A Biolayer Interferometry-Based SARS-COV-2 Mpro-Targeted Active Ingredients Recognition System: Construction and Application in Ligand Screening From Herbal Medicines.

IF 3 3区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Phytochemical Analysis Pub Date : 2024-11-20 DOI:10.1002/pca.3462

Dai Zhang, Bing Han, Xiao-Fei Chen, Shuai Zhao, Wei-Xia Li, Hui Zhang, Ming-Liang Zhang, Meng-Qi Huo, Yong-Sheng Qiu, Ying-Jie Ren, Yao-Dong Zhang, Xian-Qing Ren, Wei Wang, Jin-Fa Tang

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引用次数: 0

Abstract

Introduction: Drug discovery research targeting SARS-CoV-2 and other emerging pathogens remains critically important. Active compounds derived from plants frequently serve as lead compounds for further drug discovery; however, numerous unrelated chemical constituents in crude extracts may obscure the effective ingredients in LC-MS analysis.

Objective: The aim of this study is to construct a biolayer interferometry (BLI)-based system for recognizing active ingredients that inhibit the main protease (Mpro) of SARS-CoV-2 and to identify the active chemical components binding to Mpro from herbal medicines.

Methodology: We developed a novel FRET fluorogenic probe by linking the amino acid sequences of the fluorescent proteins Lssmorange and mKate2 (Ls-mK). The interaction between traditional Chinese medicine and Mpro was analyzed using BLI. Ultrahigh performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) was employed to analyze the composition of herbal medicines.

Results: Fluorescence detection and spectroscopy confirmed the successful construction of an Mpro inhibitor screening system. Lanqin Oral Liquid (LQL) and Gardeniae fructus exhibited strong inhibitory effects on Mpro. Ten compounds were identified from G. fructus extracts; among them, deacetyl asperulosidic acid methyl ester (DAAME) and Gardoside were found to strongly bind to Mpro, with dissociation constants (KD) of 3.41 μM and 801 nM, respectively. The half-maximal inhibitory concentrations (IC50) of DAAME and Gardoside for Mpro were 27.46 and 13.7 μM, respectively.

Conclusion: This study established a functional Mpro inhibitor screening system. Among the 10 components identified from G. fructus that bind to Mpro, DAAME and Gardoside displayed strong binding and inhibitory activity, indicating their potential as lead compounds for inhibiting SARS-CoV-2 viral replication.

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基于生物层干涉仪的 SARS-COV-2 Mpro 靶向活性成分识别系统：中药配体筛选中的构建与应用

导言：针对 SARS-CoV-2 和其他新出现病原体的药物发现研究仍然至关重要。从植物中提取的活性化合物经常成为进一步发现药物的先导化合物；然而，在 LC-MS 分析中，粗提取物中众多不相关的化学成分可能会掩盖有效成分：本研究旨在构建一个基于生物层干涉仪（BLI）的系统，用于识别抑制 SARS-CoV-2 主要蛋白酶（Mpro）的活性成分，并从中草药中识别与 Mpro 结合的活性化学成分：方法：通过连接荧光蛋白Lssmorange和mKate2（Ls-mK）的氨基酸序列，我们开发了一种新型的FRET荧光探针。利用BLI分析了中药与Mpro之间的相互作用。采用超高效液相色谱-四极杆飞行时间质谱（UPLC-QTOF-MS）分析中药的成分：结果：荧光检测和光谱分析证实成功构建了Mpro抑制剂筛选系统。兰沁口服液（LQL）和栀子果实对Mpro有很强的抑制作用。从栀子提取物中鉴定出10个化合物，其中脱乙酰基金丝桃苷酸甲酯（DAAME）和栀子苷能与Mpro强结合，其解离常数（KD）分别为3.41 μM和801 nM。DAAME 和 Gardoside 对 Mpro 的半最大抑制浓度（IC50）分别为 27.46 μM 和 13.7 μM：本研究建立了一个功能性 Mpro 抑制剂筛选系统。结论：该研究建立了一个功能性 Mpro 抑制剂筛选系统，在从枸杞子中鉴定出的 10 种与 Mpro 结合的成分中，DAAME 和 Gardoside 表现出了很强的结合和抑制活性，表明它们有可能成为抑制 SARS-CoV-2 病毒复制的先导化合物。

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来源期刊

Phytochemical Analysis 生物-分析化学

CiteScore

6.00

自引率

6.10%

发文量

审稿时长

1.7 months

期刊介绍： Phytochemical Analysis is devoted to the publication of original articles concerning the development, improvement, validation and/or extension of application of analytical methodology in the plant sciences. The spectrum of coverage is broad, encompassing methods and techniques relevant to the detection (including bio-screening), extraction, separation, purification, identification and quantification of compounds in plant biochemistry, plant cellular and molecular biology, plant biotechnology, the food sciences, agriculture and horticulture. The Journal publishes papers describing significant novelty in the analysis of whole plants (including algae), plant cells, tissues and organs, plant-derived extracts and plant products (including those which have been partially or completely refined for use in the food, agrochemical, pharmaceutical and related industries). All forms of physical, chemical, biochemical, spectroscopic, radiometric, electrometric, chromatographic, metabolomic and chemometric investigations of plant products (monomeric species as well as polymeric molecules such as nucleic acids, proteins, lipids and carbohydrates) are included within the remit of the Journal. Papers dealing with novel methods relating to areas such as data handling/ data mining in plant sciences will also be welcomed.