Novel benzofuran/pterostilbene hybrids trigger programmed cell death and impair migration in CRC cells

Colorectal cancer (CRC) remains one of the most prevalent and lethal malignancies worldwide, highlighting the urgent need for developing effective treatments. Molecular hybridization is a promising strategy for identifying new bioactive compounds. This study focused on designing and synthesizing a novel series of benzofuran-pterostilbene hybrid molecules. These compounds were successfully obtained, and their structures were elucidated by spectroscopic analysis. In addition, the activity of the hybrids was evaluated against colorectal adenocarcinoma cells. After the treatments, hybrids 6d and 6e exhibited the highest activity, with GI 50 values of 11.93 ± 2.38 µM and 4.74 ± 0.38 µM, respectively, suggesting antiproliferative effects and measurable cytotoxicity under the tested conditions. Additionally, Hoechst 33342 fluorescence imaging revealed chromatin condensation and nuclear fragmentation, along with a diffuse DiOC₆ fluorescence pattern relative to the control, suggesting a form of programmed cell death, which was further supported by flow cytometric analysis showing an increased proportion of hypodiploid cells following propidium iodide staining. In parallel, wound-healing assays demonstrated impaired migration and cytotoxic effects that affected cell viability and structural integrity. Molecular docking simulations showed that compounds 6d and 6e bind strongly to mutant p53, CDK4, and PARP-1 proteins, which, in turn, may explain at the molecular level the in vitro cytotoxic effect of these compounds in SW480 colon cancer cells. Lastly, pharmacokinetic and toxicological modelling suggests that hybrids 6d and 6e possess optimal biopharmaceutical profiles with no major safety concerns. All these findings highlight the potential of the benzofuran-pterostilbene scaffold, with compounds 6d and 6e emerging as strong candidates for further evaluation against colorectal cancer.