminoalkyl radicals, glycidyl radical, tetrahydrofuran radical, C 1 -C 4  lower alkyl-substituted tetrahydrofuran radical, benzyl radical, a (CH 2 CH 2 O) y CH 2 CH 2 OH radical where y is a positive integer from 1 to 10, and —N(R 9 ) 2 , where the two R 9 's which may be the same or different, are either hydrogen or a C 1 -C 4  alkyl radical; 
     
     
       
         
         
             
             
         
       
       wherein R 10  is a C 1 -C 8  alkyl radical, phenyl radical, tolyl radical, pyridine radical, pyrrolidone radical; and 
     
     
       
         
         
             
             
         
       
       where R 11  is NH 2 , NHCH 3 , N,N-dimethylamino radical, N,N-dimethylaminopropylamino radical, and morpholine radical. 
     
   
   
       2 . A process for preparing a non-viral gene delivery vector formed from an aqueous solution of a cationic graft-copolymer of a water-soluble linear backbone polymer having hydroxyl groups, for a non-viral gene delivery vector, which comprises reacting a cationic water-soluble linear polysaccharide of the following formula (1)
   [C 6 H 7 O 2 (OH) 3-a (OX) a ] x H 2 O  (1)   
     or a unit derived from a water-soluble linear polyvinylalcohol of the following formula (2) or a partial hydrolyzed alcohol of the following formula (3)
   —[CH 2 CH(OH) 1-b (OX) b ] n —  (2) 
   —[CH 2 CH(OH) 1-b-c (OX) b (OAc) c ] n —  (3), 
 wherein X is a —(CH 2 ) m R 1  organic radical where R 1  is a member of the class consisting of: 
 —NH 2  radical, 
 —N(CH 3 ) 2  radical, 
 —N(C 2 H 5 ) 2  radical, 
 —N + (C 2 H 5 ) 3  radical, 
 —N + (CH 2 ) 2 CH 2 CH(OH)CH 3  radical, 
 —N + (C 2 H 5 ) 2 CH 2 CH(OH)CH 3  radical, 
 —N + (C 2 H 5 ) 2 (C 2 H 5 )N(C 2 H 5 ) 2  radical, 
 —C 6 H 4 NH 2  radical, 
 —COC 6 H 4 NH 2  radical, 
 —COR 2  radical where R 2  is —CH 2 NH 2  or —C 6 H 4 NH 2 , and 
 —CH 2 CH(OH)CH 2 R 3  radical where R 3  is —NH 2 , —N(CH 3 ) 2 , —N(C 2 H 5 ) 2 , or —N + (C 2 H 5 ) 3  radical, 
 where m is a natural number of 1 to 3, a is a positive number having a value of 0<a<3, b is a positive number having a value of 0<b<1, x and n are natural numbers having a value of 5 or more, 1>b+c, and Ac is acetyl radical; with a polymerize-able olefin compound of the formula (4′): 
 
     
       
         
         
             
             
         
       
       wherein R 4 , R 5  and R 6  are each selected from the group consisting of hydrogen and CH 3 , and R 7  is a member of the group consisting of: 
     
     
       
         
         
             
             
         
       
     
     where R 8  is a member of the class consisting of hydrogen, C 1 -C 12  alkyl radicals, cyclohexyl radical, C 1 -C 4  hydroxyalkyl radicals, C 1 -C 8  aminoalkyl radicals, C 1 -C 8  dialkylaminoalkyl radicals, glycidyl radical, tetrahydrofuran radical, C 1 -C 4  lower alkyl-substituted tetrahydrofuran radical, benzyl radical, a (CH 2 CH 2 O) y CH 2 CH 2 OH radical where y is a positive integer from 1 to 10, and —N(R 9 ) 2  where the two R 9 's which may be the same or different, are either hydrogen or a C 1 -C 4  alkyl radical; 
     
       
         
         
             
             
         
       
     
     where R 10  is a C 1 -C 8  alkyl radical, phenyl radical, tolyl radical, pyridine radical, pyrrolidone radical; and 
     
       
         
         
             
             
         
       
     
     where R 11  is NH 2 , NHCH 3 , N,N-dimethylamino radical, N,N-dimethylaminopropylamino radical, and morpholine radical. 
   
   
       3 - 6 . (canceled)