Katarina Grabrijan (Author), Martina Hrast (Author), Matic Proj (Author), Ana Dolšak (Author), Irena Zdovc (Author), Tímea Imre (Author), László Petri (Author), Péter Ábrányi-Balogh (Author), György Keserü M. (Author), Stanislav Gobec (Author)

Abstract

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 IC50 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.

Keywords

antibakterijska sredstva;UDP-N-acetilglukozamin enolpiruvil transferaza;tiolna reaktivnost;antibacterial agents;UDP-N-Acetylglucosamine enolpyruvyl transferase;thiol reactivity;

Data

Language: English
Year of publishing:
Typology: 1.01 - Original Scientific Article
Organization: UL FFA - Faculty of Pharmacy
UDC: 579.8:615.4:54
COBISS: 123442179 Link will open in a new window
ISSN: 0223-5234
Views: 77
Downloads: 28
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Other data

Secondary language: Slovenian
Secondary keywords: Bakterije;Farmacevtska kemija;
Type (COBISS): Article
Pages: 15 str.
Issue: ǂVol. ǂ243
Chronology: 2022
DOI: 10.1016/j.ejmech.2022.114752
ID: 16608099