title: Penicillin sulfone inhibitors of class D beta-lactamases.
abstract: OXA beta-lactamases are largely responsible for beta-lactam resistance in Acinetobacter spp. and Pseudomonas aeruginosa, two of the most difficult-to-treat nosocomial pathogens. In general, the beta-lactamase inhibitors used in clinical practice (clavulanic acid, sulbactam, and tazobactam) demonstrate poor activity against class D beta-lactamases. To overcome this challenge, we explored the abilities of beta-lactamase inhibitors of the C-2- and C-3-substituted penicillin and cephalosporin sulfone families against OXA-1, extended-spectrum (OXA-10, OXA-14, and OXA-17), and carbapenemase-type (OXA-24/40) class D beta-lactamases. Three C-2-substituted penicillin sulfone compounds (JDB/LN-1-255, JDB/LN-III-26, and JDB/ASR-II-292) showed low K(i) values for the OXA-1 beta-lactamase (0.70 +/- 0.14 --> 1.60 +/- 0.30 microM) and demonstrated significant K(i) improvements compared to the C-3-substituted cephalosporin sulfone (JDB/DVR-II-214), tazobactam, and clavulanic acid. The C-2-substituted penicillin sulfones JDB/ASR-II-292 and JDB/LN-1-255 also demonstrated low K(i)s for the OXA-10, -14, -17, and -24/40 beta-lactamases (0.20 +/- 0.04 --> 17 +/- 4 microM). Furthermore, JDB/LN-1-255 displayed stoichiometric inactivation of OXA-1 (the turnover number, i.e., the partitioning of the initial enzyme inhibitor complex between hydrolysis and enzyme inactivation [t(n)] = 0) and t(n)s ranging from 5 to 8 for the other OXA enzymes. Using mass spectroscopy to study the intermediates in the inactivation pathway, we determined that JDB/LN-1-255 inhibited OXA beta-lactamases by forming covalent adducts that do not fragment. On the basis of the substrate and inhibitor kinetics of OXA-1, we constructed a model showing that the C-3 carboxylate of JDB/LN-1-255 interacts with Ser115 and Thr213, the R-2 group at C-2 fits between the space created by the long B9 and B10 beta strands, and stabilizing hydrophobic interactions are formed between the pyridyl ring of JDB/LN-1-255 and Val116 and Leu161. By exploiting conserved structural and mechanistic features, JDB/LN-1-255 is a promising lead compound in the quest for effective inhibitors of OXA-type beta-lactamases.
authors.affiliation: Departments of Pathology, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA.
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Drawz
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Bethel
authors.forename: Venkata R
authors.initials: VR
authors.lastname: Doppalapudi
authors.forename: Anjaneyulu
authors.initials: A
authors.lastname: Sheri
authors.forename: Sundar Ram Reddy
authors.initials: SR
authors.lastname: Pagadala
authors.forename: Andrea M
authors.initials: AM
authors.lastname: Hujer
authors.forename: Marion J
authors.initials: MJ
authors.lastname: Skalweit
authors.forename: Vernon E
authors.initials: VE
authors.lastname: Anderson
authors.forename: Shu G
authors.initials: SG
authors.lastname: Chen
authors.forename: John D
authors.initials: JD
authors.lastname: Buynak
authors.forename: Robert A
authors.initials: RA
authors.lastname: Bonomo
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology; D020134:Catalytic Domain; D002509:Cephaloridine / Q000737:chemistry; D002511:Cephalosporins / Q000737:chemistry / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D007700:Kinetics; D008826:Microbial Sensitivity Tests; D008958:Models, Molecular; D010068:Oxacillin / Q000737:chemistry; D010406:Penicillins / Q000737:chemistry / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors; D013379:Substrate Specificity; D013450:Sulfones / Q000737:chemistry / Q000494:pharmacology; D018440:beta-Lactam Resistance; D065093:beta-Lactamase Inhibitors; D001618:beta-Lactamases / Q000737:chemistry / Q000145:classification
chemical_list: D000900:Anti-Bacterial Agents; D002511:Cephalosporins; D004791:Enzyme Inhibitors; D010406:Penicillins; D011994:Recombinant Proteins; D013450:Sulfones; D065093:beta-Lactamase Inhibitors; C039404:beta-lactamase OXA-2; D001618:beta-Lactamases; D002509:Cephaloridine; D010068:Oxacillin