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Celastrus rugosus belongs to the family Celastraceae, genus Celastrus. It is a vine-like shrub, widely distributed in Hubei, Guizhou, Sichuan, Yunnan province and Eastern Tibet of China growing at 1400-3600 m above sea level. The roots of C. rugosus have been used as Chinese folk medicine for rheumatism, strain and measles in children. However, up to now, only six flavone glycosides and a cafferic acid have been reported from this plant. In this study, we investigated the chemical constituents of C. rugosus, which collected from Xishuangbanna, Yunnan province. Thirty-five compounds were isolated by various chromatographic methods (silica gel column chromatography, preparative-TLC, Sephadex LH-20 column chromatography, preparative-HPLC, et al.), the structures of all compounds were identified as follows by the spectroscopic analysis (ESI-MS, HRESI-MS, IR, 1H-NMR, 13C-NMR, DEPT, 2D-NMR, and et al) : *CR-1: Rugosusine A *CR-2: Rugosusine B CR-3: Euonymine CR-4: Wilfornine G CR-5: Putterine B CR-6: 4-Hydroxy-7-epi-chuchuhuanine E-V CR-7: Paniculatine A CR-8: Euonine CR-9: 20-epi-katonic acid CR-10: 3β, 22 -dihydroxy-olean-12-en-29 oic acid CR-11: 2 , 3β-Dihydroxy-lup-20(29)-ene CR-12: Inulacappolide CR-13: 3, 4′, 5-Trihydroxy-2′, 5′, 6, 7-tetramethoxyflavone CR-14: Brickellin CR-15: 5-Hydroxy-3′, 4′, 6, 7-tetramethoxy flavonol CR-16: Epiafzelechin CR-17: (?)-Epicatechin CR-18: (+)-Catechin CR-19: (3R)-Mellein CR-20: N-(N-Benzoyl-L-phenylalanyl-)-O-actyl-L-phenylalanol CR-21: N(N′-Benzoyl-S*-phenylalaninyl)-S*-phenylalaninol benzoate CR-22: Rumejaposide CR-23: Cassialoin CR-24: Dibutyl phthalate CR-25: Bis (2-ethylhexyl) phthalate CR-26: Diisobutyl phthalate CR-27: 3, 5-Dimethoxy-4-hydroxy-benzaldehyde CR-28: Vanillin CR-29: 5-(Methoxymethyl) resorcinol CR-30: 4-Hydroxy-3, 5-dimethoxybenzenemethanol CR-31: 2, 4, 6-trimethoxy-1-O-β-D-glycoside CR-32: 2, 5-Bis(hydroxyethyl)-1,4-bismethoxybenzene CR-33: N-(2-Aminophenyl) urea CR-34:β-sitosterol CR-35: DaucosterolAmong them, CR-1 and CR-2 were two newβ-dihydroagarofuran alkaloids. Others were isolated from this plant for the first time.Furthermore, 26 compounds isolated from this plant were evaluated for their cytotoxicity against two human cancer cell lines (SKOV-3 and MGC-803) and one African green monkey kidney cell lines (COS-7) using CCK-8 method. All compounds showed weak cytotoxic activity against SKOV-3 and MGC-803 cell line and no inhibitory activity was observed with COS-7 cell lines.
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