Weifuchun (WFC) is a kind of Chinese patent medicine, which is mainly used to treat chronic gastritis, gastric ulcer and other gastric diseases. However, the mechanism of Weifuchun in the treatment of Colorectal cancer (CRC) remains unclear.To investigate Weifuchun's anti-cancer efficacy against colorectal carcinoma and explore its underlying molecular mechanisms.The TCMSP and BATMAN-TCM databases were employed to filter WFC's bioactive ingredients, while DrugBank provided information on their respective protein targets. The GeneCards and OMIM databases were queried to obtain CRC-associated targets, which were then cross-referenced with WFC targets via Venn diagram analysis to identify common targets. A network pharmacology approach was employed, using Cytoscape to visualize TCM compound-target-disease relationships, with subsequent functional and signaling pathway evaluation. Cell proliferations were detected by CCK-8, LDH, and colony forming assays. Flow cytometric analysis and Hoechst 33258 were used to detect apoptosis. Transwell assays were employed to detect the cell migration. AKT signaling pathway related protein expressions were detected by Western blot. Finally, immunohistochemistry and Western blot were analyzed to evaluate Ki-67 expression and AKT signaling pathway related proteins in the xenograft mouse model. This retrospective cohort study enrolled colorectal cancer patients who underwent curative-intent surgery. Based on whether they received treatment with WFC after surgery, the patients are divided into two groups: the WFC combined chemotherapy group (WFC group) and the chemotherapy-only group (control group). Propensity score matching (PSM) was employed to balance baseline characteristics between the two groups, enabling comparison of recurrence/metastasis rates.Our analysis identified 67 bioactive compounds, 211 therapeutic protein targets, 1699 disease-associated targets, with 97 overlapping targets and 187 relevant signaling pathways. Bioinformatics analysis through GO and KEGG databases revealed WFC's regulatory effects on cellular processes including cell migration, adhesion, apoptosis and cell cycle. Among the top 20 tumor-associated pathways identified, the targets predominantly localized to the PI3K/Akt and NF-κB signaling pathways. In vitro, WFC could significantly inhibit cell proliferation, migration, and induce apoptosis in HCT116 and SW620 cells. Molecular mechanism showed decreases in AKT, p-AKT, p65, p-65 and p-IκB proteins, and an increase in IκB protein level were observed in WFC treated-HCT116 and SW620 cells. Furthermore, WFC demonstrated significant inhibition of colorectal cancer (CRC) tumor growth in a xenograft mouse model, concomitant with downregulation of the AKT signaling pathway. Pilot exploratory analysis observed a trend toward lower recurrence and metastasis rates at 1, 3, and 5 years among patients receiving the combination therapy.WFC may exert its anti-tumor effect through modulation of the AKT and NF-κB signaling pathways, and then regulate proliferation, migration and apoptosis in colorectal cancer cells.Copyright © 2025. Published by Elsevier B.V.