Integrated Computational Evaluation of 4-Hydroxychalcone as a Potential Acetylcholinesterase Inhibitor

Background Alzheimer’s disease (AD) is a major neurodegenerative disorder characterized by cognitive decline, amyloid-β aggregation, neurofibrillary tangles, and cholinergic dysfunction. Acetylcholinesterase (AChE) inhibition remains an important therapeutic strategy for AD management. Methods The structural, electronic, and anti-Alzheimer potential of 4-hydroxychalcone (4HC) were investigated using experimental and computational approaches. Structural characterization was performed using X-ray diffraction, FT-IR, FT-Raman, and UV–Visible spectroscopy, supported by DFT calculations at the B3LYP/6-311++G(d,p) level. Molecular docking against AChE was followed by 100 ns molecular dynamics simulations, ADMET prediction, and in vitro AChE inhibition assays. Results The HOMO–LUMO energy gap was calculated as 3.89 eV in the gas phase and 3.71 eV in DMSO. Molecular docking revealed strong binding affinity toward AChE with a maximum binding energy of −9.7 kcal/mol. Molecular dynamics simulations confirmed stable ligand–protein interactions. ADMET predictions suggested favorable drug-likeness and blood–brain barrier permeability. In vitro studies demonstrated dose-dependent AChE inhibitory activity with an IC50 value of 36.86 µg/mL. Discussion The combined experimental and computational findings indicate that the electronic structure and reactive regions of 4HC contribute to effective interaction with AChE and support its potential anti-Alzheimer activity. Limitations The present study is limited to computational and in vitro investigations. Further in vivo studies and toxicity evaluations are required to validate the therapeutic efficacy and safety of 4HC. Conclusion 4-Hydroxychalcone exhibits promising AChE inhibitory activity with favourable pharmacokinetic properties, highlighting its potential as a lead scaffold for further development in Alzheimer’s disease therapeutics.