首页|联用多步骤虚拟筛选方法发现具有新母核的GABAA受体正性变构调节剂

联用多步骤虚拟筛选方法发现具有新母核的GABAA受体正性变构调节剂

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GABAA受体主要介导哺乳动物中枢神经系统的抑制性信号传递,是镇静催眠药的关键靶点.在寻找具有新母核的镇静催眠药的过程中,计算机辅助药物设计(CADD)方法显示出巨大的优势.在这项研究中,首先,我们通过机器学习模型、分子对接模型和分子力学广义玻恩比表面积(MMGBSA)方法筛选了来自于商业数据库中的41112种化合物.经过筛选,我们得到了16个化合物,然后我们通过全细胞膜片钳电生理学实验验证了4个结构新颖的化合物确实为有效的GABAA受体正性变构调节剂.其中,化合物GPR120在细胞水平和动物水平都得到了实验验证.在重组表达α1β2γ2型受体的皮层神经元中,在10和50 µmol∙L-1浓度下,GPR120可将GABA EC3-10电流分别增强71.5%和163.8%.通过全分解贡献分析和点突变实验,我们发现GPR120与GABAA受体结合的关键位点是H102,与阳性药物地西泮相似.为了进一步验证GPR120在动物水平上的功能,我们进行了运动活动测试和翻正反射消失(LORR)实验.GPR120对小鼠的运动活动有抑制作用,6 h后可恢复,说明GPR120是一种中度镇静剂.在戊巴比妥钠(PB)诱导的翻正反射消失实验中,与生理盐水组相比,GPR120(20 mg∙kg-1)可显著缩短开始LORR的时间并延长LORR的持续时间.综上所述,通过联用多种虚拟筛选方法,我们发现了GPR120是一种具有新型母核的中度强度镇静剂.
Identification of Novel GABAA Receptor Positive Allosteric Modulators with Novel Scaffolds via Multistep Virtual Screening
The GABAA receptor mainly mediates inhibitory signal transmission in mammalian central nervous systems and is the key target of sedative-hypnotics.However,the long-term use of sedative-hypnotics often leads to drug resistance,necessitating the development of novel sedative-hypnotics.This development can be achieved with novel scaffolds designed via the computer-aided drug design methods to obtain significant advantages.In this study,robust virtual screening models were established by identifying effective positive allosteric modulators of the GABAA receptor from ChEMBL and BindingDB databases.These compounds combined with randomly extracted negative compounds were firstly applied for a 10-fold cross validation and grid search to establish machine learning models which were subsequently evaluated in an independent test set.In this step,4 machine learning methods and 6 fingerprints were used to establish 24 models.In the test set,the CDK_LR model performed the best(MCC = 0.751)and was used for subsequent virtual screening.Two effective molecular docking models were also established based on conformation 6D6T and 6D6U,wherein the root mean square deviation(RMSD)values of redocking experiments were 1.141 and 1.505 Å(1 Å = 0.1 nm),respectively.During the virtual screening,41112 compounds from a commercial database were scanned by machine learning,molecular docking,and molecular mechanics-generalized Born surface area models.After the screening,16 hits were obtained,4 of which were structurally novel positive hits verified by whole-cell patch-clamp electrophysiology experiments.The compound GPR120 was verified experimentally at both the cell and animal levels.In cortical neurons recombinantly expressing α1β2γ2-type receptors,at 10 and 50 µmol∙L-1,GPR120 could potentiate GABA EC3-10 current by 71.5%and 163.8%,respectively.Total decomposition contribution analysis and point mutation experiment showed that the key binding site between GPR120 and the GABAA receptor is H102,similar to that of the positive drug Diazepam.To further verify GPR120 function at the animal level,locomotor activity and loss of righting reflex(LORR)tests were performed.GPR120 inhibited the locomotor activity of mice,which recovered after 6 h,indicating that GPR120 is a moderate sedative.In the pentobarbital sodium-induced righting reflex hour test,GPR120(20 mg∙kg-1)significantly shortened the time to start LORR and prolonged its duration compared with the saline control group.In summary,using integrated virtual screening methods,GPR120 was identified as a moderate sedative with a novel scaffold.

Machine learningIon channelMolecular dockingVirtual screeningGABAA receptor

孔维恺忻、廉靖靖、彭超、朱杰、郑钰琳、黄巍然、张博文、段桂芳、马琳、彭晓东、马维宁、朱素杰、黄卓

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北京大学医学部药学院分子与细胞药理学系,北京 100191

宁夏医科大学基础医学院,宁夏 750004

赫尔辛基大学分子医学研究所,赫尔辛基 00250,芬兰

三驱科技(杭州)有限公司,杭州 310012

青岛大学附属医院转化医学研究所,山东 青岛 266023

康迈迪森(北京)医药科技有限公司,北京 100094

中国医科大学附属盛京医院神经外科,沈阳 110022

北京大学医学部天然药物与仿生药物国家重点实验室,北京 100191

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机器学习 离子通道 分子对接 虚拟筛选 GABAA受体

国家自然科学基金国家自然科学基金国家自然科学基金北京大学肿瘤医院科学基金中国脑科学与类脑智能技术国家计划山东省高等学校青创团队计划宁夏回族自治区重点研发计划

819035393200067482271498JC2023042021ZD02021022022KJ1452022BEG02042

2024

10.3866/PKU.WHXB202302044

物理化学学报
中国化学会

物理化学学报

CSTPCD北大核心
影响因子：0.951
ISSN：1000-6818
年,卷(期)：2024.40(1)