Comparative evaluation of annonacin and 2-Deoxy-D-glucose on lung adenocarcinoma and normal bronchial cells: Differential effects on viability, proliferation and antioxidant defense

Non-small cell lung cancer (NSCLC) exhibits metabolic plasticity that enables it to evade monotherapies targeting a single metabolic pathway. Here, we demonstrate that concurrent inhibition of mitochondrial complex I by annonacin, a plant-derived acetogenin, and glycolysis by 2-Deoxy-D-glucose (2-DG) synergistically compromises NSCLC metabolic resilience and redox homeostasis. A549 (NSCLC) and NL20 (non-malignant) cells were treated with annonacin and/or 2-DG. Cell viability (24 h MTT), clonogenic survival (14 d), superoxide dismutase (SOD), glutathione peroxidase (GPx) activities, and DNA damage (%DNA in tail) using alkaline comet assay were quantified. At 24 h exposure, IC 50 values for annonacin in A549 cells were found to be 6.87 0.11μM and 9.08 0.55 mM for 2-DG; while in NL20 they were 1.944 0.11μM and 1.113 0.013 mM, respectively. Combinations elicited a synergistic response in A549 (Loewe -2.86 to 17.67, mean 8.07) within 2-6 μM annonacin + 2.5-7.5 mM 2-DG combination, and further suppressed clonogenic survival (synergy 10.14 to 22.91, mean 12.39). In both cell lines, SOD and GPx activities were modulated in a dose-dependent manner and were associated with increased DNA damage, indicating that targeting dual metabolic synergistically impaired A549 cell growth and increased genotoxicity. However, between 4-6 μM annonacin + 5-7.5 mM 2-DG, inhibition in NL20 matched or exceeded the values recorded in A549, indicating a dose-limited therapeutic index. Future work integrating in vivo pharmacokinetics, dose optimization, tumor-targeted delivery and diverse cell validation to expand the therapeutic index of the combination treatment and address annonacin’s reported neurotoxicity is desired. • Dual inhibition of glycolysis and OXPHOS synergistically reduced A549 cell viability • Annonacin and 2-DG combination suppressed colony formation in NSCLC cells • Oxidative stress increased via reduced SOD and GPx activities in cancer cells • Normal Bronchial cells showed lower toxicity, indicating selective treatment response • Dual metabolic targeting may overcome resistance in NSCLC through ROS-mediated stress