{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,13]],"date-time":"2026-04-13T14:45:53Z","timestamp":1776091553211,"version":"3.50.1"},"reference-count":36,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,12,18]],"date-time":"2024-12-18T00:00:00Z","timestamp":1734480000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>The unavoidable propagation of the coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has underscored the urgent requirement for efficacious therapeutic agents. The dried fruit of <jats:italic>Terminalia chebula<\/jats:italic> Retz., namely Chebulae Fructus, is widely used for treating bacterial and viral infectious diseases, which was witnessed to perform anti-SARS-CoV-2 activity in recommended Chinese patent medicine.<\/jats:p><\/jats:sec><jats:sec><jats:title>Aim<\/jats:title><jats:p>SARS-CoV-2 main protease (M<jats:sup>pro<\/jats:sup>) and papain-like protease (PL<jats:sup>pro<\/jats:sup>) present essential effects on SARS-CoV-2 replication and transcription, considering as the attractive targets for therapeutic intervention. In this study, we focused on the dual-target to obtain broad-spectrum antiviral candidates from Chebulae Fructus.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The identified compounds from Chebulae Fructus were used to build a library in a previous study, which were evaluated by molecular docking to screen potential antiviral agents. The SARS-CoV-2 M<jats:sup>pro<\/jats:sup> and PL<jats:sup>pro<\/jats:sup> were expressed in <jats:italic>E. coli<\/jats:italic> cells and purified. Fluorescence resonance energy transfer (FRET) and surface plasmon resonance (SPR) were utilized to verify the affinity with dual targets. SARS-CoV-2 wild-type, Omicron BA.5 and Omicron EG.5 variants were employed to validate their antiviral activities <jats:italic>in vitro<\/jats:italic>. Molecular dynamics simulation was conducted via Gromacs 2022 software in 500\u202fns to unveil the conformation stability.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Targeting on M<jats:sup>pro<\/jats:sup> and PL<jats:sup>pro<\/jats:sup>, eight compounds were screened as the potential dual-target inhibitors in molecular docking. In FRET and SPR assays, 1,2,3,4,6-penta-<jats:italic>O<\/jats:italic>-galloyl-<jats:italic>\u03b2<\/jats:italic>-D-glucose (PGG) and 1,2,3,6-tetra-<jats:italic>O<\/jats:italic>-galloyl-<jats:italic>\u03b2<\/jats:italic>-D-glucose (TGG) showed good inhibitory activities with IC<jats:sub>50<\/jats:sub> values ranging from 1.33 to 27.37\u202f\u03bcM, and affinity with <jats:italic>K<jats:sub>D<\/jats:sub><\/jats:italic> values ranging from 0.442 to 0.776\u202f\u03bcM. Satisfactorily, both PGG and TGG display antiviral activity <jats:italic>in vitro<\/jats:italic> with EC<jats:sub>50<\/jats:sub> values ranging from 3.20 to 37.29\u202f\u03bcM, suggesting as the promising candidates against SARS-CoV-2. In molecular dynamics simulation study, the complexes of M<jats:sup>pro<\/jats:sup>-PGG, M<jats:sup>pro<\/jats:sup>-TGG, PL<jats:sup>pro<\/jats:sup>-PGG, and PL<jats:sup>pro<\/jats:sup>-TGG exhibited stability over 500\u202fns period, unveiling the potential interactions.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>PGG and TGG are the promising dual-target inhibitors of SARS-CoV-2, which may avoid drug resistance and have a good development prospect. The outcomes of this study provide an effective strategy to systematically explore the antiviral bioactive compounds from Chebulae Fructus.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fmicb.2024.1510665","type":"journal-article","created":{"date-parts":[[2024,12,18]],"date-time":"2024-12-18T14:54:28Z","timestamp":1734533668000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["High-throughput screening of dual-target inhibitors for SARS-CoV-2 main protease and papain-like protease from Chebulae Fructus: in silico prediction and experimental verification"],"prefix":"10.3389","volume":"15","author":[{"given":"Changjian","family":"Wang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yipeng","family":"Cao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qi","family":"Yang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xinyue","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhiying","family":"Yang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jingjing","family":"Yang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xinru","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bin","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuefei","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Min","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2024,12,18]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"457","DOI":"10.1055\/s-2002-32070","article-title":"Inhibition of HIV-1 integrase by galloyl glucoses from Terminalia chebula and flavonol glycoside gallates from Euphorbia pekinensis","volume":"68","author":"Ahn","year":"2002","journal-title":"Planta Med."},{"key":"ref2","doi-asserted-by":"publisher","first-page":"1813","DOI":"10.1056\/NEJMoa2007764","article-title":"Remdesivir for the treatment of COVID-19-final report","volume":"383","author":"Beigel","year":"2020","journal-title":"N. 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