Investigation of anticancer and antiinflammatory effects of biologically active artemisinin–benzothiophene derivatives

Cancer is a complex disease that develops as a result of genetic and molecular abnormalities, and the limitations of current treatments, such as toxicity and drug resistance, necessitate the development of new therapeutic agents. In this thesis, the cytotoxic, antiproliferative, anti-migration, anti-invasive, apoptotic, and anti-inflammatory effects of hybrid derivatives obtained by combining artemisinin and benzothiophene scaffolds were investigated on various human cancer cell lines. The study utilized colon (Caco-2), prostate (LNCaP), liver (HepG2), cervical (HeLa), breast (MDA-MB-231, MCF-7), lung (A549), and endometrium (Ishikawa) cancer cells, while healthy HUVEC cells were evaluated as the control group. Cytotoxicity and EC₅₀ values were determined using the MTT assay, proliferation by colony formation, and migration and invasion by scratch-wound and Matrigel invasion assays. Apoptosis was assessed via flow cytometry and RT-PCR, while inflammatory responses were analyzed in RAW264.7 macrophages. Notably, the derivative ÖZK-148, containing a thiophene ring, exhibited low EC₅₀ values in Caco-2 (15.78 μM), HepG2 (17.94 μM), HeLa (36.98 μM), and MDA-MB-231 (31.58 μM) cells. In colony formation assays, ÖZK-144 suppressed growth by more than 80% in Caco-2, LNCaP, and A549 cells. In migration analysis, the derivatives significantly inhibited cell movement into the wound area at 24 and 48 hours. In invasion assays, ÖZK-195 showed the highest inhibition in Caco-2 (87.83%) and HepG2 (89.08%) cells, while ÖZK-170 was most effective in LNCaP (81.23%) cells. Apoptosis assays revealed that the hybrids activated both intrinsic and extrinsic pathways, with increased expression of Bax, Caspase-3, -8, -9, and suppression of Bcl-2. ÖZK-131 demonstrated strong apoptotic activity in LNCaP (80.85%), Ishikawa (82.45%), and HepG2 (65.47%) cells. In anti-inflammatory assays, ÖZK-148, ÖZK-150, and ÖZK-195 reduced NO production and suppressed COX-2, iNOS, IL-6, and TNF-α expression in LPS-stimulated macrophages. In conclusion, artemisinin-benzothiophene hybrids were found to possess strong anticancer potential through their antiproliferative, antimetastatic, apoptotic, and anti-inflammatory properties.