SONG ET AL.
4
7 of 8
|
3
(d, Jc‐f = 3.2 Hz), 131.2 (d, Jc‐f = 8.2 Hz), 130.7, 118.6, 118.5, 115.6
4‐(2‐(4‐Methylbenzylthio)benzo[d]oxazol‐6‐yl)‐2H‐1,2,4‐triazol‐
3(4H)‐one (5l)
2
(d, Jc‐f = 21.4 Hz), 104.4, 35.5. IR (KBr) cm−1: 3144 (N‐H), 1711
(C=O). ESI‐HRMS calcd for C16H12FN4O2S+ ([M+H]+): 343.0660;
Mp 224°C, yield 75%. H‐NMR (CDCl3+DMSO‐d6, 300 MHz): δ 2.25
(s, 3H, Ar‐CH3), 4.46 (s, 2H, SCH2), 7.07 (d, 2H, J = 7.9 Hz, benzyl‐H),
7.28 (d, 2H, J = 7.9 Hz, benzyl‐H), 7.44 (dd, 1H, J1 = 8.5 Hz, J2 = 2.0 Hz,
Ph‐H), 7.57 (d, 1H, J = 8.5 Hz, Ph‐H), 7.81 (d, 1H, J = 2.0 Hz, Ph‐H),
7.93 (s, 1H, CH=N), 11.82 (s, 1H, CO‐NH). 13H‐NMR (CDCl3+DMSO‐
d6, 75 MHz): δ 165.7, 153.6, 151.7, 140.5, 137.0, 135.8, 132.8, 130.3,
129.4, 129.0, 118.6, 118.5, 104.6, 36.3, 21.2. IR (KBr) cm−1: 3199
(N‐H), 1713 (C=O). ESI‐HRMS calcd for C17H15N4O2S+ ([M+H]+):
339.0910; found: 339.0906.
found: 343.0660.
4‐(2‐(2‐Chlorobenzylthio)benzo[d]oxazol‐6‐yl)‐2H‐1,2, 4‐triazol‐
3(4H)‐one (5h)
Mp 201–203°C, yield 63%. H‐NMR (CDCl3+DMSO‐d6, 300 MHz): δ
4.59 (s, 2H, SCH2), 7.18–7.23 (m, 2H, benzyl‐H), 7.33 (d, 1H,
J = 7.5 Hz, benzyl‐H), 7.44 (d, 1H, J = 7.6 Hz, benzyl‐H), 7.57 (dd, 1H,
J1 = 8.6 Hz, J2 = 2.2 Hz, Ph‐H), 7.60 (d, 1H, J = 8.6 Hz, Ph‐H), 7.81 (d,
1H, J = 2.2 Hz, Ph‐H), 7.93 (s, 1H, CH=N), 11.89 (s, 1H, CO‐NH). 13H‐
NMR (CDCl3+DMSO‐d6, 75 MHz): δ 165.5, 153.6, 151.8, 140.6,
135.9, 134.1, 133.8, 133.2, 131.4, 130.3, 129.7, 127.3, 118.7, 118.7,
104.7, 34.3. IR (KBr) cm−1: 3205 (N‐H), 1711 (C=O). ESI‐HRMS calcd
for C16H12ClN4O2S+ ([M+H]+): 359.0364; found: 359.0369.
4‐(2‐(4‐Methoxybenzylthio)benzo[d]oxazol‐6‐yl)‐2H‐1,2,4‐triazol‐
3(4H)‐one (5m)
Mp 202–206°C, yield 73%. H‐NMR (CDCl3+DMSO‐d6, 300 MHz): δ
1.17 (s, 3H, Ar‐OCH3), 4.47 (s, 2H, SCH2), 7.23 (d, 2H, J = 8.2 Hz,
benzyl‐H), 7.38 (d, 2H, J = 8.2 Hz, benzyl‐H), 7.45 (d, 1H, J = 8.5 Hz,
Ph‐H), 7.57 (d, 1H, J = 8.5 Hz, Ph‐H), 7.82 (s, 1H, Ph‐H), 7.94 (s, 1H,
CH=N), 11.83 (s, 1H, CO‐NH). 13H‐NMR (CDCl3+DMSO‐d6, 75 MHz):
δ 165.3, 153.5, 151.8, 140.4, 135.9, 135.2, 133.4, 130.7, 130.5, 128.8,
118.7, 118.5, 104.5, 35.5, 29.6. IR (KBr) cm−1: 3200 (N‐H), 1713
(C=O). ESI‐HRMS calcd for C17H15N4O3S+ ([M+H]+): 355.0859;
found: 355.0859.
4‐(2‐(3‐Chlorobenzylthio)benzo[d]oxazol‐6‐yl)‐2H‐1,2,4‐triazol‐
3(4H)‐one (5i)
Mp 196–197°C, yield 72%. H‐NMR (CDCl3+DMSO‐d6, 300 MHz): δ
4.45 (s, 2H, SCH2), 7.18–7.31 (m, 3H, benzyl‐H), 7.41 (s, 1H, benzyl‐
H), 7.40 (dd, 1H, J1 = 8.5 Hz, J2 = 1.9 Hz, Ph‐H), 7.57 (d, 1H, J = 8.5 Hz,
Ph‐H), 7.78 (d, 1H, J = 1.9 Hz, Ph‐H), 7.82 (s, 1H, CH=N), 11.76 (s, 1H,
CO‐NH). 13H‐NMR (CDCl3+DMSO‐d6, 75 MHz): δ 165.3, 153.6,
151.8, 140.6, 138.2, 135.6, 134.2, 130.3, 130.1, 129.0, 128.0, 127.4,
118.7, 118.6, 104.7, 35.7. IR (KBr) cm−1: 3206 (N‐H), 1710 (C=O).
ESI‐HRMS calcd for C16H12ClN4O2S+ ([M+H]+): 359.0364; found:
359.0361.
|
4.2
Pharmacology
|
4.2.1
Animals and experimental conditions
Experiments were carried out on Kunming mice (half male and
female) weighing 18–24 g. The mice were housed collectively in
polycarbonate cages in groups of 10, where they were maintained on
a 12‐hr light/dark cycle in a temperature‐controlled (25 2°C)
laboratory with free access to food and water. Each animal was
used only once. Procedures involving animals and their care were
conducted in accordance with the Guide for the Care and Use of
Laboratory Animals, 8th Edition, National Academies Press, Washing-
ton, DC. Local ethical committee approval was also obtained (No.
20170707). In addition, all efforts were made to minimize animal
suffering and to use only the number of animals necessary to produce
reliable scientific data. Electroconvulsions were produced by an
electric stimulation generator (JTC‐1, Chengdu, China). The following
drugs were used in this study: pentylenetetrazole, 3‐MP, TSC, and
BIC (these drugs were obtained from Aladdin Industrial Inc.,
Shanghai, China), valproate sodium, carbamazepine, and phenytoin
(these drugs were obtained from Melongpharma, Dalian, China).
4‐(2‐(4‐Chlorobenzylthio)benzo[d]oxazol‐6‐yl)‐2H‐1,2,4‐triazol‐
3(4H)‐one (5j)
Mp 235–237°C, yield 63%. H‐NMR (CDCl3+DMSO‐d6, 300 MHz): δ
4.46 (s, 2H, SCH2), 6.79 (d, 2H, J = 8.4 Hz, benzyl‐H), 7.32 (d, 2H,
J = 8.4 Hz, benzyl‐H), 7.47 (dd, 1H, J1 = 8.5 Hz, J2 = 1.9 Hz, Ph‐H), 7.58
(d, 1H, J = 8.5 Hz, Ph‐H), 7.77 (d, 1H, J = 1.9 Hz, Ph‐H), 7.99 (s, 1H,
CH=N), 11.85 (s, 1H, CO‐NH). 13H‐NMR (CDCl3+DMSO‐d6, 75 MHz):
δ 165.7, 159.3, 153.6, 151.7, 140.6, 135.9, 130.4, 130.3, 127.8, 118.6,
118.5, 114.2, 104.5, 36.1. IR (KBr) cm−1: 3208 (N‐H), 1711 (C=O).
ESI‐HRMS calcd for C16H12ClN4O2S+ ([M+H]+): 359.0364; found:
359.0360.
4‐(2‐(2,4‐Dichlorobenzylthio)benzo[d]oxazol‐6‐yl)‐2H‐1,2,4‐triazol‐
3(4H)‐one (5k)
Mp 190–194°C, yield 70%. H‐NMR (CDCl3+DMSO‐d6, 300 MHz): δ
4.55 (s, 2H, SCH2), 7.17 (dd, 1H, J1 = 8.3 Hz, J2 = 1.5 Hz, benzyl‐H),
7.36 (d, 1H, J = 1.5 Hz, benzyl‐H), 7.45 (dd, 1H, J1 = 8.5 Hz, J2 = 1.7 Hz,
Ph‐H), 7.57 (d, 1H, J = 8.3 Hz, benzyl‐H), 7.58 (d, 1H, J = 8.5 Hz, Ph‐H),
7.77 (d, 1H, J = 1.7 Hz, Ph‐H), 7.93 (s, 1H, CH=N), 11.83 (s, 1H, CO‐
NH). 13H‐NMR (CDCl3+DMSO‐d6, 75 MHz): δ 165.1, 153.6, 151.9,
140.5, 135.7, 134.9, 134.4, 132.8, 132.4, 130.4, 129.4, 127.4, 118.7,
118.5, 104.6, 33.7. IR (KBr) cm−1: 3212 (N‐H), 1714 (C=O). ESI‐HRMS
calcd for C16H11Cl2N4O2S+ ([M+H]+): 392.9974; found: 392.9982.
|
4.2.2
Experimental method
The animal experiments, including the MES test, subcutaneous
pentylenetetrazole‐induced seizure test (sc‐PTZ), the rotarod test,
3‐MP‐induced seizure test, BIC‐induced seizures test, determination
of brain GABA concentrations by ELISA, and test for the effects of
TSC on the anti‐MES action of 5f were carried out according to the
previous report.[13]