title: Assay for drug discovery: Synthesis and testing of nitrocefin analogues for use as β-lactamase substrates.
abstract: We report on the synthesis of three nitrocefin analogues and their evaluation as substrates for the detection of β-lactamase activity. These compounds are hydrolyzed by all four Ambler classes of β-lactamases. Kinetic parameters were determined with eight different β-lactamases, including VIM-2, NDM-1, KPC-2, and SPM-1. The compounds do not inhibit the growth of clinically important antibiotic-resistant gram-negative bacteria in vitro. These chromogenic compounds have a distinct absorbance spectrum and turn purple when hydrolyzed by β-lactamases. One of these compounds, UW154, is easier to synthesize from commercial starting materials than nitrocefin and should be significantly less expensive to produce.
authors.affiliation: Department of Chemistry, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Ghavami
authors.affiliation: Department of Chemistry, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.
authors.forename: Geneviève
authors.initials: G
authors.lastname: Labbé
authors.affiliation: Chemistry Research Laboratory, University of Oxford, Oxford OX1 3TA, UK.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Brem
authors.affiliation: Department of Chemistry, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.
authors.forename: Valerie J
authors.initials: VJ
authors.lastname: Goodfellow
authors.affiliation: Department of Chemistry, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.
authors.forename: Laura
authors.initials: L
authors.lastname: Marrone
authors.affiliation: Department of Chemistry, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.
authors.forename: Carol A
authors.initials: CA
authors.lastname: Tanner
authors.affiliation: Department of Biochemistry and Molecular Biology and Center for Blood Research, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Dustin T
authors.initials: DT
authors.lastname: King
authors.affiliation: Department of Chemistry, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.
authors.forename: Melinda
authors.initials: M
authors.lastname: Lam
authors.affiliation: Department of Biochemistry and Molecular Biology and Center for Blood Research, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Natalie C J
authors.initials: NC
authors.lastname: Strynadka
authors.affiliation: Departments of Pathology & Laboratory Medicine, Medicine, and Microbiology & Infectious Diseases, University of Calgary, Calgary, Alberta T2L 2K8, Canada.
authors.forename: Dylan R
authors.initials: DR
authors.lastname: Pillai
authors.affiliation: Department of Chemistry and Biochemistry, Laurentian University, Sudbury, Ontario P3E 2C6, Canada.
authors.forename: Stefan
authors.initials: S
authors.lastname: Siemann
authors.affiliation: School of Cellular and Molecular Medicine, University of Bristol, Bristol BS8 1TD, UK.
authors.forename: James
authors.initials: J
authors.lastname: Spencer
authors.affiliation: Chemistry Research Laboratory, University of Oxford, Oxford OX1 3TA, UK.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Schofield
authors.affiliation: Department of Chemistry, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada. Electronic address: dmitrien@uwaterloo.ca.
authors.forename: Gary I
authors.initials: GI
authors.lastname: Dmitrienko
mesh_terms: D055162:Biocatalysis; D002511:Cephalosporins / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D060326:Chemistry Techniques, Synthetic; D004353:Drug Evaluation, Preclinical; D006868:Hydrolysis; D007700:Kinetics; D001618:beta-Lactamases / Q000378:metabolism*
chemical_list: D002511:Cephalosporins; D001618:beta-Lactamases; C021720:nitrocefin
keywords: Antibiotic resistance detection; Chromogenic cephalosporin; Nitrocefin analogues; β-Lactamase