Determination of anticarcinogenic properties of benzofuran endoperoxide derivatives

Cancer, which has begun to emerge as one of the most significant public health issues of our time, is narrowing the gap with cardiovascular diseases each year. This situation has made it crucial to explore alternative approaches alongside existing treatment methods. One such alternative approach is molecular hybridization. Within this framework, various derivatives of benzofuran and artesunat molecules, designed for their anticarcinogenic activity, were synthesized, and their effects on cancer cell lines were investigated. The toxic effects of the synthesized compounds on Caco-2, HepG2, and Ishikawa cell lines were determined using a cell viability test (MTT). The test results showed that only one of the 15 compounds tested had no effect on all cell lines, while the remaining 14 exhibited toxic effects, albeit cell-specific. In particular, BN-80, BN-90, and BN-92, which contain a cyclopentane structure reinforced with an ester bond O molecule, showed a very strong toxic effect on cancer cell lines when compared to the healthy control group. In cell viability tests conducted on the BN-79 molecule, no toxic effects were observed in any cell lines at concentrations below 500 µM (the highest applied concentration). Additionally, molecules with the lowest EC50 values, indicating specific affinity for cell lines, were selected to establish a roadmap for further experiments. In this context, Annexin V staining and qRT-PCR studies showed results below 20% in the Caco-2 cell line, while the substances tested in the Ishikawa and HepG2 cell lines yielded results ranging from 20% to 65%. The effects of the compounds on the cell cycle were investigated using PI staining, and it was determined that the compounds inhibit the transition of cells to the S phase during the cell cycle. Additionally, invasion capacity, wound healing, and colony formation tests were also employed. In addition to the study design, the anti-inflammatory effects of the synthesized compounds were determined using the Griess method and qRT-PCR studies on LPS-stimulated RAW264.7 cell lines. The results of the studies revealed that the synthesized benzofuran-artesunat hybrids possess strong anticancer potential and highlight the significant role these hybrid molecules may play in alternative cancer therapy.