This study explores the anti-inflammatory potential of an endophytic fungus, Trametes versicolorCL-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, including11 terpenes, 7flavonoids, 10cinnamic acid derivatives, 6oligopeptides, and 9fatty 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 1within CL-E with the NLRP3 inflammasome (PDB ID: 6NPY). These findings suggest T.versicolorCL-1 as a promising source of natural anti-inflammatory agents and unveil R.roxburghiias a potential reservoir for discovering novel bioactive metabolites.