<p>Non-small cell lung cancer (NSCLC) accounts for approximately 85% of all lung cancer cases and has gained considerable global attention. Epidermal growth factor receptor (EGFR) plays a key role in NSCLC treatment. NSCLC with wild-type EGFR (WT-EGFR) is not responsive to EGFR tyrosine kinase inhibitors (TKIs), and chemotherapy, such as cisplatin (cDDP), continues to be utilized as a primary clinical treatment. However, the therapeutic efficacy of cDDP remains limited. Therefore, there exists an urgent need for the development of novel therapeutic strategies. Cudratricusxanthone A (CTXA) as a natural bioactive xanthone exhibits potential antitumor activity, but the mechanism has been unknown. In this study, we aimed to evaluate the potential antitumor effect and mechanism of CTXA against NSCLC with WT-EGFR by cell experiments, molecular dynamics simulation and surface plasmon resonance (SPR) technology. Our results showed that CTXA inhibited cell proliferation in NSCLC cells by suppressing the EGFR/Erk/AKT pathway. Additionally, CTXA exhibited antitumor effects by inhibiting the migration of A549 cells, causing G1 phase arrest, and inducing cell apoptosis. Further investigation revealed that these effects were mediated by the binding of CTXA to EGFR, with a K_D value of 6.302 × 10^− 5 M. This interaction may lead to the suppression of EGFR phosphorylation and its downstream signaling. Moreover, the synergistic inhibition of CTXA in combination with cDDP was also found to be mediated by the suppression of the EGFR pathway and its downstream signaling. We demonstrated that CTXA exhibited antitumor activity and enhanced chemosensitivity to cDDP against NSCLC via the EGFR signaling. Our findings indicate that the utilization of CTXA alone or in combination with cDDP represents a potential novel candidate against NSCLC harboring WT-EGFR.</p>