Novel compositions for the delivery of negatively charged molecules

Abstract

This invention features permeability enhancer molecules, and methods, to increase membrane permeability of various molecules, such as nucleic acids, polynucleotides, oligonucleotides, enzymatic nucleic acid molecules, antisense nucleic acid molecules, 2-5A antisense chimeras, triplex forming oligonucleotides, decoy RNAs, dsRNAs, siRNAs, aptamers, or antisense nucleic acids containing nucleic acid cleaving chemical groups, peptides, polypeptides, proteins, carbohydrates, steroids, metals and small molecules, thereby facilitating cellular uptake of such molecules.

We claim:  
     
         1 . A cationic lipid having the formula II:  
       
         
           
           
               
               
           
         
       
       wherein, n is 1, 2 or 3 carbon atoms; n 1  is 2, 3, 4 or 5 carbon atoms; R and R 1  independently represent C12-C22 alkyl chain which are saturated or unsaturated, wherein the unsaturation is represented by 1-4 double bonds; and Alk represents methyl, hydroxyalkyl or a combination thereof.  
     
     
         2 . A cationic lipid having the formula III:  
       
         
           
           
               
               
           
         
       
       wherein, R and R 1  independently represent C12-C22 alkyl chain which are saturated or unsaturated, wherein the unsaturation is represented by 1-4 double bonds; and R 2  is H, PEG, acyl or alkyl.  
     
     
         3 . A cationic lipid having the formula IV:  
       
         
           
           
               
               
           
         
       
       wherein, n is 1-6 carbon atoms; R and R 1  independently represent C12-C22 alkyl chain which are saturated or unsaturated, wherein the unsaturation is represented by 1-4 double bonds; and R 2  is H, PEG, carboxamidine, alkyl, acyl, aryl, substituted carboxamidine,  
       
         
           
           
               
               
           
         
       
       wherein R 3  is H, or PO 3 H 2  and R 4  is OH, NH 2  or ═O.  
     
     
         4 . A cationic lipid having the formula V:  
       
         
           
           
               
               
           
         
       
       wherein, n is 1-6 carbon atoms; X and X 1  independently represent C12-C22 alkyl chain which are saturated or unsaturated, wherein the unsaturation is represented by 1-4 double bonds; B is a nucleic acid base or H; and R 5  is H, PEG, or carboxamidine.  
     
     
         5 . A cationic lipid having the formula VI:  
       
         
           
           
               
               
           
         
       
       wherein, 
 n is 1, 2 or 3 carbon atoms;  
 R and R 1  independently represent C12-C22 alkyl chain which are saturated or unsaturated, wherein the unsaturation is represented by 1-4 double bonds; and R 2  and R 3  is independently H, polyethylene glycol (PEG) or  
                     
 
     
     
         6 . A cationic lipid having the formula VII: 
 R 6 -L 1 -Cholesterol    wherein,    R 6  is selected from the group consisting of arginyl methyl ester, arginyl amide, homoarginyl methyl ester, homoarginyl amide, ornithine methyl ester, ornithine amide, lysyl methyl ester, lysyl amide, triethylenetetramine (TREN), N,N′-di-carboxamidine TREN, N-benzyl histidyl methyl ester, pyridoxyl and aminopropylimidazole; and    L 1  is a linker represented by R 7 PO2, wherein R 7  is H, CH 3 , or CH 2 CH 3 .    
     
     
         7 . A cationic lipid having the formula VIII: 
 R 8 -L 2 -Cholesterol    wherein, R 8  is selected from the group consisting of arginyl, N-Boc arginyl, homoarginyl, N-Boc homoarginyl, ornithine, N-Boc ornithine, N-benzyl histidyl, lysyl, N-Boc lysyl, N-methyl arginyl, N-methyl guanidine, guanidine and pyridoxyl; and L 2  is a linker represented by NH, glycine, N-butyldiamine or guanidine.    
     
     
         8 . The cationic lipid of  claim 1 , wherein said cationic lipid is N′-palmityl-N′-oleyl-alpha, gamma-bis-trimethylammoniumbutyryl-glycinamide.  
     
     
         9 . The cationic lipid of  claim 2 , wherein said cationic lipid is N′-palmityl-N′-oleyl-N-carboxamidine-glycinamide.  
     
     
         10 . The cationic lipid of  claim 2 , wherein said cationic lipid is N′-palmityl-N′-oleyl-guanidine.  
     
     
         11 . The cationic lipid of  claim 7 , wherein said cationic lipid is Boc arginine-cholesterylamide.  
     
     
         12 . The cationic lipid of  claim 6 , wherein said cationic lipid is Cholesterol-Lysine-methyl ester-methylphosphonoamidate.  
     
     
         13 . The cationic lipid of  claim 6 , wherein said cationic lipid is Cholesterol-methyl ester-homoarginine-methylphosphonoamidate.  
     
     
         14 . The cationic lipid of  claim 6 , wherein said cationic lipid is Cholesterol-Lysine-amide-methylphosphonoamidate.  
     
     
         15 . The cationic lipid of  claim 6 , wherein said cationic lipid is Cholesterol-TREN-methylphosphonoamidate.  
     
     
         16 . The cationic lipid of  claim 6 , wherein said cationic lipid is Cholesterol-TREN-bis-guanidinium methylphosphonoamidate.  
     
     
         17 . The cationic lipid of  claim 6 , wherein said cationic lipid is Cholesterol-histidine-methylphosphonoamidate.  
     
     
         18 . The cationic lipid of  claim 6 , wherein said cationic lipid is Cholesterol-aminopropylimidazole-methylphosphonoamidate.  
     
     
         19 . The cationic lipid of  claim 3 , wherein said cationic lipid is N′-palmityl-N′-oleyl-beta-alanine amide.  
     
     
         20 . The cationic lipid of  claim 3 , wherein said cationic lipid is N′-palmityl-N′-oleyl-N-carboxamidine beta-alanine amide.  
     
     
         21 . The cationic lipid of  claim 3 , wherein said cationic lipid is N(N′-palmityl-N′-oleyl-amidopropyl)pyridoxamine.  
     
     
         22 . The cationic lipid of  claim 7 , wherein said cationic lipid is N-cholesteryl-pyridoxamine.  
     
     
         23 . The cationic lipid of  claim 7 , wherein said cationic lipid is 3′,5′-Di-palmitoyl-2′-deoxy-2′-(N-carboxamidine-beta-alanineamido) uridine.  
     
     
         24 . The cationic lipid of any of claims  1 - 7 , wherein said cationic lipid ion-pairs to a negatively charged polymer selected from the group consisting of RNA, DNA, and protein.  
     
     
         25 . The cationic lipid of  claim 24 , wherein said RNA and said DNA comprise one or more modifications.  
     
     
         26 . The cationic lipid of  claim 24 , wherein said RNA is an enzymatic RNA.  
     
     
         27 . The cationic lipid of  claim 24 , wherein said DNA is an enzymatic DNA.  
     
     
         28 . A cationic lipid having the formula IX:  
       
         
           
           
               
               
           
         
         wherein R is independently C12-C22 alkyl chain which are saturated or unsaturated, wherein the unsaturation is represented by 1-4 double bonds; and R 1  is selected from the group consisting of TREN, N,N′-di-carboxamidine TREN, lysyl, arginyl, omithyl, homoarginyl, histidyl, aminopropylimidazole, and spermine carboxylic acid.  
       
     
     
         29 . The cationic lipid of  claim 28 , wherein said cationic lipid is N 2 ,N 3 -di-oleyl-(N,N′-diguanidinoethyl-aminoethane)-2,3-diaminopropionic acid.  
     
     
         30 . The cationic lipid of  claim 4 , wherein said cationic lipid is 3′,5′-Di-palmitoyl-2′-deoxy-2′(beta-alanineamido) uridine.  
     
     
         31 . The cationic lipid of  claim 4 , wherein said cationic lipid is 3′,5′-Di-palmitoyl-2′-deoxy-2′(N-carboxamidine-beta-alanineamido) uridine.  
     
     
         32 . The cationic lipid of any of claims  1 - 7 , wherein said cationic lipid is linked to polyethylene glycol (PEG).  
     
     
         33 . The cationic lipid of  claim 32 , wherein said PEG is between about 2000-5000 daltons inclusive.