Novel triazine-based detoxification agents and their use

Abstract

An affinity ligand-matrix conjugate comprises the matrix and conjugated thereto by the group Z, a ligand having general formula (I): wherein one X is N and the other X is N, CCL or CCn; A 1 and A 2 are each independently O, S or N—R 1 and R 1 is H, C 1-6 alkyl, C 1-6 hydroxyalkyl, benzyl or β-phenylethyl; B 1 and B 2 are each independently an optionally substituted hydrocarbon linkage containing from 1 to 10 carbon atoms; D 1 is H or a primary amino, secondary amino, tertiary amino, quaternary ammonium, imidazole, guanidine or amidino group; and D 2 is a secondary amino, tertiary amino, quaternary ammonium, imadazole, guanidine or amidino group; or B 2 -D 2 is —CHCOOH—(CH 2 ) 3-4 —NH 2 ; and p is 0 or 1 . Such conjugates are useful for the separation, isolation, purification, characterization, identification or quantification of an endotoxin.

1 . An affinity ligand-matrix conjugate comprising the matrix and, conjugated thereto by the group Z, a ligand having the general formula (I):  
       
         
           
           
               
               
           
         
       
       wherein one X is N and the other X is N, CCl or CCN; 
 A 1  and A 2  are each independently O, S or N—R 1  and R 1  is H, C 1-6  alkyl, C 1-6  hydroxyalkyl, benzyl or β-phenylethyl;  
 B 1  and B 2  are each independently an optionally substituted hydrocarbon linkage containing from 1 to 10 carbon atoms;  
 D 1  is H or a primary amino, secondary amino, tertiary amino, quaternary ammonium, imidazole, guanidine or amidino group; and  
 D 2  is a secondary amino, tertiary amino, quaternary ammonium, imidazole, guanidine or amidino group; or  
 B 2 -D 2  is —CHCOOH—(CH 2 ) 3-4 —NH 2 ; and  
 p is 0 or 1;  
 with the proviso that, when each X is N, A 1  is NR 1 , A 2  is NH, B 2  is phenyl, D 1  is H, D 2  is amidino and p is zero, then R 1  is not methyl, β-phenylethyl or β-hydroxyethyl.  
 
     
     
         2 . The conjugate according to  claim 1 , wherein A 1  and A 2  are each independently N—R 1  wherein R 1  is H, C 1-6  alkyl, C 1-6  hydroxyalkyl, benzyl or β-phenylethyl.  
     
     
         3 . The conjugate according to  claim 2 , wherein A 1  and A 2  are each NH.  
     
     
         4 . The conjugate according to  claim 1 , wherein B 1  and B 2  are each independently —CHCOOH—(CH 2 ) 1-4 — or a divalent ethyl, propyl, 2-hydroxypropyl, butyl, pentyl, hexyl, phenyl, naphthyl or cyclohexyl group.  
     
     
         5 . The conjugate according to  claim 4 , wherein B 1  and B 2  are each independently —CHCOOH—(CH 2 ) 3-4 — or a divalent butyl, pentyl or phenyl group.  
     
     
         6 . The conjugate according to  claim 4 , wherein B 1  and B 2  are each independently —CHCOOH—(CH 2 ) 3-4 — and p is 1.  
     
     
         7 . The conjugate according to  claim 1 , wherein D 1  is H, amino, imidazolyl, guanidine, aminidino, trimethylammonium, triethylammonium, dimethylamino, diethylamino, methylamino or ethylamino.  
     
     
         8 . The conjugate according to  claim 1 , wherein D 2  is imidazolyl, guanidine, aminidino, trimethylammonium, triethylammonium, dimethylamino, diethylamino, methylamino or ethylamino.  
     
     
         9 . The conjugate according to  claim 1 , wherein p is 1.  
     
     
         10 . The conjugate according to  claim 1 , wherein each X is N.  
     
     
         11 . The conjugate according to  claim 1 , wherein Z is  
         -T-[L-V] m -   (II)  
       wherein T is O, S or NR 2  and R 2  is H or C 1-6  alkyl; 
 V is O, S, —COO—, CONH, NHCO, —PO 3 H—, NH-arylene-SO 2 —CH 2 —CH 2 — or N—R 3  and R 3  is H or C 1-6  alkyl;  
 L is an optionally substituted hydrocarbon linkage containing from 2 to 20 carbon atoms; and  
 m is 0 or 1.  
 
     
     
         12 . The conjugate according to  claim 11 , wherein R 2  and R 3  are each H.  
     
     
         13 . The conjugate according to  claim 11 , wherein T is O or NH.  
     
     
         14 . The conjugate according to  claim 13 , wherein T is NH.  
     
     
         15 . The conjugate according to  claim 1 , wherein m is 1 and L is a divalent butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl or dodecyl group.  
     
     
         16 . The conjugate according to  claim 11 , wherein m is 1 and V is O, —CONH—, —NHCO— or N—R 3 .  
     
     
         17 . The conjugate according to  claim 16 , wherein V is O or NH.  
     
     
         18 . The conjugate according to  claim 17 , wherein V is NH.  
     
     
         19 . The conjugate according to  claim 17 , wherein Z-M (M being the matrix) is —NH—(CH 2 ) 4-10 —NH—M.  
     
     
         20 . The conjugate according to  claim 19 , selected from any of formulae XXX to XXXX.  
     
     
         21 . The conjugate according to  claim 1 , wherein the matrix is optionally tresyl-activated, sulphonyl chloride-activated, tosyl-activated, vinyl sulphone-activated or epoxy-activated.  
     
     
         22 . An affinity ligand of general formula (I):  
       
         
           
           
               
               
           
         
       
       wherein one X is N and the other X is N, CCl or CCN; 
 A 1  and A 2  are each independently O, S or N—R, and R 1  is H, C 1-6  alkyl, C 1-6  hydroxyalkyl, benzyl or β-phenylethyl;  
 B 1  and B 2  are each independently an optionally substituted hydrocarbon linkage containing from 1 to 10 carbon atoms;  
 D 1  is H or a primary amino, secondary amino, tertiary amino, quaternary ammonium, imidazole, guanidine or amidino group; and  
 D 2  is a secondary amino, tertiary amino, quaternary ammonium, imidazole, guanidine or amidino group; or  
 B 2 -D 2  is —CHCOOH—(CH 2 ) 3-4 —NH 2 ; and  
 p is 0 or 1; and  
 wherein Z is a functional group capable of reaction with a solid matrix; and with the proviso that, when each X is N, A 1  is NR 1 , A 2  is NH, B 2  is phenyl, D 1  is H, D 2  is amidino and p is zero, then R 1  is not methyl, β-phenylethyl or β-hydroxyethyl.  
 
     
     
         23 . The ligand according to  claim 22 , wherein the matrix is optionally tresyl-activated, sulphonyl chloride-activated, tosyl-activated, vinyl sulphone-activated or epoxy-activated.  
     
     
         24 . The ligand according to  claim 22 , wherein Z is F, Cl, Br or I.  
     
     
         25 . The ligand according to  claim 22 , wherein Z is —NH—(CH 2 ) 2-20 —NH 2 .  
     
     
         26 . The ligand according to  claim 22 , wherein Z is —NH—(CH 2 ) 2-20 —(CO) 0-1 —OH.  
     
     
         27 . The ligand according to  claim 22 , wherein Z is  
         -T-[L-V] m -R 10 —CR 11 ═CH 2    
       wherein T is O, S or NR 2  and R 2  is H or C 1-6  alkyl; 
 V is O, S, —COO—, CONH, NHCO, —PO 3 H—, NH-arylene-SO 2 —CH 2 —CH 2 — or N—R 3  and R 3  is H or C 1-6  alkyl;  
 L is an optionally substituted hydrocarbon linkage containing from 2 to 20 carbon atoms;  
 m is 0 or 1;  
 R 10  is CO, CH 2 , NH—CH 2 — or —S—CH 2 —; and  
 R 11  is H or C 1-6  alkyl.  
 
     
     
         28 . The ligand according to  claim 24 , wherein Z is Cl, and each X is N.  
     
     
         29 . The ligand according to  claim 28 , wherein each X is N.  
     
     
         30 . The ligand according to  claim 22 , selected from any of formulae XVI to XXVI.  
     
     
         31 . A method for the separation, isolation, purification, characterization, identification or quantification of an endotoxin; wherein said method utilizes an affinity ligand or conjugate having the general formula (I):  
       
         
           
           
               
               
           
         
       
       wherein one X is N and the other X is N, CCl or CCN; 
 Z is a functional group capable of reaction with a solid matrix;  
 A 1  and A 2  are each independently O, S or N—R 1  and R 1  is H, C 1-6  alkyl, C 1-6  hydroxyalkyl, benzyl or β-phenylethyl;  
 B 1  and B 2  are each independently an optionally substituted hydrocarbon linkage containing from 1 to 10 carbon atoms;  
 D 1  is H or a primary amino, secondary amino, tertiary amino, quaternary ammonium, imidazole, guanidine or amidino group; and  
 D 2  is a secondary amino, tertiary amino, quaternary ammonium, imidazole, guanidine or amidino group; or  
 B 2 -D 2  is —CHCOOH—(CH 2 ) 3-4 —NH 2 ; and  
 p is 0 or 1.  
 
     
     
         32 . The method according to  claim 31 , for the removal of an endotoxin from water or an aqueous solution, body fluid, blood, plasma, solution of pharmaceutical products, protein or other compound of biological origin.  
     
     
         33 . The method according to  claim 32 , which comprises using the conjugate for the extracorporeal removal of endotoxin from whole blood or plasma, prior to re-infusion into the door or another recipient.  
     
     
         34 . The method according to  claim 31 , wherein the endotoxin originates from a Gram-negative bacterium.  
     
     
         35 . The method according to  claim 31 , in an endotoxin-containing solution or liquid is applied to the conjugate at a pH of 1.0 to 13.0.