{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T10:01:35Z","timestamp":1785492095477,"version":"3.56.0"},"posted":{"date-parts":[[2026,7,29]]},"group-title":"Biochemistry","reference-count":74,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T00:00:00Z","timestamp":1785283200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"name":"NIH\/NIGMS","award":["GM114611"],"award-info":[{"award-number":["GM114611"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,29]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Tunicamycins are potent inhibitors of dolichyl-phosphate\n                  <jats:italic>N<\/jats:italic>\n                  -acetylglucosamine phosphotransferase (DPAGT1) but are unsuitable for therapeutic development due to non-selective cytotoxicity, acid-labile glycosidic linkages, and poor physicochemical properties. Although prior structural modifications reduced the promiscuous toxicity of tunicamycins, the intrinsic 11\u2032-\u03b2-1\u2033-\u03b1 trehalose-type glycosidic linkage remains chemically unstable, limiting biological durability. Here, we report a rationally engineered scaffold-stabilization strategy in which the acid-labile linkage is replaced with a chemically robust cyclitol framework, enabling the concise synthesis of chemically stable and water-soluble tunicamycin analogues in only 12 synthetic steps. From this platform, TM-Cy-TBPA (\n                  <jats:bold>4<\/jats:bold>\n                  ) was identified as a lead DPAGT1 inhibitor that potently suppresses the proliferation of breast cancer cells by inducing G\n                  <jats:sub>2<\/jats:sub>\n                  -phase arrest followed by apoptosis, while exhibiting minimal cytotoxicity toward nontransformed cells. The compound shows improved solubility, and favorable pharmacokinetic exposure. These results establish tunicamycin cyclitol analogues as a structurally distinct class of selective DPAGT1-targeted anticancer agents and demonstrate that stabilization of the glycosidic linkage is an effective strategy for enhancing pharmacological selectivity, improving\n                  <jats:italic>in vivo<\/jats:italic>\n                  performance, and simplifying the synthetic route.\n                <\/jats:p>","DOI":"10.64898\/2026.07.28.741246","type":"posted-content","created":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T11:10:20Z","timestamp":1785323420000},"source":"Crossref","is-referenced-by-count":0,"title":["Rationally Engineered, Chemically Stable Tunicamycin Analogues Decouple DPAGT1 Inhibition from Non-Selective Toxicity"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6897-8959","authenticated-orcid":false,"given":"Katsuhiko","family":"Mitachi","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Science Center, 881 Madison Avenue, Memphis, Tennessee 38163, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3656-7442","authenticated-orcid":false,"given":"Antonio","family":"S\u00e1nchez-Ruiz","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, Faculty of Sciences, University of M\u00e1laga, Campus de Teatinos 29071 M\u00e1laga, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1745-2586","authenticated-orcid":false,"given":"David","family":"Mingle","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Science Center, 881 Madison Avenue, Memphis, Tennessee 38163, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8606-6710","authenticated-orcid":false,"given":"Iv\u00e1n","family":"Cheng-S\u00e1nchez","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, Faculty of Sciences, University of M\u00e1laga, Campus de Teatinos 29071 M\u00e1laga, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1444-2913","authenticated-orcid":false,"given":"Jacob M.","family":"Kirsh","sequence":"additional","affiliation":[{"name":"Division of Chemistry and Chemical Engineering, California Institute of Technology, 1200 E. California Blvd., Pasadena, California 91125, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5149-3576","authenticated-orcid":false,"given":"Francisco","family":"Sarabia","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, Faculty of Sciences, University of M\u00e1laga, Campus de Teatinos 29071 M\u00e1laga, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0021-889X","authenticated-orcid":false,"suffix":"Jr,","given":"William M.","family":"Clemons","sequence":"additional","affiliation":[{"name":"Division of Chemistry and Chemical Engineering, California Institute of Technology, 1200 E. 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