Eugenol extraction and characterization from clove as a potential anticancer therapeutic agent against breast cancer: Combining an in vitro and in silico molecular docking and dynamics studies

Abstract

Breast cancer (BC) is leading cause of cancer-related mortality among women worldwide, with approximately 2.3 million new cases and 670,000 deaths reported in 2022. Despite advances in therapy, challenges such as drug resistance and adverse side effects necessitate the exploration of alternative treatments, particularly from natural sources. Eugenol (EUG), a bioactive compound derived from clove ( Syzygium aromaticum ), has gained attention for its antioxidant and anticancer properties. In this study, EUG was extracted using ultrasound-assisted extraction and characterized by fourier transform infrared spectroscopy (FTIR), UV-visible spectroscopy, and high-performance thin-layer chromatography. Its antioxidant activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and reducing power assays, while biocompatibility was assessed through a hemolysis assay. Cytotoxicity analysis in MCF-7 BC cells revealed an IC50 of 100 µg/mL, demonstrating dose-dependent inhibition of cell proliferation. The IC50 values for L929 and PBMCs were 3.2 and 2.7mg/mL, respectively, with minimal toxicity toward healthy cells. Mechanistically, EUG induced intracellular reactive oxygen species (ROS) generation, leading to apoptotic cell death. Network pharmacology was performed, and 30 of the most common genes were identified. Molecular docking and molecular dynamics simulation demonstrated strong binding affinities with key proteins. These findings suggest that EUG exhibits significant anticancer potential through ROS-mediated apoptosis and multi-target interactions. Further validation using in vivo animal models, such as xenograft mice, followed by clinical studies, is warranted to establish its therapeutic applicability. GRAPHICAL ABSTRACT