In silico design and development of benzyl piperazine conjugates for targeting choline esterase pathway in Alzheimer's disease

Alzheimer’s disease (AD) is projected to affect 150 million people by 2050 due to global aging. As no cure currently exists and the disease progressively worsens, there is an urgent need for disease-modifying agents capable of restoring cognitive function. The current study aims to identify novel leads for AD treatment using benzyl piperazine hybrids. QSAR analysis was performed using QSARINS to determine structure-activity relationships. Top candidates were selected based on in silico drug-likeness, pharmacokinetics, toxicity profiling, and molecular docking reports. Promising compounds were synthesized, characterized, and evaluated for biological activity. QSAR analysis identified Model 3 as the best-fit model, with a high correlation coefficient (r² = 0.9542) and adjusted r² (r²_adj = 0.9414). Based on predicted activity, 25 new benzyl piperazine derivatives were designed. In silico and docking analysis confirmed strong affinity and drug-like properties for several compounds. Five top hits ( C18, C17, C13, C7 , and C8 ) were selected for synthesis and biological testing. Compound C18 showed potent activity with an IC 50 of 39.10 µg/mL for AChE inhibition and 112.32 µg/mL in the DPPH antioxidant assay, outperforming the reference ascorbic acid (IC 50 = 92.52 µg/mL). All top candidates demonstrated notable neuroprotective effects against glutamate-induced toxicity in SH-SY5Y cells. This integrated computational and experimental approach successfully identified promising AD lead compounds. These findings offer a foundation for further development of effective therapeutics targeting Alzheimer’s disease.