Abstract: OBJECTIVE To explore the potential mechanism and material basis of Dunhuang ancient medical prescription Daxiepi decoction in the treatment of gastric cancer based on chemical bioinformatics method. METHODS Cell proliferation was detected by CCK-8; cell apoptosis and cycle situation was detected by flow cytometry. The Chinese medicine compounds of Daxiepi decoction were downloaded from TCMSP database, Swiss Target Prediction was used to predict the potential targets of compounds. Drugbank, Genecards, TTD and DisGeNET databases were used to obtain gastric cancer targets. Protein interactions were analyzed by STRING database and target clustering was performed using Clusters module. CytoHubba was used for network topology analysis to determine the core targets of Daxiepi decoction in the treatment of gastric cancer, and ClueGO was used for KEGG enrichment analysis of the core targets. Furthermore, molecular docking was used to evaluate the target-compound affinity. dynamic molecular mechanism was study by molecular dynamics simulation. RESULTS The result of cell test showed that the freeze-dried powder of Daxiepi could promote the apoptosis of gastric cancer cells, arrest the cell cycle in the G0-G1 phase(P<0.05), and inhibit its proliferation(P<0.05). A total of 188 targets of Daxiepi decoction and gastric cancer were obtained by network pharmacological analysis. Three types of targets involved in inflammatory response, angiogenesis and cell growth, differentiation were obtained through Clusters module, and mainly involving TNF signaling pathway, PD-L1 expression and PD-1 checkpoint pathway in cancer and p53 signaling pathway and other signaling pathways. Molecular docking results showed that TNF, EGFR and AKT1 had the best affinity with the core components; glyasperinB, licoisoflavanone and 5-prenylbutein showed good affinity with various key potential targets. Through 50 ns molecular dynamics simulations, it was found that the potential active molecules formed stable intermolecular interactions with protein. CONCLUSION This study explores the material basis and molecular mechanism of the potential efficacy of Dunhuang ancient medical prescription Daxiepi decoction on gastric cancer, and provides a chemical bioinformatics reference for the research and clinical application of in treating gastric cancer by “combining cold and heat, and harmonizing yin and yang”.