Highly packed polycationic ammonium, sulfonium and phosphonium lipids

Abstract

The present invention discloses highly packed polycationic ammonium, sulfonium and phosphonium lipid compounds useful for making lipid aggregates for delivery of macromolecules and other compounds into cells. They are especially useful for the DNA-dependent transformation of cells. Methods for their preparation and use as intracellular delivery agents are also disclosed.

1 . A compound or polycation having the formula: 
       
         
           
           
               
               
           
         
       
       or salt thereof where: 
       x is an integer ranging from 1 to about 20; 
       l is an integer ranging from 1 to about 6; 
       r and s, independently of one another, are 0 or 1, wherein when r is 1, the N bonded to R 1  and A 1  has a positive charge, and when s is 1, the N bonded to R 2  and A 2  has a positive charge; 
       R A  and R B , independently of one another, are selected from the group consisting of H, or an alkyl, hydroalkyl or thiol-substituted alkyl group having from 1 to 6 carbon atoms; 
       R1 and R2, independently of one another, are selected from the group consisting of alkyl groups having 1 to about 6 carbon atoms; and 
       A1 and A2, independently of other A1 and A2 groups, are selected from the group consisting of a —CH(D-L) 2  and a —C(D-L) 3  group wherein D is selected from the group consisting of —CO—, —CO 2 —, —O—C—O—, —CO—N—, —O—CO—N—, —O—, and —S—, and L is selected from the group consisting of:
 (a) a straight chain or branched alkyl, alkenyl, or alkynyl group having from 2 to about 22 carbon atoms wherein one or more non-neighboring —CH 2 — groups can be replaced with an O or S atom; 
 (b) a substituted straight chain or branched alkyl, alkenyl, or alkynyl group having from 2 to about 22 carbon atoms wherein the substituent is an aromatic, alicyclic heterocyclic or polycyclic ring and wherein one or more of the non-neighboring neighboring —CH 2 — groups of said alkyl, alkenyl or alkynyl group can be substituted with an O or S atom; and 
 (c) an aromatic, alicyclic, heterocyclic and a polycyclic ring moiety. 
 
     
     
         2 . A lipid aggregate which comprises one or more compounds of  claim 1 . 
     
     
         3 . A method for delivery of a macromolecule to a cell comprising the step of contacting said cell with a composition which comprises the compound of  claim 1  and said macromolecule. 
     
     
         4 . A compound or polycation having the formula: 
       
         
           
           
               
               
           
         
       
       or salt thereof where: 
       x is an integer ranging from 1 to about 20; 
       l is an integer ranging from 1 to about 6; 
       r and s, independently of one another, are 0 or 1, wherein when r is 1, the N bonded to R 1  and A 1  has a positive charge, and when s is 1, the N bonded to R 2  and A 2  has a positive charge; 
       R A  and R B , independently of one another, are selected from the group consisting of H, or an alkyl, hydroalkyl or thiol-substituted alkyl group having from 1 to 6 carbon atoms; 
       R1 and R2, independently of one another, are selected from the group consisting of alkyl groups having 1 to about 6 carbon atoms; and 
       A1 and A2, independently of other A1 and A2 groups, are selected from the group consisting of a B-L group wherein B is selected from the group consisting of —CO—, —CO 2 —, —O—C—O—, —CO—N—, —O—CO—N—, —O—CH 2 —, —S—CH 2 —, —CH 2 —S—, and —CH 2 — and L is selected from the group consisting of:
 (a) a straight chain or branched alkyl, alkenyl, or alkynyl group having from 2 to about 22 carbon atoms wherein one or more non-neighboring —CH 2 — groups can be replaced with an O or S atom; 
 (b) a substituted straight chain or branched alkyl, alkenyl, or alkynyl group having from 2 to about 22 carbon atoms wherein the substituent is an aromatic, alicyclic heterocyclic or polycyclic ring and wherein one or more of the non-neighboring neighboring —CH 2 — groups of said alkyl, alkenyl or alkynyl group can be substituted with an O or S atom; and 
 (c) an aromatic, alicyclic, heterocyclic and a polycyclic ring moiety. 
 
     
     
         5 . A method for delivery of a macromolecule to a cell comprising the step of contacting said cell with a composition which comprises the compound of  claim 4  and said macromolecule. 
     
     
         6 . A compound or polycation having the formula: 
       
         
           
           
               
               
           
         
       
       or salt thereof where: 
       x is an integer ranging from 1 to about 20; 
       l is an integer ranging from 1 to about 6; 
       r and s, independently of one another, are 0 or 1, wherein when r is 1, the N bonded to R 1  and A 1  has a positive charge, and when s is 1, the N bonded to R 2  and A 2  has a positive charge; 
       R A  and R B , independently of one another, are selected from the group consisting of H, or an alkyl, hydroalkyl or thiol-substituted alkyl group having from 1 to 6 carbon atoms; 
       R1 and R2, independently of one another, are selected from the group consisting of alkyl groups having 1 to about 6 carbon atoms; and 
       A1 and A2, independently of other A1 and A2 groups, are selected from the group consisting of a substituted straight chain or branched alkyl, alkenyl, or alkynyl group having from 2 to about 22 carbon atoms wherein the substituent is an aromatic, alicyclic heterocyclic or polycyclic ring and wherein one or more of the non-neighboring neighboring —CH 2 — groups of said alkyl, alkenyl or alkynyl group can be substituted with an O or S atom. 
     
     
         7 . A method for delivery of a macromolecule to a cell comprising the step of contacting said cell with a composition which comprises the compound of  claim 6  and said macromolecule. 
     
     
         8 . A compound or polycation having the formula: 
       
         
           
           
               
               
           
         
       
       or salt thereof where: 
       x is an integer ranging from 1 to about 20; 
       l is an integer ranging from 1 to about 6; 
       r and s, independently of one another, are 0 or 1, wherein when r is 1, the N bonded to R 1  and A 1  has a positive charge, and when s is 1, the N bonded to R 2  and A 2  has a positive charge; 
       R A  and R B , independently of one another, are selected from the group consisting of H, or an alkyl, hydroalkyl or thiol-substituted alkyl group having from 1 to 6 carbon atoms; 
       R1 and R2, independently of one another, are selected from the group consisting of alkyl groups having 1 to about 6 carbon atoms; and 
       A1 and A2, independently of other A1 and A2 groups, are selected from the group consisting of a straight chain or branched alkyl, alkenyl, or alkynyl group having from 2 to about 22 carbon atoms wherein one or more of the non-neighboring neighboring —CH 2 — groups of said alkyl, alkenyl or alkynyl group can be substituted with an O or S atom. 
     
     
         9 . A method for delivery of a macromolecule to a cell comprising the step of contacting said cell with a composition which comprises the compound of  claim 8  and said macromolecule. 
     
     
         10 . A compound or polycation having the formula: 
       
         
           
           
               
               
           
         
       
       or salt thereof where: 
       x is an integer ranging from 1 to about 20; 
       l is an integer ranging from 1 to about 6; 
       r and s, independently of one another, are 0 or 1, wherein when r is 1, the N bonded to R 1  and A 1  has a positive charge, and when s is 1, the N bonded to R 2  and A 2  has a positive charge; 
       R A  and R B , independently of one another, are selected from the group consisting of H, or an alkyl, hydroalkyl or thiol-substituted alkyl group having from 1 to 6 carbon atoms; 
       R1 and R2, independently of one another, are selected from the group consisting of alkyl groups having 1 to about 6 carbon atoms; and 
       A1 and A2, independently of other A1 and A2 groups, are selected from the group consisting of a straight chain or branched alkyl group substituted with one or two SH groups within about 3 carbon atoms of the bond between A1 or A2 and N. 
     
     
         11 . A method for delivery of a macromolecule to a cell comprising the step of contacting said cell with a composition which comprises the compound of  claim 10  and said macromolecule.