{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T05:47:00Z","timestamp":1782280020216,"version":"3.54.5"},"posted":{"date-parts":[[2025,1,31]]},"group-title":"Biochemistry","reference-count":53,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,1,31]],"date-time":"2025-01-31T00:00:00Z","timestamp":1738281600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,1,31]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  MraY, a bacterial enzyme crucial for the synthesis of peptidoglycans, represents a promising yet underexplored target for the development of effective antibacterial agents. Nature has provided several classes of nucleoside inhibitors of MraY and scientists have modified these structures further to obtain natural product-like inhibitors of MraY. The natural products and their synthetic analogs suffer from non-optimal\n                  <jats:italic>in vivo<\/jats:italic>\n                  efficacy, and the synthetic complexity of the structures renders the synthesis and structure-activity relationship (SAR) studies of these molecules particularly challenging. In this study, we present our findings on the discovery of first-in-class 1,2,4-triazole-based MraY inhibitors that are not nucleoside-derived. A series of 1,2,4-triazole analogous were identified by a structure\u2013activity-relationship (SAR) study using a structure-based drug design strategy. Compound\n                  <jats:bold>1<\/jats:bold>\n                  , with an IC\n                  <jats:sub>50<\/jats:sub>\n                  of 171 \u00b5M against MraY from\n                  <jats:italic>Staphylococcus aureus<\/jats:italic>\n                  (MraY\n                  <jats:italic>\n                    <jats:sub>SA<\/jats:sub>\n                  <\/jats:italic>\n                  ), was optimized to compound\n                  <jats:bold>12a<\/jats:bold>\n                  , exhibiting an IC\n                  <jats:sub>50<\/jats:sub>\n                  of 25 \u00b5M. Molecular docking studies against MraY\n                  <jats:italic>\n                    <jats:sub>SA<\/jats:sub>\n                  <\/jats:italic>\n                  provided insights into these compounds\u2019 binding interactions and activity. Furthermore, screening against the ESKAPE bacterial panel was also conducted, through which we discovered compounds demonstrating broad-spectrum antibacterial activity against\n                  <jats:italic>E. faecium<\/jats:italic>\n                  , methicillin-resistant\n                  <jats:italic>S. aureus<\/jats:italic>\n                  (MRSA), vancomycin-resistant Enterococci (VRE) strains and\n                  <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                  . The novel, first-in-class non-nucleoside inhibitors of MraY highlighted in this work provide a strong proof-of-concept of how to leverage structural information of the protein to develop future antibacterial agents targeting MraY.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Abstract Figure<\/jats:title>\n                  <jats:fig id=\"ufig1\" position=\"float\" orientation=\"portrait\" fig-type=\"figure\">\n                    <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"635793v1_ufig1\" position=\"float\" orientation=\"portrait\"\/>\n                  <\/jats:fig>\n                <\/jats:sec>","DOI":"10.1101\/2025.01.30.635793","type":"posted-content","created":{"date-parts":[[2025,1,31]],"date-time":"2025-01-31T22:25:15Z","timestamp":1738362315000},"source":"Crossref","is-referenced-by-count":0,"title":["1,2,4-Triazole-based first-in-class non-nucleoside inhibitors of bacterial enzyme MraY"],"prefix":"10.64898","author":[{"given":"Tomayo","family":"Berida","sequence":"first","affiliation":[{"name":"Department of BioMolecular Sciences, University of Mississippi, University, MS, 38677, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tzu-Yu","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of BioMolecular Sciences, University of Mississippi, University, MS, 38677, USA"},{"name":"Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, Memphis, TN, 38103, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefanie C.","family":"Weck","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Pharmaceutical and Medicinal Chemistry, Saarland University, Saarbr\u00fccken, 66123, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcel","family":"Lutz","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Pharmaceutical and Medicinal Chemistry, Saarland University, Saarbr\u00fccken, 66123, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samuel R.","family":"McKee","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology, Center for Women\u2019s Infectious Disease Research, Washington University School of Medicine, St. Louis, MO, 63110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nathalie","family":"Kagerah","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Pharmaceutical and Medicinal Chemistry, Saarland University, Saarbr\u00fccken, 66123, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Destinee","family":"Manning","sequence":"additional","affiliation":[{"name":"Department of BioMolecular Sciences, University of Mississippi, University, MS, 38677, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohamed E.","family":"Jahan","sequence":"additional","affiliation":[{"name":"Department of BioMolecular Sciences, University of Mississippi, University, MS, 38677, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3080-9754","authenticated-orcid":false,"given":"Sushil K.","family":"Mishra","sequence":"additional","affiliation":[{"name":"Department of BioMolecular Sciences, University of Mississippi, University, MS, 38677, USA"},{"name":"Department of Biomedical Engineering, University of Mississippi, University, MS, 38677, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert J.","family":"Doerksen","sequence":"additional","affiliation":[{"name":"Department of BioMolecular Sciences, University of Mississippi, University, MS, 38677, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christina L.","family":"Stallings","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology, 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