[en] Metallo-β-lactamases (MBLs) contribute to the resistance of Gram-negative bacteria to carbapenems, last-resort antibiotics at hospital, and MBL inhibitors are urgently needed to preserve these important antibacterial drugs. Here, we describe a series of 1,2,4-triazole-3-thione-based inhibitors displaying an α-amino acid substituent, which amine was mono- or disubstituted by (hetero)aryl groups. Compounds disubstituted by certain nitrogen-containing heterocycles showed submicromolar activities against VIM-type enzymes and strong NDM-1 inhibition (Ki = 10-30 nM). Equilibrium dialysis, native mass spectrometry, isothermal calorimetry (ITC), and X-ray crystallography showed that the compounds inhibited both VIM-2 and NDM-1 at least partially by stripping the catalytic zinc ions. These inhibitors also displayed a very potent synergistic activity with meropenem (16- to 1000-fold minimum inhibitory concentration (MIC) reduction) against VIM-type- and NDM-1-producing ultraresistant clinical isolates, including Enterobacterales and Pseudomonas aeruginosa. Furthermore, selected compounds exhibited no or moderate toxicity toward HeLa cells, favorable absorption, distribution, metabolism, excretion (ADME) properties, and no or modest inhibition of several mammalian metalloenzymes.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Legru, Alice ✱; IBMM, CNRS, Univ Montpellier, ENSCM, 34000 Montpellier, France
Verdirosa, Federica ✱; Dipartimento di Biotecnologie Mediche, Università di Siena, 53100 Siena, Italy
Vo-Hoang, Yen; IBMM, CNRS, Univ Montpellier, ENSCM, 34000 Montpellier, France
Tassone, Giusy ; Dipartimento di Biotecnologie, Chimica e Farmacia, Università di Siena, 53100 Siena, Italy
Vascon, Filippo; Laboratory of Structural Biology, Department of Biology, University of Padua, 35121 Padova, Italy
Thomas, Caitlyn A; Department of Chemistry and Biochemistry, Miami University, Oxford, Ohio 45056, United States
Sannio, Filomena ; Dipartimento di Biotecnologie Mediche, Università di Siena, 53100 Siena, Italy
Corsica, Giuseppina; Dipartimento di Biotecnologie Mediche, Università di Siena, 53100 Siena, Italy
Benvenuti, Manuela ; Dipartimento di Biotecnologie, Chimica e Farmacia, Università di Siena, 53100 Siena, Italy
Feller, Georges ; Université de Liège - ULiège > Département des sciences de la vie > Laboratoire de biochimie
Coulon, Rémi; IBMM, CNRS, Univ Montpellier, ENSCM, 34000 Montpellier, France
Marcoccia, Francesca; Dipartimento di Biotecnologie Mediche, Università di Siena, 53100 Siena, Italy
Devente, Savannah Rowane; Dipartimento di Biotecnologie Mediche, Università di Siena, 53100 Siena, Italy
Bouajila, Ezeddine; IBMM, CNRS, Univ Montpellier, ENSCM, 34000 Montpellier, France
Piveteau, Catherine; Drugs and Molecules for Living System, U1177, Inserm, Université de Lille, Faculté de Pharmacie, 59006 Lille, France
Leroux, Florence; Drugs and Molecules for Living System, U1177, Inserm, Université de Lille, Faculté de Pharmacie, 59006 Lille, France
Deprez-Poulain, Rebecca ; Drugs and Molecules for Living System, U1177, Inserm, Université de Lille, Faculté de Pharmacie, 59006 Lille, France
Deprez, Benoît; Drugs and Molecules for Living System, U1177, Inserm, Université de Lille, Faculté de Pharmacie, 59006 Lille, France
Part of this work was supported by SATT AxLR (RESIST─655, 19/0162). The authors thank Pierre Sanchez for mass spectrometry analyses; Sandrine Warenghem and Alexandre Biela for performing the selectivity profile and the ADME study, respectively, on the ARIADNE platforms of the French national infrastructure ChemBioFrance; and Diamond Light Source for providing us beamtime (BAG proposal MX21741) and the staff of beamline I04 for their assistance with crystal testing and data collection. The authors are grateful to the European Synchrotron Radiation Facility (ESRF, Grenoble, France) and its staff for providing them beamtime and assistance in using beamline ID23–1 (proposal MX-2363). The Dipartimento di Biotecnologie Mediche and the Dipartimento di Biotecnologie, Chimica e Farmacia, University of Siena, were both awarded “Department of Excellence 2018–2022” by the Italian Ministry of Education, University and Research (MIUR, L. 232/2016).
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