Exploring the Breast Cancer Inhibitor from Flavone Subclass: In Silico and In Vitro Assays

Abstract

Flavones, a subclass of flavonoids, are widely recognized for their diverse biological activities, including antioxidant, anti-inflammatory, and anticancer effects. Amentoflavone, a naturally occurring biflavonoid, has attracted attention for its potential to modulate cancer-related molecular targets. This study investigated the anticancer potential of amentoflavone against estrogen receptor alpha (ERα), a key therapeutic target in ER-positive breast cancer, using an integrated in silico and cytotoxicity approach. Molecular docking analysis using AutoDock4 demonstrated that amentoflavone binds strongly to ERα with a binding affinity of −37.03 kJ/mol and exhibits interaction similarity with the native ligand 4-hydroxytamoxifen (−48.27 kJ/mol). A 100-ns molecular dynamics simulation confirmed the stability of the amentoflavone–ERα complex, as indicated by stable RMSD, RMSF, SASA, and radius of gyration values, along with favorable MM/PBSA binding energy. Cytotoxicity evaluation using the MTT assay in T47D breast cancer cells showed that amentoflavone exhibited moderate cytotoxicity (IC50 = 169.547 ± 34.1651 µg/mL), supported by dose-dependent apoptotic morphological changes. These findings suggest that amentoflavone has potential as a natural lead compound for further optimization and development of ERα-targeted therapies in ER-positive breast cancer.