Rational design of benzobisheterocycle metallo-β-lactamase inhibitors: a tricyclic scaffold enhances potency against target enzymes

Antimicrobial resistance is a global public health threat. Metallo-β-lactamases (MBLs) inactivate β-lactam antibiotics, including carbapenems, are disseminating among Gram-negative bacteria, and lack clinically useful inhibitors. The evolving bisthiazolidine (BTZ) scaffold inhibits all three MBL subclasses (B1 - B3). We report design, synthesis and evaluation of BTZ analogs. Structure-activity relationships identified the BTZ thiol as essential, while the carboxylate is replaceable, with its removal enhancing potency by facilitating hydrophobic interactions within the MBL active site. While the introduction of a flexible aromatic ring is neutral or detrimental for inhibition, a rigid (fused) ring generated nM benzobisheterocycle (BBH) inhibitors that potentiated carbapenems against MBL-producing strains. Crystallography of BBH:MBL complexes identified hydrophobic interactions as the basis of potency towards B1 MBLs. These data underscore BTZs as versatile, potent broad spectrum MBL inhibitors (with activity extending to enzymes refractory to other inhibitors), and provide a rational approach to further improve the tricyclic BBH scaffold.