Caspase-induced Apoptosis as the Major Mechanism underlying the Anticancer Effect of Flavonoids Myricetin and Bergapten: Evidence from an In silico, Network Pharmacology and In vitro Perspective

<title>Abstract</title> <p> Natural phytochemicals have gained increasing attention as potential anticancer agents due to their multi-target activity and favourable safety profiles. The present study aimed to evaluate the anticancer potential of two plant-derived compounds, myricetin and bergapten, using an integrated approach combining <italic>in silico</italic> analyses with <italic>in vitro</italic> validation. The selected compounds were investigated using multiple computational approaches, including bioactivity score (BAS) analysis, pharmacokinetic parameter prediction, SwissTargetPrediction, BOILED-Egg analysis, PASS analysis, principal component analysis (PCA), STRING analysis, KEGG pathway analysis, molecular docking, and molecular dynamics (MD) simulation, along with cytotoxicity assessment using the MTT assay. The results revealed that both compounds exhibited minimal violations of Lipinski’s rule in PASS analysis, where Bergapten displayed zero, and myricetin displayed one violation of Lipinski's parameters. Myricetin showed active bioactivity scores (&gt; 0.0) for kinase inhibitor, nuclear receptor ligand, and enzyme inhibitor, while bergapten exhibited bioactivity scores between − 5.0 and 0.0 for all parameters, indicating moderate biological activity. Drug-likeness evaluation using the OSIRIS Data Warrior v5.2.1 demonstrated favourable pharmacological properties, including good gastrointestinal absorption and permeation. Molecular docking studies indicated strong binding affinities of both compounds toward apoptosis markers, with the highest interaction observed with caspase-6, showing binding energies of -8.36 kcal/mol for myricetin and − 6.33 kcal/mol for bergapten. MD simulation results further confirmed the structural stability of the myricetin-caspase-6 complex. In vitro evaluation demonstrated that both bergapten and myricetin exhibited cytotoxic activity at 40 µM against the A549 lung cancer cell line. Overall, combined computational and experimental findings suggest that myricetin and bergapten may serve as promising lead compounds for anticancer drug development. </p>