Rational design, synthesis, biological evaluation, molecular docking, and molecular dynamics of substituted uracil derivatives as potent anti-cancer agents.

[Display omitted] • Novel uracil derivatives synthesized as potential anticancer agents. • Compounds 5, 11, and 15 showed potent activity against HepG-2, HCT-116, and MCF-7 cell lines. • Compound 15 induced apoptosis by modulating Bax, BCL-2, PI3K, and STAT1 proteins. • Molecular docking and dynamics confirmed stable binding to thymidylate synthase. • Uracil derivative 15 is a promising candidate for cancer therapy development. An efficient synthesis of a series of uracil analogous was performed to obtain new potential anticancer agents. The cytotoxic effect of the synthesized derivatives was assessed in vitro against three cancer cell lines, namely hepatic cancer (HepG-2), colon cancer (HCT-116), and breast cancer (MCF-7). Among the tested compounds, 5 , 11 and 15 stood as potent uracil derivatives with pan cytotoxicity against the 3 cell lines out-performing the reference compound 5-FU. Furthermore, selected compounds underwent thymidylate synthase (TS) enzyme inhibition assay and demonstrated effective inhibition of the enzyme's catalytic activity. Thereafter, flow cytometric apoptosis and protein expression of pro- and anti-apoptotic markers Bax, BCL-2, PI3K, and STAT1 proteins assays were performed employing the most active compound on the respective most responsive cell line and compounds demonstrated effectiveness in inducing apoptosis in the treated cell lines. Finally, in silico studies encompassing molecular docking and molecular dynamics studies were also conducted to predict the interaction mechanisms and stability of the active compounds within the active site of their biological target TS. [ABSTRACT FROM AUTHOR]