To investigate the chemical constituents and anti-tumor activity of Isodon suzhouensis, a total of 27 compounds were isolated and purified by using various chromatography techniques,including silica gel,ODS,MCI,macroporous adsorption resin and Sephadex LH-20 column chromatography.Structural identification was conducted using modern spectroscopic techniques,including ¹H NMR,¹³C NMR,and MS,their structures were identified as glaucocalyxin D (1),glaucocalyxin B (2),wangzaozin A (3),pseurata A (4),14β-acetyloxy-3α,7α-dihydroxy-ent-kaur-16-en-15-one (5),7α-acetoxy-3α,14β-dihydroxy-ent-kaur-16-en-15-one (6),glaucocalyxin A (7),ursolic acid (8),oleanic acid (9),2α-hydroxyursolic acid (10),3β-hydroxy-oleana-11,13(18)-dien-28-oic acid (11),camaldulenic acid (12),2α,3β-dihydroxy-taraxer-20-en-28-oic acid (13),α/β-amyrin (14,15),11α,12α-oxidotaraxerol (16),Stigmast-4-en-6β-ol-3-one (17),β-sitostenone (18),β-sitosterol (19),3,5,6,7,8,3′,4′-heptamethoxy flavone (20),nobiletin (21),aurantiamide acetate (22),rosmarinic acid (23),blumenol A (24),ethyl caffeate (25),P-hydroxybenzoic acid (26),palmitic acid (27).Except for compounds 2,3,7-10,19,and 27,all other compounds were isolated from Isodon suzhouensis for the first time.The anti-tumor activity of these compounds was assessed using the MTT assay on LNCaP,K562,A549,and Hela.The results indicated that compounds 1,2,3,6 and 26 exhibited significant inhibitory effects on tumor cell growth.