Ginseng possesses antitumor effects, and ginsenosides are considered to be one of its mainactive chemical components. Ginsenosides can further be hydrolyzed to generate secondary saponins,and 20(R)-panaxotriol is an important sapogenin of ginsenosides. We aimed to synthesize a new gin sengenin derivative from 20(R)-panaxotriol and investigate its antitumor activity in vivo and in vitro. Methods: Here, 20(R)-panaxotriol was selected as a precursor and was modified into its derivatives. Thenew products were characterized by 1H-NMR, 13C-NMR and HR-MS and evaluated by molecular docking,MTT, luciferase reporter assay, western blotting, immunofluorescent staining, colony formation assay,EdU labeling and immunofluorescence, apoptosis assay, cells migration assay, transwell assay and in vivoantitumor activity assay. Results: The derivative with the best antitumor activity was identified as 6,12-dihydroxy-4,4,8,10,14-pentamethyl-17-(2,6,6-trimethyltetrahydro-2H-pyran-2-yl)hexadecahydro-1H-cyclopenta[a]phe nanthren-3-yl(tert-butoxycarbonyl)glycinate (A11). The focus of this research was on the antitumor ac tivity of the derivatives. The efficacy of the derivative A11 (IC50 < 0.3 mM) was more than 100 timeshigher than that of 20(R)- panaxotriol (IC50 > 30 mM). In addition, A11 inhibited the protein expressionand nuclear accumulation of the hypoxia-inducible factor HIF-1a in HeLa cells under hypoxic conditionsin a dose-dependent manner. Moreover, A11 dose-dependently inhibited the proliferation, migration,and invasion of HeLa cells, while promoting their apoptosis. Notably, the inhibition by A11 was moresignificant than that by 20(R)-panaxotriol (p < 0.01) in vivo. Conclusion: To our knowledge, this is the first study to report the production of derivative A11 from20(R)-panaxotriol and its superior antitumor activity compared to its precursor. Moreover, derivative A11can be used to further study and develop novel antitumor drugs.