As a result of anti-inflammatory activity-guided fractionation, 16 secondary metabolites from the underground parts of Valeriana phu L. were obtained, including five new ones belonging to iridoid (1, 2, and 5), phenylpropanoid (6) and neolignan (7) chemical classes. Their structures were elucidated by 1D and 2D NMR analyses as well as HRESIMS. The in vitro anti-inflammatory activities of the extract, fractions and isolates were evaluated through NO inhibition assay on LPS-induced RAW 264.7 cells. Compounds 1-3, 7-9, 11, 13, and 16 which significantly inhibited the nitrite release (IC