title: Rhodanine hydrolysis leads to potent thioenolate mediated metallo-β-lactamase inhibition.
abstract: The use of β-lactam antibiotics is compromised by resistance, which is provided by β-lactamases belonging to both metallo (MBL)- and serine (SBL)-β-lactamase subfamilies. The rhodanines are one of very few compound classes that inhibit penicillin-binding proteins (PBPs), SBLs and, as recently reported, MBLs. Here, we describe crystallographic analyses of the mechanism of inhibition of the clinically relevant VIM-2 MBL by a rhodanine, which reveal that the rhodanine ring undergoes hydrolysis to give a thioenolate. The thioenolate is found to bind via di-zinc chelation, mimicking the binding of intermediates in β-lactam hydrolysis. Crystallization of VIM-2 in the presence of the intact rhodanine led to observation of a ternary complex of MBL, a thioenolate fragment and rhodanine. The crystallographic observations are supported by kinetic and biophysical studies, including (19)F NMR analyses, which reveal the rhodanine-derived thioenolate to be a potent broad-spectrum MBL inhibitor and a lead structure for the development of new types of clinically useful MBL inhibitors.
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford OX1 3TA, UK.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Brem
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford OX1 3TA, UK.
authors.forename: Sander S
authors.initials: SS
authors.lastname: van Berkel
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford OX1 3TA, UK.
authors.forename: WeiShen
authors.initials: W
authors.lastname: Aik
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford OX1 3TA, UK.
authors.forename: Anna M
authors.initials: AM
authors.lastname: Rydzik
authors.affiliation: School of Cellular and Molecular Medicine, University of Bristol, Medical Sciences Building, Bristol BS8 1TD, UK.
authors.forename: Matthew B
authors.initials: MB
authors.lastname: Avison
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford OX1 3TA, UK.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Pettinati
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford OX1 3TA, UK.
authors.forename: Klaus-Daniel
authors.initials: KD
authors.lastname: Umland
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford OX1 3TA, UK.
authors.forename: Akane
authors.initials: A
authors.lastname: Kawamura
authors.affiliation: School of Cellular and Molecular Medicine, University of Bristol, Medical Sciences Building, Bristol BS8 1TD, UK.
authors.forename: James
authors.initials: J
authors.lastname: Spencer
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford OX1 3TA, UK.
authors.forename: Timothy D W
authors.initials: TD
authors.lastname: Claridge
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford OX1 3TA, UK.
authors.forename: Michael A
authors.initials: MA
authors.lastname: McDonough
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford OX1 3TA, UK.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Schofield
mesh_terms: D001703:Biophysics; D003461:Crystallography; D006868:Hydrolysis; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D000077731:Meropenem; D012236:Rhodanine / Q000737:chemistry* / Q000494:pharmacology; D013845:Thienamycins / Q000494:pharmacology; D065093:beta-Lactamase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D001618:beta-Lactamases / Q000737:chemistry
chemical_list: D013845:Thienamycins; D065093:beta-Lactamase Inhibitors; D012236:Rhodanine; D001618:beta-Lactamases; D000077731:Meropenem