摘要
目的采用膜电位高通量筛选技术进行化合物库筛选及活性研究,以期发现不同于苯二氮[艹卓]类结构的新型催眠物质。方法采用高通量实时荧光膜电位检测方法对化合物库进行筛选,获得候选化合物;通过膜片钳电生理和放射配基受体结合检测技术进行体外活性评价;采用戊巴比妥钠阈下睡眠检测和小鼠自发活动进行体内活性评价。结果通过膜电位检测方法从2288个化合物中筛选得到EC50值低于阳性对照地西泮[EC50=(4.43±0.41)μmol·L^-1]的化合物A11[EC50=(1.98±0.47)μmol·L^-1];通过膜片钳检测其体外活性EC50=(20.47±0.45)μmol·L^-1;[3H]-氟硝西泮受体结合检测技术表明该化合物为非苯二氮[艹卓]类化合物;戊巴比妥钠阈下睡眠时间检测实验表明化合物A11可明显缩短睡眠潜伏期(P<0.05),延长睡眠持续时间;A11在0.5、1、2 mg·kg^-1剂量下对90 min后小鼠自发活动有明显抑制作用(P<0.05,P<0.01)。结论本研究基于GABAAR高通量筛选获得的化合物A11具有较强的睡眠活性,可能是一种不同于苯二氮[艹卓]类药物作用位点的新型催眠物质。
Objective To screen the activity of compounds by membrane potential high-throughput screening technology,and to discover new structure sleeping drugs with different from benzodiazepines.Methods High-throughput real-time fluorescent membrane potential detection method was used to screen the compound library to obtain candidate compounds.Patch clamp electrophysiology and radioligand receptor binding detection technology were used to evaluate the activity of compound in vitro.Pentobarbital sodium subthreshold sleep detection and spontaneous activities of mice were used to evaluate the activity of compounds in vivo.Results The EC50[(1.98±0.47) μmol·L^-1]of A11 screened from 2288 compounds by membrane potential detection method was lower than that of the positive control diazepam[(4.43±0.41) μmol·L^-1].The EC50 was (20.47±0.45) μmol·L^-1 by patch clamp electrophysiology.[3H]-Flunitrazepam receptor binding assay showed that A11 was a non-benzodiazepine-site compound.The subthreshold sleep time test of pentobarbital sodium showed that A11 could significantly shorten the sleep latency (P < 0.05) and prolong the sleep duration.The spontaneous activities of mice were significantly inhibited by A11 at the doses of 0.5,1 and 2 mg·kg-1 (P<0.05,P < 0.01).Conclusion A11 obtained by high-throughput screening based on GABAAR shows strong sleep activity,which may be a new sleeping substance different from the action site of benzodiazepines.
作者
张毅
石童
陈学军
张瑞华
邓诗坤
崔雅岚
时梦
徐建富
王陈
李丽琴
ZHANG Yi;SHI Tong;CHEN Xue-jun;ZHANG Rui-hua;DENG Shi-kun;CUI Ya-lan;SHI Meng;XU Jian-fu;WANG Chen;LI Li-qin(Research Institute of Chemical Defense,Academy of Military Sciences,Beijing 102205)
出处
《中南药学》
CAS
2020年第12期1945-1949,共5页
Central South Pharmacy
