Synthesis, in-vitro, in-silico, and global DNA methylation studies of curcumin-benzoquinone analog in triple-negative breast cancer (TNBC) cells.

Curcumin is a well-known anticancer agent used for many malignancies; however, its low bioavailability and solubility limit its use in clinical applications. To enhance its efficacy, we synthesized a novel curcumin-benzoquinone analog, JWB1 (3), and evaluated its anticancer potential against triple-negative breast cancer (TNBC) in vitro. We designed JWB1 (3) and structurally identified it using NMR, FTIR, MS, and UV-Vis techniques. The MTT assay was used to evaluate JWB1 (3) cytotoxicity in the MDA-MB-231, MCF-7, and HUVEC cell lines. Flow cytometry was used to examine apoptotic activation and reactive oxygen species (ROS) levels. Global DNA methylation was measured using an ELISA kit. Docking studies and molecular dynamics simulations revealed potential JWB1 (3) interactions with double-stranded DNA (dsDNA). JWB1 (3) showed selective cytotoxicity towards MDA-MB-231 cells (IC(50): 2.94 μg/mL, SI: 23.5 in 24 h), with minimal effects on HUVECs. Treatment with 10 μg/mL JWB1 (3) increased global DNA methylation levels in MDA-MB-231 cells (from 0.87% to 1.92%) more than in MCF-7 cells. The apoptosis assay showed that JWB1 (3) significantly induced MDA-MB-231 cells in the early apoptosis phase (early apoptosis: 45.65% vs. 2.95% in the controls). Furthermore, post-treatment, cancer cells showed a notable decrease in ROS levels. Supported by 100 ns molecular dynamics simulation, molecular docking investigations also showed a stable 3D structure and intercalation of JWB1 (3) with DNA. These findings imply that JWB1 (3) has notable anticancer potential against TNBC by inducing apoptosis, epigenetic modification, and DNA interaction.