RESUMO
The bacteria, harboring metallo-ß-lactamases (MßLs), become resistant on most ß-lactam antibiotics, specifically New Delhi metallo-ß-lactamase-1 (NDM-1), which hydrolyzes almost all ß-lactam antibiotics leading to bacterial multiple-drug resistance. It is highly desirable to develop effective NDM-1 inhibitors in reviving the efficacy of existing antibiotics. Here, we report a potent covalently reversible scaffold, 3-Bromopyruvate (3BP) to target the NDM-1 in vitro and in vivo. Enzymatic kinetic studies revealed that 3BP is capable of inhibiting the B1 and B2 MßLs and exhibited the best inhibition on NDM-1 with an IC50 of 2.57 µM, also, it was found to be a dose- and time-dependent inhibitor. The study of inhibition mechanism suggested that 3BP reversibly inactivate NDM-1, and may form a dynamic reversible covalent bond with cysteine at active site of the enzyme. Besides, 3BP effectively restored the activity of five ß-lactam antibiotics on three clinical strains expressing NDM-1, resulting in 2-8-fold reduction in MIC. Moreover, the toxicity evaluation of 3BP against L929 mouse fibroblastic cells indicated that 3BP had low cytotoxicity, implying it may be used as lead molecule for future drug candidate.
Assuntos
Antibacterianos/farmacologia , Piruvatos/farmacologia , beta-Lactamases/metabolismo , Animais , Bactérias/efeitos dos fármacos , Domínio Catalítico/efeitos dos fármacos , Linhagem Celular , Cisteína/metabolismo , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Fibroblastos/microbiologia , Cinética , CamundongosRESUMO
We report the discovery of ebselen-based dual covalent inhibitors of metallo-ß-lactamases. Fluorescence and MALDI-TOF analysis suggested that the scaffold could bind to NDM-1 by forming a S-Se bond with Cys221 and an amide bond with Lys224 of NDM-1, thereby exhibiting selective inhibition and labeling against B1 and B2 subclass enzymes in vitro and in vivo.
