{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,8]],"date-time":"2026-03-08T19:17:24Z","timestamp":1772997444143,"version":"3.50.1"},"reference-count":55,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2025,12,13]],"date-time":"2025-12-13T00:00:00Z","timestamp":1765584000000},"content-version":"vor","delay-in-days":12,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,12,1]],"date-time":"2025-12-01T00:00:00Z","timestamp":1764547200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Archiv der Pharmazie"],"published-print":{"date-parts":[[2025,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    The emergence of the severe acute respiratory syndrome coronavirus 2 (SARS\u2010CoV\u20102) has underscored the urgent need for novel antiviral strategies. One of the primary targets of interest is the SARS\u2010CoV\u20102 main protease (Mpro), which plays a crucial role in viral replication. Building on our prior work involving machine learning (ML)\u2010based virtual screening for potential Mpro inhibitors, we sought to experimentally validate top\u2010ranked candidates. Microscale thermophoresis (MST) was used to assess the binding affinity, leading to the identification of three promising hits from a library of 180 compounds. Notably, one compound demonstrated high\u2010affinity binding to SARS\u2010CoV\u20102 Mpro (\n                    <jats:italic>K<\/jats:italic>\n                    <jats:sub>d<\/jats:sub>\n                    \u2009=\u20092.8\u2009\u00b1\u20090.9\u2009\u00b5M). However, enzymatic assays revealed that none of the hit compounds inhibited the activity of the protease, suggesting a non\u2010competitive binding. Docking and molecular dynamics (MD) simulations allowed to identify an accessory site in which the compounds exhibited stable interactions. These findings suggest that the identified compounds may serve as a starting point for the rational design of degradation\u2010inducing strategies, such as proteolysis\u2010targeting chimeras (PROTACs), targeting SARS\u2010CoV\u20102 Mpro, and highlight the value of integrating ML\u2010driven discovery with biophysical and computational validation in antiviral drug development.\n                  <\/jats:p>","DOI":"10.1002\/ardp.70169","type":"journal-article","created":{"date-parts":[[2025,12,13]],"date-time":"2025-12-13T14:01:43Z","timestamp":1765634503000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Support Vector Machine Identification of Small Molecule Binders to an Understudied Allosteric Site of SARS\u2010CoV\u20102 Mpro for Next\u2010Generation PROTAC\u2010Based Therapeutics"],"prefix":"10.1002","volume":"358","author":[{"given":"Enrico Mario Alessandro","family":"Fassi","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Sciences Universit\u00e0 degli Studi di Milano Milano Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3490-1179","authenticated-orcid":false,"given":"Nedra","family":"Mekni","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, Division of Pharmaceutical Chemistry University of Vienna Vienna Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marco","family":"Albani","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences Universit\u00e0 degli Studi di Milano Milano Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sabine","family":"Maehrlein","sequence":"additional","affiliation":[{"name":"Department of Medicinal Chemistry, Institute of Pharmaceutical and Biomedical Sciences Johannes Gutenberg\u2010University Mainz Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Annabelle Carolin","family":"Weldert","sequence":"additional","affiliation":[{"name":"Department of Medicinal Chemistry, Institute of Pharmaceutical and Biomedical Sciences Johannes Gutenberg\u2010University Mainz Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tanja","family":"Schirmeister","sequence":"additional","affiliation":[{"name":"Department of Medicinal Chemistry, Institute of Pharmaceutical and Biomedical Sciences Johannes Gutenberg\u2010University Mainz Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5242-1240","authenticated-orcid":false,"given":"Thierry","family":"Langer","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, Division of Pharmaceutical Chemistry University of Vienna Vienna 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