Objective To investigate the chemical constituents of the ethyl acetate fraction from Fissistigma polyanthum. Methods The ethyl acetate fraction of the 95% ethanol extract of F. polyanthum was isolated and purified using silica gel column chromatography, Sephadex LH-20 gel column chromatography, and recrystallization techniques. The structures of the compounds were identified through physicochemical property analysis and nuclear magnetic resonance(NMR) spectroscopic data. Results Twenty-three compounds were isolated from the ethyl acetate fraction of the ethanol extract, identified as 5,6-dihydroxy-7,8-dimethoxyflavone(1), 5,8-dihydroxy-6,7-dimethoxyflavone(2), 6,7,8-trimethoxy-5-hydroxydihydroflavone(3), 5,6,8-trimethoxy-7-hydroxydihydroflavone(4), 5,6-dihydroxy-7-methoxydihydroflavone(5), 5,8-dihydroxy-7-methoxydihydroflavone(6), (-)-epicatechin(7), 2′,3′,5′-trimethoxy-4′,6′-dihydroxychalcone(8), 2′,6′-dihydroxy-3′,4′-dimethoxydihydrochalcone(9), 4′,6′-dihydroxy-2′-methoxydihydrochalcone(10), dihydropedicin(11), 4′,5′-dimethoxy-2′-hydroxy-3′,6′-quinodihydrochalcone(12), aristolactam AII(13), N-trans-feruloyltyramine(14), 4-hydroxy-N-phenethyl-butanamide(15), β-sitosterol(16), daucosterol(17), taraxerol(18), palmitic acid(19), myristic acid(20), 3-phenylpropionic acid(21), 2, 4, 6-trimethoxy-1-O-β-D-glucopyranoside(22), and α-D-glucose(23). Conclusion Compounds 3-6, 8-10, and 15 were isolated for the first time from plants of the genus Fissistigma, while compounds 2-15 were first reported from F. polyanthum.