{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,5,26]],"date-time":"2026-05-26T16:07:01Z","timestamp":1779811621490,"version":"3.53.1"},"posted":{"date-parts":[[2025,12,30]]},"group-title":"Microbiology","reference-count":48,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,12,30]],"date-time":"2025-12-30T00:00:00Z","timestamp":1767052800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,22]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  <jats:italic>Salmonella<\/jats:italic>\n                  infections, including typhoid and paratyphoid fevers, impose a substantial burden in low-income countries, contributing to significant morbidity and mortality associated with enteric and systemic infections. Developing effective treatments for\n                  <jats:italic>Salmonella<\/jats:italic>\n                  infections requires further consideration, especially due to widespread antibiotic resistance. Host-targeted therapies, such as AR-12, offer a promising solution to combat drug resistance. AR-12 has demonstrated broad-spectrum antimicrobial activity against various bacterial pathogens, including\n                  <jats:italic>Salmonella enterica<\/jats:italic>\n                  serovar Typhimurium,\n                  <jats:italic>S<\/jats:italic>\n                  . Typhi,\n                  <jats:italic>Francisella tularensis<\/jats:italic>\n                  , and\n                  <jats:italic>F. novicida<\/jats:italic>\n                  , as well as protozoan parasites and fungal pathogens. To expand its HDT potential against\n                  <jats:italic>S. Typhimurium<\/jats:italic>\n                  , we conducted a medicinal chemistry campaign using AR-12 as a scaffold with systematic optimization of various points of diversity and the pyrazole core to develop analogs informed by structure\u2013activity relationship. This work led to the development of 81 AR-12 analogs. Primary screening identified 38 analogs that are both more potent and less cytotoxic than parent compound AR-12, while only three were less potent and more cytotoxic. Further, only seven compounds affected planktonic\n                  <jats:italic>Salmonella<\/jats:italic>\n                  growth below 20 \u00b5M, suggesting host-directed activity in most of the compounds. Twelve analogs were chosen for secondary screening in MDR\n                  <jats:italic>S<\/jats:italic>\n                  . Typhimurium. Compounds 372, 373, and 378 demonstrated remarkable selectivity, with values exceeding 1500 for both susceptible and MDR\n                  <jats:italic>S<\/jats:italic>\n                  . Typhimurium, compared to AR-12\u2019s selectivity of around 20. This approximately 100-fold improvement, coupled with improved potency against intracellular\n                  <jats:italic>Salmonella<\/jats:italic>\n                  , suggests these analogs have significantly greater host-directed activity than direct antibacterial effects. Proteomic analysis for the two most potent compounds, 341 and 370 revealed enrichment of vesicle-mediated transport proteins, specifically with respect to retrograde transport at the trans-Golgi-network and intra-Golgi traffic. These results suggest that the analogs reduce intracellular\n                  <jats:italic>S<\/jats:italic>\n                  . Typhimurium replication by disrupting its exploitation of the host cell\u2019s vesicle-mediated transport system.\n                <\/jats:p>","DOI":"10.64898\/2025.12.30.696991","type":"posted-content","created":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T13:55:46Z","timestamp":1767275746000},"source":"Crossref","is-referenced-by-count":0,"title":["Novel compounds derived from AR-12 that demonstrate host-directed clearance of intracellular\n                  <i>Salmonella enterica<\/i>\n                  Serovar Typhimurium"],"prefix":"10.64898","author":[{"given":"Elizabeth G.","family":"Graham-Gurysh","sequence":"first","affiliation":[{"name":"Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M. Shamim","family":"Hasan Zahid","sequence":"additional","affiliation":[{"name":"Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Devika M.","family":"Varma","sequence":"additional","affiliation":[{"name":"Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antonio","family":"Landavazo","sequence":"additional","affiliation":[{"name":"Center for Drug Discovery, RTI International, Research Triangle Park, Durham, NC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ojas A","family":"Namjoshi","sequence":"additional","affiliation":[{"name":"Center for Drug Discovery, RTI International, Research Triangle Park, Durham, NC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph W","family":"Wilson","sequence":"additional","affiliation":[{"name":"Center for Drug Discovery, RTI International, Research Triangle Park, Durham, NC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monica M.","family":"Johnson","sequence":"additional","affiliation":[{"name":"Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryan N.","family":"Woodring","sequence":"additional","affiliation":[{"name":"Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aaron T.","family":"Hendricksen","sequence":"additional","affiliation":[{"name":"Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph","family":"Vath","sequence":"additional","affiliation":[{"name":"Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erica N.","family":"Pino","sequence":"additional","affiliation":[{"name":"Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eric M.","family":"Bachelder","sequence":"additional","affiliation":[{"name":"Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruce E.","family":"Blough","sequence":"additional","affiliation":[{"name":"Center for Drug Discovery, RTI International, Research Triangle Park, Durham, NC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1820-8382","authenticated-orcid":false,"given":"Kristy M.","family":"Ainslie","sequence":"additional","affiliation":[{"name":"Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA"},{"name":"Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Chapel Hill, NC, USA"},{"name":"Department of Microbiology and Immunology, UNC School of Medicine, University of North Carolina, Chapel Hill, NC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026052608550858000_2025.12.30.696991v2.1","unstructured":"Organization WH. 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