title: Novel non-β-lactam inhibitor of β-lactamase TEM-171 based on acylated phenoxyaniline.
abstract: The microbial resistance to antibiotics is a genuine global threat. Consequently, a search of new inhibitors remains of acute importance due to the increasing spread of multidrug resistance. Here we present a new type of non-β-lactam β-lactamase inhibitor PA-34 based on natural phenoxyaniline, identified using computer-assisted screening of scaffolds related to those of known low-affinity inhibitors. The compound displays reversible competitive inhibition of bacterial β-lactamase TEM-171, with a Kiof 88 μM. Using enzyme kinetics, infra-red spectroscopy, fluorescence quenching and computer docking, we propose that the inhibitor binds at the entrance to the enzyme active site. This is a novel inhibition mechanism compared to binding covalently to the catalytic serine in the active site or non-covalently to the allosteric site. The residues involved in binding the inhibitor are conserved among molecular class A β-lactamases. The identified compound and its proposed binding mode may have a potential for a regulation of the catalytic activity of a wide range of class A β-lactamases. We also hypothesise that the presented route for finding non-β-lactam compounds may be an effective and durable approach for combating bacterial antibiotic resistance.
authors: Chemistry Department, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia. Electronic address: vitaly.grigorenko@gmail.com.
authors: V G
authors: VG
authors: Grigorenko
authors: Chemistry Department, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia.
authors: I P
authors: IP
authors: Andreeva
authors: Chemistry Department, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia.
authors: M Yu
authors: MY
authors: Rubtsova
authors: Chemistry Department, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia.
authors: I M
authors: IM
authors: Deygen
authors: Chemistry Department, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia.
authors: R L
authors: RL
authors: Antipin
authors: Chemistry Department, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia.
authors: A G
authors: AG
authors: Majouga
authors: Chemistry Department, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia.
authors: A M
authors: AM
authors: Egorov
authors: European Molecular Biology Laboratory (EMBL), c/o DESY, Notkestrasse 85, Building 25A, 22607 Hamburg, Germany.
authors: D A
authors: DA
authors: Beshnova
authors: European Molecular Biology Laboratory (EMBL), c/o DESY, Notkestrasse 85, Building 25A, 22607 Hamburg, Germany.
authors: J
authors: J
authors: Kallio
authors: European Molecular Biology Laboratory (EMBL), c/o DESY, Notkestrasse 85, Building 25A, 22607 Hamburg, Germany.
authors: C
authors: C
authors: Hackenberg
authors: European Molecular Biology Laboratory (EMBL), c/o DESY, Notkestrasse 85, Building 25A, 22607 Hamburg, Germany.
authors: V S
authors: VS
authors: Lamzin
mesh_terms: D000215:Acylation; D000814:Aniline Compounds / Q000737:chemistry / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D020134:Catalytic Domain; D003198:Computer Simulation; D055808:Drug Discovery / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D005453:Fluorescence; D007700:Kinetics; D062105:Molecular Docking Simulation; D015394:Molecular Structure; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods; D065093:beta-Lactamase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D001618:beta-Lactamases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000814:Aniline Compounds; D001426:Bacterial Proteins; D065093:beta-Lactamase Inhibitors; D001618:beta-Lactamases
keywords: Antibiotic resistance; Lead discovery; Non-β-lactam inhibitor; Recombinant β-lactamase TEM