Santucci, MatteoSpyrakis, FrancescaCross, SimonQuotadamo, AntonioFarina, DavideTondi, DonatellaDe Luca, FilomenaDocquier, Jean-DenisPrieto, Ana IsabelIbacache, ClaudiaBlázquez, JesúsVenturelli, AlbertoCruciani, GabrieleCosti, Maria Paola2023-01-252023-01-252017-12-18http://hdl.handle.net/10668/11925β-Lactamases (BLs) able to hydrolyze β-lactam antibiotics and more importantly the last resort carbapenems, represent a major mechanism of resistance in Gram-negative bacteria showing multi-drug or extensively drug resistant phenotypes. The early detection of BLs responsible of resistant infections is challenging: approaches aiming at the identification of new BLs inhibitors (BLI) can thus serve as the basis for the development of highly needed diagnostic tools. Starting from benzo-[b]-thiophene-2-boronic acid (BZB), a nanomolar inhibitor of AmpC β-lactamase (K i  = 27 nM), we have identified and characterized a set of BZB analogues able to inhibit clinically-relevant β-lactamases, including AmpC, Extended-Spectrum BLs (ESBL), KPC- and OXA-type carbapenemases and metallo-β-lactamases (MBL). A multiligand set of boronic acid (BA) β-lactamase inhibitors was obtained using covalent molecular modeling, synthetic chemistry, enzyme kinetics and antibacterial susceptibility testing. Data confirmed the possibility to discriminate between clinically-relevant β-lactamases on the basis of their inhibition profile. Interestingly, this work also allowed the identification of potent KPC-2 and NDM-1 inhibitors able to potentiate the activity of cefotaxime (CTX) and ceftazidime (CAZ) against resistant clinical isolates (MIC reduction, 32-fold). Our results open the way to the potential use of our set of compounds as a diagnostic tool for the sensitive detection of clinically-relevant β-lactamases.enAttribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/Anti-Bacterial AgentsBacteriaBacterial ProteinsBoronic AcidsCefotaximeCeftazidimeComputational BiologyDrug Resistance, BacterialEnterobacteriaceae InfectionsMicrobial Sensitivity Testsbeta-Lactamase Inhibitorsbeta-Lactamasesresearch article29255163open access10.1038/s41598-017-17399-72045-2322PMC5735191https://www.nature.com/articles/s41598-017-17399-7.pdfhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5735191/pdf