Antitumor evaluation of novel alizarin-based derivatives through biological and computational approaches.

Two series of alizarine derivatives containing vanillin scaffold (10a-h) or aromatic amide function (12a-h) were synthesized and structurally characterized. The cytotoxic evaluation revealed higher activity towards leukemia cancer cell lines (K562 and HL-60) than solid tumor cells (HeLa and MCF-7). The compound 10 h, containing a benzyl group, showed the most prominent activity against K562 cells, and the lowest toxicity towards healthy cells among all active derivatives. The most active compounds 10f, 10 h, and 12 h were further investigated and induced a significant increase in the percentage of HeLa, K562, and HL-60 cells in the subG1 cell cycle phase in comparison with the control cells. Compounds 10f and 10 h activated apoptosis in K562 cells through all three tested caspases, while derivative 12 h only induced the activation of the main effector caspase-3. Molecular docking simulations suggest that these compounds can form stable complexes with caspase-3, consistent with their experimentally confirmed involvement in caspase-dependent apoptotic pathways. All three tested derivatives demonstrated moderate to strong binding to bovine serum albumin (BSA), with preferential occupation of subdomain IIA (site I), as supported both experimentally and through docking studies. The interaction study of these compounds with DNA indicated their ability to interact with ct-DNA through the minor groove.