Cytotoxic, Antioxidant, Anti‐Inflammatory, and Apoptotic Activities of Paraben Derivative Tetracyclic Spermine Cyclotriphosphazene Compounds in Human Lung Cancer (A549 Cell)

ABSTRACT Lung cancer is one of the most common and deadliest malignancies worldwide. Multidisciplinary research efforts, ranging from in vitro studies to clinical trials, are being conducted to elucidate its molecular mechanisms and to develop novel therapeutic strategies. In this study, hybrid tetracyclic derivatives integrating paraben, spermine, and cyclotriphosphazene pharmacophores were synthesized. These compounds stand out due to their ability to exert pro‐apoptotic effects, facilitate DNA binding and cellular uptake, and provide chemical stability with versatile substitution potential. Such synergistic features make this scaffold series more promising compared to previously reported derivatives. The cytotoxic activities of six newly synthesized molecules ( 1–6 ) were evaluated using the MTT assay in A549 human lung cancer and MRC‐5 normal lung fibroblast cells. In A549 cells, IC 50 values ranged from 12.210 to 35.150 µM, with compound 5 (IC 50 : 12.650 µM, LogIC 50 : 1.102) showing the strongest cytotoxic effect, while minimal toxicity was observed in MRC‐5 cells. qRT‐PCR analysis revealed that compound 5 significantly upregulated CAT (1.52‐fold) and SOD (6.63‐fold) expression, suppressed IL‐1β, and reduced BCL‐2 levels. Compound 3 also enhanced SOD (3.10‐fold) and decreased NFκB (0.7‐fold) expression. Enzyme‐linked immunosorbent assay (ELISA) results confirmed these findings: compounds 5 and 6 exhibited the strongest antioxidant activity by reducing GSH levels to 57.79% and 57.68% of control, respectively, while compound 3 exerted the greatest anti‐inflammatory effect (IL‐1β: 61.56% of control). Importantly, compound 5 demonstrated the highest apoptotic activity with a 139.20% increase in Caspase‐3 protein levels. Among them, compound 5 demonstrated the strongest overall efficacy, markedly reducing cell viability while modulating oxidative stress and apoptotic signaling pathways. These findings highlight the potential of this newly designed scaffold as a promising platform for future anticancer drug development.