Abstract: OBJECTIVE: Synthesis and characterization of disulfiram (DSF) derivatives,and preliminary study on their antitumor activities in vitro. METHODS: Two derivatives were synthesized in two steps,and their structures were confirmed using ¹H NMR,¹³C NMR and LC-MS. CCK-8 cytotoxicity assay was used to compare the inhibitory effects of DSF and its derivatives on proliferation of MCF-7 and A549 cells with or without FBS. Effects of NAC on anti-tumor activities of these compounds were detected in the same way. Whether DSF and its derivatives could induce apoptosis were analyzed using flow cytometry. Expressions of related apoptotic proteins in MCF-7 and A549 cells were detected using Western blot. RESULTS: Results from the spectral analyses show that the structures of the derivatives were consistent with the expectation. The CCK-8 results show that the inhibitory effects of the two derivatives on these two kinds of tumor cells were stronger than that of DSF,and there were significant differences on the value of IC₅₀. There was an obvious decline in the proliferation inhibitory effect of the derivatives in the absence of FBS. In addition,the inhibitory effects on the proliferation of MCF-7 and A549 cells was significantly weakened after the addition of NAC (P < 0.05). Flow cytometry analyses show that the apoptosis rates of MCF-7 and A549 cells which were treated with the two derivatives increased compared with DSF (P < 0.05 or P < 0.01). The results from Western blot analyses show that the two derivatives increased the expression of BAX and decreased the expression of PARP in the MCF-7 and A549 cells. CONCLUSION: The two derivatives showed anti-tumor activities. In addition,the inhibitory effects on the two kinds of tumor cells in the presence of FBS were better than that of DSF. The mechanisms may be related to the expression of apoptosis-related proteins and the increase of intracellular ROS production.

Key words: disulfiram, derivatives, antitumor, apoptosis related proteins