Multi-target benzylidene camphor derivatives: selective cytotoxicity, acetylcholinesterase inhibition, and via a ROS-associated mitochondrial effects in AGS cells.

This study describes the synthesis and biological evaluation of ten benzylidene camphor derivatives as potential dual-target agents with cytotoxic and acetylcholinesterase (AChE) inhibitory activities. The compounds were synthesized via microwave-assisted Claisen-Schmidt condensation with high yields (70-95%) and fully characterized by spectroscopic techniques. Cytotoxic activity was evaluated against human tumor (AGS, HT-29, and MCF-7) and non-tumoral cell lines (GES-1 and MCF-10A). Methoxy-substituted derivatives (3-6) emerged as the most effective, showing selective cytotoxicity. Specifically, compound 4 exhibited the highest potency against AGS cells (IC50 = 43.79 ± 0.4 μM) and a selectivity index (SI) of approximately 2.0. Flow cytometry analysis in AGS cells suggests the involvement of a ROS-mediated mitochondrial apoptotic pathway, as evidenced by increased oxidative stress, loss of mitochondrial membrane permeability (ΔΨm), and, for compound 4, significant caspase activation. Furthermore, compounds 5 and 6 showed the strongest AChE inhibition, with IC50 values of 27.3 ± 0.4 μM and 21.0 ± 0.2 μM, respectively. Molecular docking indicated that these derivatives establish stabilizing π-π and hydrophobic interactions within the catalytic gorge. In silico ADME analysis suggested favorable drug-like properties and blood-brain barrier permeability. These findings highlight benzylidene camphor derivatives as promising multifunctional scaffolds for the potential development of agents at the interface of oncology and neurodegeneration.