The production of serine ?-lactamase is one of the primary resistance mechanisms used by Gram-negative bacterial pathogens against ?-lactam antibiotics, which include the widely used penicillins and cephalosporins, as well as last resort antibiotics such as the carbapenems. The development of novel ?-lactamase inhibitors is a pressing need underscored by the continuing mutation of ?-lactamases. We propose the development of high affinity non-covalent ?-lactamase inhibitors by targeting conserved structural motifs, particularly those essential for extended spectrum ?-lactamase activity. Prototypes of these inhibitors have already been identified. Specifically, using the CTX-M Class A ?-lactamases as a model system, we aim to: 1) apply a fragment-based and structure-guided approach to develop novel ?-lactamase inhibitor chemotypes;2) study resistance and ligand binding by ultrahigh-resolution and room-temperature X-ray crystallography;and 3) investigate the evolution of resistance mutations against non-covalent inhibitors. These experiments will lead to new ?- lactamase inhibitors with clinical potential, while providing a deeper understanding of ?-lactamase mutations relevant to resistance evolution.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
3R01AI103158-02S1
Application #
8890404
Study Section
Drug Discovery and Mechanisms of Antimicrobial Resistance Study Section (DDR)
Program Officer
Xu, Zuoyu
Project Start
2013-07-01
Project End
2018-06-30
Budget Start
2014-07-17
Budget End
2015-06-30
Support Year
2
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of South Florida
Department
Biochemistry
Type
Schools of Medicine
DUNS #
City
Tampa
State
FL
Country
United States
Zip Code
33612
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Lewandowski, Eric M; Lethbridge, Kathryn G; Sanishvili, Ruslan et al. (2018) Mechanisms of proton relay and product release by Class A ?-lactamase at ultrahigh resolution. FEBS J 285:87-100
Lewandowski, Eric M; Szczupak, ?ukasz; Wong, Stephanie et al. (2017) Antibacterial Properties of Metallocenyl-7-ADCA Derivatives and Structure in Complex with CTX-M ?-Lactamase. Organometallics 36:1673-1676
Pemberton, Orville A; Zhang, Xiujun; Chen, Yu (2017) Molecular Basis of Substrate Recognition and Product Release by the Klebsiella pneumoniae Carbapenemase (KPC-2). J Med Chem 60:3525-3530
Lewandowski, Eric M; Skiba, Joanna; Torelli, Nicholas J et al. (2015) Antibacterial properties and atomic resolution X-ray complex crystal structure of a ruthenocene conjugated ?-lactam antibiotic. Chem Commun (Camb) 51:6186-9
Nichols, Derek A; Hargis, Jacqueline C; Sanishvili, Ruslan et al. (2015) Ligand-Induced Proton Transfer and Low-Barrier Hydrogen Bond Revealed by X-ray Crystallography. J Am Chem Soc 137:8086-95
Nichols, Derek A; Renslo, Adam R; Chen, Yu (2014) Fragment-based inhibitor discovery against ?-lactamase. Future Med Chem 6:413-27