The ADvANCE project, funded by the German Federal Ministry of Education and Research (BMBF) under grant number 16GW0217, ran from 1 April 2019 to 31 March 2023 and aimed to develop a new class of natural‑product‑derived inhibitors that block carbapenemase enzymes responsible for antibiotic resistance in Gram‑negative bacteria. The consortium comprised the Helmholtz Zentrum München (HZI), the Helmholtz Institute for Molecular Systems Biology (HMGU), and the Technical University of Munich (TUM). HZI carried out the synthesis of fragment derivatives, while HMGU performed structural investigations, including X‑ray crystallography, nuclear magnetic resonance (NMR) interaction assays, and quantum‑mechanical binding studies. TUM was responsible for project management, the design of biochemically and biologically relevant assays, and the coordination of external synthesis and ADME‑toxicity testing, which was largely outsourced to the contract research organization Enamine.

The scientific effort focused on a fragment library that exhibited inhibitory activity against both serine‑β‑lactamases (SBL) and metallo‑β‑lactamases (MBL). Initial fragments showed modest binding in biolayer interferometry (BLI) assays and limited antibacterial activity. To overcome the high ligand mobility that obscured crystal structures, the team refined the scaffold to improve physicochemical properties, enabling successful co‑crystallization with the IMP‑13 enzyme. Five high‑resolution crystal structures of these optimized derivatives were solved, revealing a deeply buried carboxylate group and a halogen bond that appear to be key for binding. Complementary ^1H‑^15N heteronuclear single quantum coherence (HSQC) experiments with isotopically labeled MBL and SBL confirmed the interaction sites and provided further validation of the binding mode.

An artificial intelligence tool developed at HMGU, trained on over 20 000 protein‑ligand complexes, was employed to propose chemical modifications. The resulting compounds displayed improved physicochemical profiles and retained broad inhibitory activity across carbapenemase classes, although none achieved a dramatic increase in potency against all tested enzymes. Nevertheless, the optimized fragments showed enhanced antibacterial activity in orthogonal growth assays against Escherichia coli and Klebsiella pneumoniae, and the data were used to secure additional funding through the m4 Award and GO‑Bio initial programmes. These subsequent projects further improved activity against Acinetobacter baumannii and Pseudomonas aeruginosa.

The project’s budget totaled approximately €373 000, covering personnel (€162 800), materials (€30 000), external services (€153 900), travel (€900), and administrative costs (€25 801). The ADvANCE effort not only generated a set of structurally characterized inhibitors with promising broad‑spectrum activity but also established a platform for rational design that integrates crystallographic data, quantum‑mechanical insights, and AI‑driven optimization. The collaboration between academic laboratories and a CRO, combined with the support of the BMBF and subsequent funding bodies, positioned the consortium to continue advancing these novel inhibitors toward clinical development.