Target‐Based Whole‐Cell Screening by ¹H NMR Spectroscopy

Abstract

An NMR-based approach marries the two traditional screening technologies (phenotypic and target-based screening) to find compounds inhibiting a specific enzymatic reaction in bacterial cells. Building on a previous study in which it was demonstrated that hydrolytic decomposition of meropenem in living Escherichia coli cells carrying New Delhi metallo-β-lactamase subclass 1 (NDM-1) can be monitored in real time by NMR spectroscopy, we designed a cell-based NMR screening platform. A strong NDM-1 inhibitor was identified with cellular IC50 of 0.51 μm, which is over 300-fold more potent than captopril, a known NDM-1 inhibitor. This new screening approach has great potential to be applied to targets in other cell types, such as mammalian cells, and to targets that are only stable or functionally competent in the cellular environment.

N-Aryl mercaptoacetamides as potential multi-target inhibitors of metallo-β-lactamases (MBLs) and the virulence factor LasB from Pseudomonas aeruginosa

“…Substantial synthetic work has already been done on this scaffold to obtain novel MBL inhibitors, and co-crystallization experiments have been reported. 42–46 …”

Structural Basis of Metallo-β-Lactamase Inhibition by Captopril Stereoisomers

“…l -Captopril was used successfully for several decades to control high blood pressure. The clinically used (2 S ,2 S )-stereoisomer, i.e., l -captopril, inhibits several MBLs from all subclasses ( 14 – 18 ). However, the (2 S ,2 R )-stereoisomer, (2 S )-1-[(2 R )-2-methyl-3-sulfanylpropanoyl] pyrrolidine-2-carboxylic acid ( Fig.…”

Discovery of thiosemicarbazone derivatives as effective New Delhi metallo-β-lactamase-1 (NDM-1) inhibitors against NDM-1 producing clinical isolates

“…The majority of reported inhibitors of NDM-1 were non-covalent inhibitors 12 , including chelating agents and zinc coordinating agents, such as compounds 1 – 4 13 , 14 , 15 , 16 ( Fig. 1 ).…”

“…Compound 1 was an inhibitor of NDM-1 and other MBLs as well, whereas it was completely ineffective against the metallo- β -lactamases class A, C and D 16 . Compounds 3 and 4 (methisazone) were zinc coordinating inhibitors, molecules able to coordinate the zinc ions within the NDM-1 binding site 13 , 15 . Although some effective molecules have been reported, concurrently, there are neither clinical drugs which can reverse the resistance mediated by NDM-1 nor approved NDM-1 inhibitors which are in clinical trials.…”