Repurposed β-lactam compounds as EAAT2 allosteric enhancers: An integrated virtual screening, docking, MD simulation, and ADMET study

Excess synaptic glutamate is neurotoxic, and enhancing its clearance via the excitatory amino acid transporter 2 (EAAT2/GLT-1) represents a promising neuroprotective strategy. This study repurposes a series of β-lactam compounds as potential EAAT2 positive allosteric modulators using an integrated in silico workflow comprising ligand preparation, high-throughput docking (Glide SP/XP), induced-fit docking (IFD), MM-GBSA binding energy estimation, molecular dynamics (MD) simulations, and ADMET prediction (QikProp). Initial virtual screening of 113 analogues identified high-scoring candidates, and subsequent IFD refinement of the top 23 compounds revealed improved complementarity within the EAAT2 allosteric pocket. Six lead analogues were prioritized based on favorable docking scores (−5.36 to −6.83), IFD scores (−755.3 to −759.4), and MM-GBSA binding energies (≈ − 45 to −55 kcal/mol). 100 ns MD simulations confirmed protein–ligand complex stability (protein Cα RMSD ~2 Å; ligand RMSD ~1–3 Å) and persistent interactions, including hydrogen bonding and π–π stacking, mimicking the binding behavior of the known allosteric modulator GT-949. Predicted ADMET profiles indicated drug-likeness, CNS permeability, and low hERG toxicity risk. Overall, this multi-step computational study identifies β-lactam analogues with promising potential as EAAT2 allosteric enhancers, supporting their further experimental evaluation as candidate neuroprotective agents. • Discovery of β-lactam analogues targeting the EAAT2 allosteric site. • Docking and IFD revealed favorable binding of selected compounds. • MD simulations confirmed stable EAAT2–ligand complexes. • Hydrophobic and aromatic interactions drive binding stability. • ADMET profiling suggests promising drug-like properties.