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The covalent addition of l-aminoarabinose to the lipid A moiety of LPS is the main colistin resistance mechanism in the human pathogen Pseudomonas aeruginosa.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objectives<\/jats:title><jats:p>Identification (by in silico screening of a chemical library) of potential inhibitors of ArnT, which catalyses the last committed step of lipid A aminoarabinosylation, and their validation in vitro as colistin adjuvants.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The available ArnT crystal structure was used for a docking-based virtual screening of an in-house library of natural products. The resulting putative ArnT inhibitors were tested in growth inhibition assays using a reference colistin-resistant P. aeruginosa strain. The most promising compound was further characterized for its range of activity, specificity and cytotoxicity. Additionally, the effect of the compound on lipid A aminoarabinosylation was verified by MS analyses of lipid A.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>A putative ArnT inhibitor (BBN149) was discovered by molecular docking and demonstrated to specifically potentiate colistin activity in colistin-resistant P. aeruginosa isolates, without relevant effect on colistin-susceptible strains. BBN149 also showed adjuvant activity against colistin-resistant Klebsiella pneumoniae and low toxicity to bronchial epithelial cells. Lipid A aminoarabinosylation was reduced in BBN149-treated cells, although only partially.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>This study demonstrates that in silico screening targeting ArnT can successfully identify inhibitors of colistin resistance and provides a promising lead compound for the development of colistin adjuvants for the treatment of MDR bacterial infections.<\/jats:p><\/jats:sec>","DOI":"10.1093\/jac\/dkaa200","type":"journal-article","created":{"date-parts":[[2020,4,30]],"date-time":"2020-04-30T11:14:53Z","timestamp":1588245293000},"page":"2564-2572","source":"Crossref","is-referenced-by-count":27,"title":["A novel colistin adjuvant identified by virtual screening for ArnT inhibitors"],"prefix":"10.1093","volume":"75","author":[{"given":"Francesca","family":"Ghirga","sequence":"first","affiliation":[{"name":"Center for Life Nano Science@Sapienza, Italian Institute of Technology, Rome, 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