This study was aimed to analyze the chemical constituents and to screen the antitumor activity of Lindera pulcherrima var. hemsleyana. Seven compounds were isolated from the tuberous roots of L. pulcherrima for the first time, which were identified as Launobine (1), N-Methylhernangerine (2), Nantenine (3), Dehydronantenine (4), (+)-Catechin (5), 8-Hydroxy-9-methoxy-1,2-methylene-dioxyaporphine (6), and Demethylmacrosporine I (7). Their structures were confirmed by spectroscopic data as well as by comparison with those previously reported. These compounds were evaluated for their cytotoxic activity assay toward B16, A549, and HeLa tumor cell lines by the CCK-8 method in vitro. The study of antitumor activity showed that Launobine (1) produced strong inhibitory effect on the growth of melanoma B16 cells(IC50 = 21.06 +/- 0.78 mu M), while compounds 4 and 5 demonstrated weak antitumor activity against melanoma B16 cells.