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The Use of a β-lactamase-based Conductimetric Biosensor Assay to Detect Biomolecular Interactions
Published on: February 1, 2018
Amino Acid Thioesters Exhibit Inhibitory Activity against B1-B3 Subclasses of Metallo-β-lactamases
Yue-Juan Zhang1, Xiao-Long Liu1, Wen-Ming Wang1
1Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, Chemical Biology Innovation Laboratory, College of Chemistry and Materials Science, Northwest University.
Abstract:
Superbug infection caused by metallo-β-lactamases (MβLs) is a global public health threat. Previous studies reported that the thioesters specifically inhibited the B3 subclass MβL L1. In this work, nine amino acid thioesters 1-9 were synthesized, the activity evaluation revealed that all of these molecules exhibited broad-spectrum inhibitory efficacy against ImiS, IMP-1, NDM-1, and L1, with IC50 values range of 0.02-54.9 µM (except 5 and 7 on NDM-1), and 1 was found to be the best inhibitor with IC50 range of 0.02-16.63 µM. Minimal inhibitory concentration (MIC) assays showed that thioesters 1, 5 and 9 restored 2-32-fold antibacterial activity of cefazolin and/or imipenem against both Escherichia coli BL21 and DH10B strain expressing ImiS, L1, IMP-1 and NDM-1 (except 5 on NDM-1), and also, thioester 1 increased 2-4-fold antimicrobial activity of cefazolin on two clinical strains Pseudomonas aeruginosa and Klebsiella pneumoniae producing NDM-1. Stability evaluation indicated that thioester 1 was partially hydrolysed by MβLs to be converted into the mercaptoacetic acid, revealing that the thioester and its hydrolysate mercaptoacetic acid jointly inhibit MβLs. Isothermal titration calorimetry (ITC) monitoring showed that thioester 1 exhibited dose-dependent inhibition on four MβLs tested, and the binding of 1/L1 showed mainly enthalpy driven, while 1/NDM-1 was found to be more entropy driven. Docking studies suggested that 1 bound to Zn(II) ion(s) preferentially via its carboxylate group, while other moieties interacted mostly with the conserved active site residues.
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