Key Laboratory of Public Health Detection and Etiological Research of Zhejiang Province, Department of Microbiology, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, Zhejiang, 310051, People's Republic of China.
College of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 310053, People's Republic of China.
Drug Des Devel Ther. 2024 Mar 2;18:651-665. doi: 10.2147/DDDT.S444625. eCollection 2024.
This study aims to investigate the in vitro antiviral effects of the aqueous solution of Changyanning (CYN) tablets on Enterovirus 71 (EV71), and to analyze its active components.
The in vitro anti-EV71 effects of CYN solution and its herbal ingredients were assessed by testing the relative viral RNA (vRNA) expression level and the cell viability rates. Material basis analysis was performed using HPLC-Q-TOF-MS/MS detection. Potential targets and active components were identified by network pharmacology and molecular docking. The screened components were verified by in vitro antiviral experiments.
CYN solution exerted anti-EV71 activities as the vRNA is markedly reduced after treatment, with a half maximal inhibitory concentration (IC) of 996.85 μg/mL. Of its five herbal ingredients, aqueous extract of (AEMC) and leaves of Hance (AELLF) significantly inhibited the intracellular replication of EV71, and the IC was tested as 202.57 μg/mL and 174.77 μg/mL, respectively. Based on HPLC-Q-TOF-MS/MS results, as well as the comparison with the material basis of CYN solution, a total of 44 components were identified from AEMC and AELLF. Through network pharmacology, AKT1, ALB, and SRC were identified as core targets. Molecular docking performed between core targets and the components indicated that 21 components may have anti-EV71 effects. Of these, nine were selected for in vitro pharmacodynamic verification, and only rosmarinic acid manifested in vitro anti-EV71 activity, with an IC of 11.90 μg/mL. Moreover, rosmarinic acid can stably bind with three core targets by forming hydrogen bonds.
CYN solution has inhibitory effects on EV71 replication in vitro, and its active component was identified as rosmarinic acid. Our study provides a new approach for screening and confirmation of the effective components in Chinese herbal preparation.
本研究旨在探讨肠炎宁片水溶液对肠道病毒 71 型（EV71）的体外抗病毒作用，并分析其活性成分。
采用检测相对病毒 RNA（vRNA）表达水平和细胞活力率的方法，评估肠炎宁溶液及其草药成分的抗 EV71 作用。采用 HPLC-Q-TOF-MS/MS 检测进行物质基础分析。通过网络药理学和分子对接鉴定潜在靶点和活性成分。通过体外抗病毒实验验证筛选出的成分。
肠炎宁溶液对 EV71 具有抗病毒活性，因为处理后 vRNA 明显减少，半数最大抑制浓度（IC）为 996.85μg/mL。其五种草药成分中，（AEMC）和（AELLF）的水提物显著抑制 EV71 的细胞内复制，IC 分别为 202.57μg/mL 和 174.77μg/mL。基于 HPLC-Q-TOF-MS/MS 结果以及与肠炎宁溶液物质基础的比较，从 AEMC 和 AELLF 中总共鉴定出 44 种成分。通过网络药理学，鉴定出 AKT1、ALB 和 SRC 为核心靶点。对核心靶点与成分之间的分子对接表明，有 21 种成分可能具有抗 EV71 作用。其中，有 9 种被选择进行体外药效验证，只有迷迭香酸表现出体外抗 EV71 活性，IC 为 11.90μg/mL。此外，迷迭香酸可以通过形成氢键与三个核心靶点稳定结合。
肠炎宁溶液对 EV71 在体外具有抑制作用，其活性成分被鉴定为迷迭香酸。本研究为筛选和确认中药制剂有效成分提供了一种新方法。
