Novel triazine-based detoxification agents and their use
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.