title: 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.
authors.affiliation: Discovery Sciences, AstraZeneca R&D Boston, Waltham, Massachusetts 02451 (USA).
authors.forename: Junhe
authors.initials: J
authors.lastname: Ma
authors.forename: Qing
authors.initials: Q
authors.lastname: Cao
authors.forename: Sarah M
authors.initials: SM
authors.lastname: McLeod
authors.forename: Keith
authors.initials: K
authors.lastname: Ferguson
authors.forename: Ning
authors.initials: N
authors.lastname: Gao
authors.forename: Alexander L
authors.initials: AL
authors.lastname: Breeze
authors.forename: Jun
authors.initials: J
authors.lastname: Hu
mesh_terms: D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D000077731:Meropenem; D011485:Protein Binding; D066244:Proton Magnetic Resonance Spectroscopy; D013845:Thienamycins / Q000737:chemistry / Q000378:metabolism; D001618:beta-Lactamases / Q000737:chemistry* / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D013845:Thienamycins; D001618:beta-Lactamases; C552864:beta-lactamase NDM-1; D000077731:Meropenem
keywords: NMR spectroscopy; New Delhi metallo-β-lactamase; drug discovery; in vivo methods; whole-cell screening