Artesunate-transferrin(ATS-Tf) adduct was prepared through drug-ligand self-assembly.UV-vis spectrum analysis showed that ATS-Tf adduct can be easily formed with relatively high binding constant at neutral pH(3.4×105 L/mol at pH=7.4).However,the adduct became less stable with low binding constant at acidic condition(1.7×104 L/mol at pH=5.5).Proliferation inhibition studies of ATS-Tf adduct on cancer cells and normal cells showed that the ATS-Tf adduct had better antitumor activity on human to hepatocellular carcinoma cell (HepG2),lung adenocarcinoma cell (A549),and gastric carcinoma cell(MGC-803),while it had low toxicity on normal human liver cell(L-02).HPLC analysis confirmed that the incorporation of Tf increased the uptake of ATS by cancer cells.The interactive model and mechanism of ATS with Tf were further studied by molecular docking and fluorescence spectroscopy analysis.The possible inhibition mechanism of ATS-Tf adduct on cancer cells was also proposed.