Integrating Network Pharmacology and Optimization of Preparation Methods to Enhance the Anticancer Effect of Cordyceps militaris on Lung Cancer

Background/Aim: Cordyceps militaris, a medicinal fungus, is renowned for its anticancer properties. To maximize its therapeutic efficacy, this study aimed to optimize the extraction and preparation methods of Cordyceps militaris and evaluate its effects on non‑small cell lung cancer (NSCLC). Additionally, the study explored the potential of natural products as novel therapeutic agents. Materials and Methods: Network pharmacology was employed to identify key compounds in Cordyceps militaris, including cordycepin and adenosine. Various extraction conditions, including temperature (50 to 121) and duration (up to 72 h), were systematically tested. Freeze‑drying and spray‑drying methods were compared for their ability to preserve the bioactivity of the extracts. Lung cancer cell lines (LLC1, H460, H1299) were treated with the optimized extracts, and their effects on cell viability, migration, and colony formation were evaluated. Apoptotic markers were analyzed using western blotting. Results: Cordyceps militaris extracts optimized at 80 for 24 h demonstrated the highest levels of cordycepin (505.59 ppm) and adenosine (81.80 ppm). Freeze‑dried extracts obtained using this method exhibited significantly greater anticancer activity compared to spray‑dried extracts. The optimized extract suppressed proliferation, migration, and clonogenic potential of lung cancer cells in a dose‑dependent manner. Apoptosis‑related pathways were activated, as evidenced by enhanced caspase‑3 and PARP cleavage. Network pharmacology and molecular docking confirmed that cordycepin and adenosine target key proteins, including P53 and EGFR, to induce apoptosis. Conclusion: This study underscores the importance of optimizing extraction and drying methods to enhance the therapeutic potential of Cordyceps militaris. Furthermore, the findings highlight its promise as a natural anticancer agent, particularly for lung cancer.