To investigate the chemical constituents of Ferula songarica Pall. ex Schult. and their in vitro antitumor activity, the underground parts were dried, powdered, and extracted with 95% ethanol. Following extraction, the crude material underwent sequential solvent partitioning using petroleum ether, dichloromethane, and ethyl acetate. Subsequent separation and purification of the dichloromethane partition were achieved by combining silica gel column chromatography with semi-preparative high-performance liquid chromatography (HPLC). Thirteen monomeric compounds were ultimately isolated and their structures were elucidated through comprehensive spectroscopic analysis, including nuclear magnetic resonance (NMR) and mass spectrometry (MS), supported by comparison of their physicochemical properties, namely: (1S, 6R, 7R, 8S)-eudesm-5(14)-ene-7-p-hydroxybenzoate (1), dihydrolapidol (2), lancerodiol (3), ferutidin (4), tunetanin A (5), 4-hydroxy-5(E, E-farnesyl)benzoic acid (6), 1-(2, 4-dihydroxyphenyl)-3, 7, 11-trimethyl-3-viny1-6(E), 10-dodecadiene-1-one (7), dshamirone (8), 2,  3-dihydro-7-methoxy-2R, 3R-dimethy1-2-[4, 8-dimethy1-3(E), 7-nonadienyl]-furo[3, 2-c]cou-marin (9), 4, 7-dimethoxy-5-(propanoyl)benzo[d][1, 3]dioxole (10), 4-hydroxy-7-methoxy-5-(propanoyl)benzo[d][1, 3]dio-xole (11), apiol (12), 2-epilaserine (13). Among them, compound 1 was identified as a new eudesmane-type sesquiterpenoid, while compounds 2-13 were isolated from F. songarica for the first time. Additionally, the MTT assay was used to evaluate their in vitro antitumor activity. The results showed that compounds 1, 4, 7 and 13 showed potent inhibitory activity, with IC₅₀ values of 9.17, 13.63, 17.61, 15.52 μmol/L, respectively.