This study explores the anti‐inflammatory potential of an endophytic fungus, Trametes versicolor CL‐1, isolated from the fruit tissues of Rosa roxburghii. Morphological and molecular analyses confirmed the identity of CL‐1. An ethyl acetate extract (CL‐E) from its fermentation broth was subjected to UPLC‐HRMS and GNPS molecular networking. The analysis revealed a diverse array of secondary metabolites, including 11 terpenes, 7 flavonoids, 10 cinnamic acid derivatives, 6 oligopeptides, and 9 fatty acids, as verified by LC‐MS/MS. Notably, CL‐E exhibited significant in vitro anti‐inflammatory activity in RAW264.7 cells. Furthermore, molecular docking studies predicted favorable binding interactions of key compounds 1 within CL‐E with the NLRP3 inflammasome (PDB ID: 6NPY). These findings suggest T. versicolor CL‐1 as a promising source of natural anti‐inflammatory agents and unveil R. roxburghii as a potential reservoir for discovering novel bioactive metabolites.