2-mercaptomethyl thiazolidines (MMTZs) inhibit all metallo-β-lactamase classes by maintaining a conserved binding mode

DOI: 10.1021/acsinfecdis.1c00194

Authors: Philip Hinchliffe (University of Bristol) , Diego M. Moreno (Instituto de Química de Rosario (IQUIR, CONICET-UNR); Universidad Nacional de Rosario) , Maria-Agustina Rossi (Instituto de Biología Molecular y Celular de Rosario (IBR, CONICET-UNR)) , Maria F. Mojica (Case Western Reserve University; Louis Stokes Cleveland Department of Veterans Affairs Medical Center; Universidad El Bosque) , Veronica Martinez (Universidad de la República (UdelaR)) , Valentina Villamil (Universidad de la República (UdelaR)) , Brad Spellberg (Los Angeles County and University of Southern California (LAC + USC) Medical Center) , George L. Drusano (University of Florida) , Claudia Banchio (Universidad Nacional de Rosario; Instituto de Biología Molecular y Celular de Rosario (IBR, CONICET-UNR)) , Graciela Mahler (Universidad de la República (UdelaR)) , Robert A. Bonomo (Louis Stokes Cleveland Department of Veterans Affairs Medical Center; Case Western Reserve University School of Medicine; CWRU-Cleveland VAMC Center for Antimicrobial Resistance and Epidemiology (Case VA CARES)) , Alejandro J. Vila (Universidad Nacional de Rosario; Instituto de Biología Molecular y Celular de Rosario (IBR, CONICET-UNR)) , James Spencer (University of Bristol)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Acs Infectious Diseases

State: Published (Approved)
Published: August 2021
Diamond Proposal Number(s): 12342 , 17212

Abstract: Metallo-β-lactamase (MBL) production in Gram-negative bacteria is an important contributor to β-lactam antibiotic resistance. Combining β-lactams with β-lactamase inhibitors (BLIs) is a validated route to overcoming resistance, but MBL inhibitors are not available in the clinic. On the basis of zinc utilization and sequence, MBLs are divided into three subclasses, B1, B2, and B3, whose differing active-site architectures hinder development of BLIs capable of “cross-class” MBL inhibition. We previously described 2-mercaptomethyl thiazolidines (MMTZs) as B1 MBL inhibitors (e.g., NDM-1) and here show that inhibition extends to the clinically relevant B2 (Sfh-I) and B3 (L1) enzymes. MMTZs inhibit purified MBLs in vitro (e.g., Sfh-I, Ki 0.16 μM) and potentiate β-lactam activity against producer strains. X-ray crystallography reveals that inhibition involves direct interaction of the MMTZ thiol with the mono- or dizinc centers of Sfh-I/L1, respectively. This is further enhanced by sulfur-π interactions with a conserved active site tryptophan. Computational studies reveal that the stereochemistry at chiral centers is critical, showing less potent MMTZ stereoisomers (up to 800-fold) as unable to replicate sulfur-π interactions in Sfh-I, largely through steric constraints in a compact active site. Furthermore, in silico replacement of the thiazolidine sulfur with oxygen (forming an oxazolidine) resulted in less favorable aromatic interactions with B2 MBLs, though the effect is less than that previously observed for the subclass B1 enzyme NDM-1. In the B3 enzyme L1, these effects are offset by additional MMTZ interactions with the protein main chain. MMTZs can therefore inhibit all MBL classes by maintaining conserved binding modes through different routes.

Journal Keywords: antibiotic resistance; β-lactamases; inhibitors; carbapenemase

Diamond Keywords: Bacteria; Enzymes

Subject Areas: Biology and Bio-materials, Chemistry, Medicine


Instruments: I03-Macromolecular Crystallography , I04-1-Macromolecular Crystallography (fixed wavelength)

Added On: 09/08/2021 08:58

Documents:
acsinfecdis.1c00194.pdf

Discipline Tags:

Pathogens Antibiotic Resistance Infectious Diseases Health & Wellbeing Biochemistry Chemistry Structural biology Drug Discovery Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)