To study the anti-inflammatory constituents from the EtOAc extract of Calonyction muricatum leaves based on lipopolysaccharide (LPS)-induced mouse macrophages (RAW 264.7) as the cell model in vitro.Under the guidance of bioactivity screening,the phytochemistry investigation of active EtOAc extract was carried out by silica gel,MCI,ODS,HW-40C column chromatography and semi-prep HPLC methods.The structures of isolated compounds were identified on the basis of physicochemical properties and modern spectroscopy technologies.A total of 15 compounds were obtained from C. muricatum leaves,including 12-oxo-phytodienoic acid（1）,(9Z,12Z,14E,16R)-16-hydroxyoctadeca-9,12,14-trienoic acid（2）,(9Z,12S,13Z,15Z)-12-hydroxyoctadeca-9,13,15-trienoic acid methyl ester（3）,(9Z,11Z,13R,15Z)-13-hydroxyoctadeca-9,11,15-trienoic acid（4）,(9R,10E,12Z)-9-hydroxyoctadeca-10,12-dienoic acid（5）,(9S,10Z,12Z)-9-hydroxyoctadeca-10,12-dienoic acid（6）,1-dodecanol (7),kaempferol-3-O-galactoside (8),6-methoxykaempferol-3-O-galactoside (9),astragaline (10),6-methoxykaempferol-3-O-glucoside (11),(6S,9R)-roseoside (12),ethyl caffeate (13),maltol (14) and hydroxydihydrobovolide (15).Among them,compounds 1-12,14 and 15 were isolated from the genus Calonyction for the first time.Additionally,the bioassay results showed that compounds 1,3,4 and 5 all had certain anti-inflammatory actions.Compounds 1 and 5 significantly inhibited NO production with IC₅₀ values of 1.17 ± 0.51 and 0.97 ± 0.89 μM respectively.