20.500.12556/RUL-141439 Covalent inhibitors of bacterial peptidoglycan biosynthesis enzyme MurA with chloroacetamide warhead MurA (UDP-N-acetylglucosamine enolpyruvyl transferase) catalyzes the first committed step in the cytoplasmic part of peptidoglycan biosynthesis and is a validated target enzyme for antibacterial drug discovery; the inhibitor fosfomycin has been used clinically for decades. Like fosfomycin, most MurA inhibitors are small heterocyclic compounds that inhibit the enzyme by forming a covalent bond with the active site cysteine. The reactive chloroacetamide group was selected from a series of suitable electrophilic thiol-reactive warheads. The predominantly one-step synthesis led to the construction of the final library of 47 fragment-sized chloroacetamide compounds. Several new E. coli MurA inhibitors were identified, with the most potent compound having an IC$_{50}$ value in the low micromolar range. The electrophilic reactivity of all chloroacetamide fragments in our library was evaluated by a high-throughput spectrophotometric assay using the reduced Ellman reagent as a surrogate for the cysteine thiol. LC-MS/MS experiments confirmed the covalent binding of the most potent inhibitor to Cys115 of the digested MurA enzyme. The covalent binding was further investigated by a biochemical time-dependent assay and a dilution assay, which confirmed the irreversible and time-dependent mode of action. The efficacy of chloroacetamide derivatives against MurA does not correlate with their thiol reactivity, making the active fragments valuable starting points for fragment-based development of new antibacterial agents targeting MurA. antibacterial agents UDP-N-acetylglucosamine enolpyruvyl transferase thiol reactivity antibakterijska sredstva UDP-N-acetilglukozamin enolpiruvil transferaza tiolna reaktivnost bakterije farmacevtska kemija true false true Angleški jezik Slovenski jezik Članek v reviji 2022-09-29 10:12:24 2022-09-29 10:12:28 2023-01-20 09:18:46 0000-00-00 00:00:00 2022 0 0 15 str. Vol. 243, art. 114752 2022 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 579.8:615.4:54 0223-5234 10.1016/j.ejmech.2022.114752 123442179 1-s2.0-S0223523422006547-main.pdf 1-s2.0-S0223523422006547-main.pdf 1 EF948C680A1D00A7A5A8DE1234711C08 f783853c8712e9cc5357e46896c29a6727a28a7ed7ec481c1d334bb322017d0e a2d2ab62-3fce-11ed-9c94-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=162497 https://www.sciencedirect.com/science/article/pii/S0223523422006547 0 733beb8b-3fce-11ed-9c94-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=162496 Fakulteta za farmacijo Veterinarska fakulteta 0 0 0