This study aims to investigate the chemical constituents of the Uyghur medicinal herb Hyssopus cuspidatus Boriss. Isolation and purification were carried out using column chromatography techniques such as silica gel, Sephadex LH-20, and ODS, combined with semi-preparative HPLC. The structures of the isolated compounds were elucidated based on extensive spectroscopic analysis. In addition, the anti-inflammatory activities of the obtained compounds were evaluated using the MTT assay.  Eighteen compounds were isolated from the 95% ethanol extract of H. cuspidatus and identified as: (1S, 2R, 5S)-2-hydroxy-2-(hydroxymethyl)-6, 6-dimethylbicyclo [3, 1, 1] heptan-3-one (1), 19, 20-epoxy-12-methoxy-11, 14, 19-trihydroxy-7-oxo-8, 11, 13-abietatriene (2), (1R, 1′R, 5S, 5′S)-6, 6, 6′, 6′-tetramethyl [2, 2′-bicyclo [3, 1, 1] heptane]-2, 2′-diol (3), methyl 4-hydroxy-6, 6-dimethylbicyclo [3, 1, 1] hept-2-ene-2-carboxylate (4), 4-epianol (5), β-sitosterol (6), ursolic acid (7), 2, 3-dihydro-6, 7-dimethoxy-2-methyl-1H-inden-1-one (8), (2R)-5-hydroxy-7, 2', 3'-trimethoxyflavone (9), 4'-hydroxy-5, 6, 7-trimethoxyflavone (10), bergapten (11), hydroxydihydrobovolide (12), pinic acid (13), Ethyl (E)-3-(4-hydroxyphenyl)acrylate (14), ferulic acid (15), 1-feruloylglycerol (16), syringyl alcohol (17), salicylic acid (18). Compounds 3-5, 8, 9, 12, 14 and 17 are isolated from H. cuspidatus for the first time. Compounds 2-4 and 9 exhibited significant anti-inflammatory activity, with IC₅₀ values of 0.15-0.39 μg/mL against nitric oxide release in lipopolysaccharide-induced RAW 264.7 cells.