Objective Study the anti-tumor effects in vivo of AMH-D on S-180 cell lines, the synergistic effects of AMH-D and cyclophosphamide,and investigate the way and its strength of the effect. Provide the basis for the development of anti-cancer drug. Methods Kunming mice were transplanted with S-180 tumor cells subcutaneously in the right armpit. Intraperitoneal injection was done after randomization on the next day. Mice were killed on the eleventh day, and tumors were stripped and weighed. The tumor weight was used as indicator for analysis and evaluation. Results The results showed that AMH-D could effectively inhibit the growth of S180 cells transplanted tumor. The tumor inhibition rate was 50.45%at the dose of 150 mg/kg, with a dose-effect relationship. There were no obvious impacts on the growth of the weight of mice. The results showed that AMH-D had a synergistic effect combined with cyclophosphamide within a certain dose. Conclusion Fungus extract AMH-D has a great effect on anti-tumor in vivo of S180 cells transplanted tumor, and has a synergistic effect combined with cyclophosphamide within a certain dose.