麸炒僵蚕汤剂的多肽成分表征及抗GAT-1活性肽的虚拟筛选  

Characterization of peptide components of bran-fried Bombyx batryticatus decoction and virtual screening of anti-GAT-1 active peptides

作　　者：黄雅阳 刘亚雄[2] 萧晓彤 罗卓雅[2] 杨志业 HUANG Ya-yang;LIU Ya-xiong;XIAO Xiao-tong;LUO Zhuo-ya;YANG Zhi-ye(School of Pharmaceutical Sciences,Guangzhou Medical University,Guangzhou 511436,China;Guangdong Institute for Drug Control,NMPA Key Laboratory of Rapid Drug Inspection Technology,Guangdong Biomedical Technology Collaborative Innovation Center,Guangzhou 510663,China)

机构地区：[1]广州医科大学药学院,广州511436 [2]广东省药品检验所,国家药品监督管理局药品快速检验技术重点实验室,广东省生物医药科技协同创新中心,广州510663

出　　处：《药物分析杂志》2024年第8期1356-1364,共9页Chinese Journal of Pharmaceutical Analysis

基　　金：广东省科技厅科技计划项目(2021B1111610005);广东省自然基金面上项目(2022A1515011850);广东省中医药局科研项目(20221056)。

摘　　要：目的:分析麸炒僵蚕汤剂的多肽组成,并虚拟筛选具有GAT-1结合活性的抗癫痫多肽。方法:利用基于微升流速液相色谱-质谱联用(μLC-MS/MS)的多肽组学技术,对麸炒僵蚕汤剂的多肽成分进行广泛鉴定,通过生物信息学和计算机模拟虚拟筛选潜在的抗癫痫活性肽,包括生物活性预测、血脑屏障通透性预测、毒性预测和分子对接,最后,利用分子动力学模拟分析多肽和靶蛋白的相互作用。结果:从麸炒僵蚕汤剂中共鉴定得到384条多肽,从中虚拟筛选出19条结合亲和力高于对照药物的潜在抗癫痫活性多肽,其中,多肽FDHFDFDAF的结合亲和力最高,其次是EHYAWGIK,2条多肽主要通过氢键、疏水和静电相互作用与GAT-1结合,分子动力学模拟研究验证了FDHFDFDAF和EHYAWGIK与GAT-1的结合稳定性。结论:基于多肽组学和计算机虚拟筛选技术有利于快速发现动物药的活性多肽成分,本研究对麸炒僵蚕汤剂抗癫痫活性多肽的研究有着重要参考价值,同时为动物药多肽成分研究提供新思路。Objective:To analyze the peptide components of bran-fried Bombyx batryticatus decoction and virtually screen anti-epileptic peptides with GAT-1 binding activity.Methods:The peptide compounds in bran-fried Bombyx batryticatus decoction were extensively identified by micro-flow liquid chromatography-tandem mass spectrometry(μLC-MS/MS)technology.Potential anti-epileptic peptides were screened through bioinformatics and in silico simulations,including bioactivity prediction,blood-brain barrier permeability prediction,toxicity prediction,and molecular docking.Finally,molecular dynamics simulations were employed to analyze the interactions between peptides and the target protein.Results:A total of 384 peptides were identified from bran-fried Bombyx batryticatus decoction,and 19 peptides with higher binding affinities to GAT-1 than that of the control drug were screened as potential anti-epileptic peptides.The FDHFDFDAF exhibited the highest binding affinity,followed by the EHYAWGIK.Two peptides primarily interacted with GAT-1 through hydrogen bonds,hydrophobic,and electrostatic interactions.Molecular dynamics simulations confirmed the binding stability of FDHFDFDAF and EHYAWGIK to GAT-1.Conclusion:Peptidomics and in silico virtual screening techniques facilitate the rapid discovery of active peptide components in animal-derived traditional Chinese medicine.This study has significant reference value for researching anti-epileptic peptides in bran-fried Bombyx batryticatus decoction,and provides new insights for the study of peptide components in animal-derived traditional Chinese medicine.

关 键 词：微升流速液相色谱-质谱联用 麸炒僵蚕 GAT-1 癫痫 活性肽 多肽组学 虚拟筛选 

分 类 号：R917[医药卫生—药物分析学]