title: Kinetics of avibactam inhibition against Class A, C, and D β-lactamases.
abstract: Avibactam is a non-β-lactam β-lactamase inhibitor with a spectrum of activity that includes β-lactamase enzymes of classes A, C, and selected D examples. In this work acylation and deacylation rates were measured against the clinically important enzymes CTX-M-15, KPC-2, Enterobacter cloacae AmpC, Pseudomonas aeruginosa AmpC, OXA-10, and OXA-48. The efficiency of acylation (k2/Ki) varied across the enzyme spectrum, from 1.1 × 10(1) m(-1)s(-1) for OXA-10 to 1.0 × 10(5) for CTX-M-15. Inhibition of OXA-10 was shown to follow the covalent reversible mechanism, and the acylated OXA-10 displayed the longest residence time for deacylation, with a half-life of greater than 5 days. Across multiple enzymes, acyl enzyme stability was assessed by mass spectrometry. These inhibited enzyme forms were stable to rearrangement or hydrolysis, with the exception of KPC-2. KPC-2 displayed a slow hydrolytic route that involved fragmentation of the acyl-avibactam complex. The identity of released degradation products was investigated, and a possible mechanism for the slow deacylation from KPC-2 is proposed.
authors.affiliation: From the Infection Innovative Medicines Unit and the Discovery Sciences Innovative Medicines Unit, AstraZeneca R&D Boston, Waltham, Massachusetts 02451.
authors.forename: David E
authors.initials: DE
authors.lastname: Ehmann
authors.forename: Haris
authors.initials: H
authors.lastname: Jahic
authors.forename: Philip L
authors.initials: PL
authors.lastname: Ross
authors.forename: Rong-Fang
authors.initials: RF
authors.lastname: Gu
authors.forename: Jun
authors.initials: J
authors.lastname: Hu
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Durand-Réville
authors.forename: Sushmita
authors.initials: S
authors.lastname: Lahiri
authors.forename: Jason
authors.initials: J
authors.lastname: Thresher
authors.forename: Stephania
authors.initials: S
authors.lastname: Livchak
authors.forename: Ning
authors.initials: N
authors.lastname: Gao
authors.forename: Tiffany
authors.initials: T
authors.lastname: Palmer
authors.forename: Grant K
authors.initials: GK
authors.lastname: Walkup
authors.forename: Stewart L
authors.initials: SL
authors.lastname: Fisher
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry; D053961:Azabicyclo Compounds / Q000737:chemistry*; D024881:Drug Resistance, Bacterial; D016972:Enterobacter cloacae / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry; D004926:Escherichia coli / Q000187:drug effects* / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008826:Microbial Sensitivity Tests; D010957:Plasmids / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000378:metabolism; D013997:Time Factors; D001618:beta-Lactamases / Q000737:chemistry*
chemical_list: D000900:Anti-Bacterial Agents; D053961:Azabicyclo Compounds; D004791:Enzyme Inhibitors; C543519:avibactam; C473109:beta-lactamase CTX-M-15; C491478:beta-lactamase KPC-2; C055187:beta-lactamase PSE-2; D001618:beta-Lactamases; C108449:oxacillinase
keywords: Antibiotics; Drug Discovery; Enzyme Inhibitors; Enzyme Kinetics; Mass Spectrometry (MS)
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