{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,10]],"date-time":"2026-04-10T23:57:53Z","timestamp":1775865473659,"version":"3.50.1"},"reference-count":42,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2024,11,20]],"date-time":"2024-11-20T00:00:00Z","timestamp":1732060800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Phytochemical Analysis"],"published-print":{"date-parts":[[2025,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:sec><jats:title>Introduction<\/jats:title><jats:p>Drug discovery research targeting SARS\u2010CoV\u20102 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\u2010MS analysis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objective<\/jats:title><jats:p>The aim of this study is to construct a biolayer interferometry (BLI)\u2010based system for recognizing active ingredients that inhibit the main protease (Mpro) of SARS\u2010CoV\u20102 and to identify the active chemical components binding to Mpro from herbal medicines.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methodology<\/jats:title><jats:p>We developed a novel FRET fluorogenic probe by linking the amino acid sequences of the fluorescent proteins Lssmorange and mKate2 (Ls\u2010mK). The interaction between traditional Chinese medicine and Mpro was analyzed using BLI. Ultrahigh performance liquid chromatography coupled with quadrupole time\u2010of\u2010flight mass spectrometry (UPLC\u2010QTOF\u2010MS) was employed to analyze the composition of herbal medicines.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Fluorescence detection and spectroscopy confirmed the successful construction of an Mpro inhibitor screening system. Lanqin Oral Liquid (LQL) and <jats:italic>Gardeniae fructus<\/jats:italic> exhibited strong inhibitory effects on Mpro. Ten compounds were identified from <jats:italic>G. fructus<\/jats:italic> 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\u2009\u03bcM and 801\u2009nM, respectively. The half\u2010maximal inhibitory concentrations (IC50) of DAAME and Gardoside for Mpro were 27.46 and 13.7\u2009\u03bcM, respectively.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>This study established a functional Mpro inhibitor screening system. Among the 10 components identified from <jats:italic>G. fructus<\/jats:italic> that bind to Mpro, DAAME and Gardoside displayed strong binding and inhibitory activity, indicating their potential as lead compounds for inhibiting SARS\u2010CoV\u20102 viral replication.<\/jats:p><\/jats:sec>","DOI":"10.1002\/pca.3462","type":"journal-article","created":{"date-parts":[[2024,11,21]],"date-time":"2024-11-21T01:46:43Z","timestamp":1732153603000},"page":"718-731","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["A Biolayer Interferometry\u2010Based SARS\u2010COV\u20102 Mpro\u2010Targeted Active Ingredients Recognition System: Construction and Application in Ligand Screening From Herbal Medicines"],"prefix":"10.1002","volume":"36","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3010-5965","authenticated-orcid":false,"given":"Dai","family":"Zhang","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine First Affiliated Hospital of Henan University of Traditional Chinese Medicine  Zhengzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bing","family":"Han","sequence":"additional","affiliation":[{"name":"First Affiliated Hospital Henan Provincial Center for Clinical Application, Evaluation, and Transformation of Traditional Chinese Medicine Henan Provincial Center for Safety Evaluation and Risk Control of Traditional Chinese Medicine Henan Provincial Key Laboratory of Traditional Chinese Medicine Clinical Pharmacy Henan University of Traditional Chinese Medicine  Zhengzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiao\u2010Fei","family":"Chen","sequence":"additional","affiliation":[{"name":"First Affiliated Hospital Henan Provincial Center for Clinical Application, Evaluation, and Transformation of Traditional Chinese Medicine Henan Provincial Center for Safety Evaluation and Risk Control of Traditional Chinese Medicine Henan Provincial Key Laboratory of Traditional Chinese Medicine Clinical Pharmacy Henan University of Traditional Chinese Medicine  Zhengzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shuai","family":"Zhao","sequence":"additional","affiliation":[{"name":"Henan Provincial Key Laboratory of Pediatric Genes and Metabolic Diseases Zhengzhou University Affiliated Children's Hospital  Zhengzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wei\u2010Xia","family":"Li","sequence":"additional","affiliation":[{"name":"First Affiliated Hospital Henan Provincial Center for Clinical Application, Evaluation, and Transformation of Traditional Chinese Medicine Henan Provincial Center for Safety Evaluation and Risk Control of Traditional Chinese Medicine Henan Provincial Key Laboratory of Traditional Chinese Medicine Clinical Pharmacy Henan University of Traditional Chinese Medicine  Zhengzhou China"},{"name":"Department of Pharmacy First Affiliated Hospital of Henan University of Chinese Medicine  Zhengzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hui","family":"Zhang","sequence":"additional","affiliation":[{"name":"First Affiliated Hospital Henan Provincial Center for Clinical Application, Evaluation, and Transformation of Traditional Chinese Medicine Henan Provincial Center for Safety Evaluation and Risk Control of Traditional Chinese Medicine Henan Provincial Key Laboratory of Traditional Chinese Medicine Clinical Pharmacy Henan University of Traditional Chinese Medicine  Zhengzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ming\u2010Liang","family":"Zhang","sequence":"additional","affiliation":[{"name":"First Affiliated Hospital Henan Provincial Center for Clinical Application, Evaluation, and Transformation of Traditional Chinese Medicine Henan Provincial Center for Safety Evaluation and Risk Control of Traditional Chinese Medicine Henan Provincial Key Laboratory of Traditional Chinese Medicine Clinical Pharmacy Henan University of Traditional Chinese Medicine  Zhengzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meng\u2010Qi","family":"Huo","sequence":"additional","affiliation":[{"name":"First Affiliated Hospital Henan Provincial Center for Clinical Application, Evaluation, and Transformation of Traditional Chinese Medicine Henan Provincial Center for Safety Evaluation and Risk Control of Traditional Chinese Medicine Henan Provincial Key Laboratory of Traditional Chinese Medicine Clinical Pharmacy Henan University of Traditional Chinese Medicine  Zhengzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yong\u2010Sheng","family":"Qiu","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology Zhengzhou University Affiliated Children's Hospital  Zhengzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ying\u2010Jie","family":"Ren","sequence":"additional","affiliation":[{"name":"First Affiliated Hospital Henan Provincial Center for Clinical Application, Evaluation, and Transformation of Traditional Chinese Medicine Henan Provincial Center for Safety Evaluation and Risk Control of Traditional Chinese Medicine Henan Provincial Key Laboratory of Traditional Chinese Medicine Clinical Pharmacy Henan University of Traditional Chinese Medicine  Zhengzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yao\u2010Dong","family":"Zhang","sequence":"additional","affiliation":[{"name":"Henan Provincial Key Laboratory of Pediatric Genes and Metabolic Diseases Zhengzhou University Affiliated Children's Hospital  Zhengzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xian\u2010Qing","family":"Ren","sequence":"additional","affiliation":[{"name":"Department of Pediatrics Affiliated Hospital of Henan University of Chinese Medicine  Zhengzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wei","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Infection First Affiliated Hospital of Henan University of Traditional Chinese Medicine  Zhengzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jin\u2010Fa","family":"Tang","sequence":"additional","affiliation":[{"name":"First Affiliated Hospital Henan Provincial Center for Clinical Application, Evaluation, and Transformation of Traditional Chinese Medicine Henan Provincial Center for Safety Evaluation and Risk Control of Traditional Chinese Medicine Henan Provincial Key Laboratory of Traditional Chinese Medicine Clinical Pharmacy Henan University of Traditional Chinese Medicine  Zhengzhou China"},{"name":"Department of Pharmacy First Affiliated Hospital of Henan University of Chinese Medicine  Zhengzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,11,20]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2012-7"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2001017"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(22)00152-0"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41577-021-00662-4"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12879-022-07418-y"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.09.018"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2223-y"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41422-021-00581-y"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.abb4489"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.2174\/1389557520666200807134921"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.antiviral.2015.12.012"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1021\/np200906s"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jep.2021.113869"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1002\/ptr.7008"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2091-15-19"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1002\/dta.2123"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jep.2015.02.017"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1039\/D2NP00080F"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11101-014-9335-7"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-021-03170-5"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-016-9633-6"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.9b05479"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.apsb.2021.08.030"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.3390\/molecules28207053"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1186\/s40001-023-01006-9"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1002\/jssc.202100360"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13020-017-0155-8"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-18233-x"},{"issue":"9","key":"e_1_2_8_30_1","first-page":"3428","article-title":"Identification of New Anti\u2010nCoV Drug Chemical Compounds From Indian Spices Exploiting SARS\u2010CoV\u20102 Main Protease as Target","volume":"39","author":"Kundu D.","year":"2021","journal-title":"Journal of Biomolecular Structure & Dynamics"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pharmthera.2021.107843"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1875-5364(20)60031-0"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.phymed.2020.153279"},{"issue":"9","key":"e_1_2_8_34_1","first-page":"3347","article-title":"An Investigation Into the Identification of Potential Inhibitors of SARS\u2010CoV\u20102 Main Protease Using Molecular Docking Study","volume":"39","author":"Das S.","year":"2021","journal-title":"Journal of Biomolecular Structure & Dynamics"},{"issue":"1","key":"e_1_2_8_35_1","first-page":"17","article-title":"High\u2010Throughput Screening of SARS\u2010CoV\u20102 Main and Papain\u2010Like Protease Inhibitors","volume":"14","author":"Zang Y.","year":"2023","journal-title":"Protein & Cell"},{"key":"e_1_2_8_36_1","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2023.1193282"},{"key":"e_1_2_8_37_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12951-021-00965-7"},{"key":"e_1_2_8_38_1","doi-asserted-by":"publisher","DOI":"10.1002\/smll.202303059"},{"key":"e_1_2_8_39_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41401-020-0483-6"},{"key":"e_1_2_8_40_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells10102600"},{"key":"e_1_2_8_41_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jep.2023.117397"},{"key":"e_1_2_8_42_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jfda.2016.11.005"},{"key":"e_1_2_8_43_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11418-022-01627-0"}],"container-title":["Phytochemical Analysis"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pca.3462","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,21]],"date-time":"2025-10-21T09:31:24Z","timestamp":1761039084000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/pca.3462"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,11,20]]},"references-count":42,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2025,4]]}},"alternative-id":["10.1002\/pca.3462"],"URL":"https:\/\/doi.org\/10.1002\/pca.3462","archive":["Portico"],"relation":{},"ISSN":["0958-0344","1099-1565"],"issn-type":[{"value":"0958-0344","type":"print"},{"value":"1099-1565","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,11,20]]},"assertion":[{"value":"2024-09-10","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-10-05","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-11-20","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}