The investigation of the whole plants of Physalis minima led to the isolation of four novel withanolides, designated phyminiolides F-I, together with six known analogues. The structures of 1-4 were elucidated through comprehensive analysis of their spectroscopic data, including HRESIMS, NMR and ECD, while those of 5-10 were identified by comparing their spectroscopic data with the reported literature values. Biologically, compounds 1, 3-4, and 8-10 exhibited moderate anti-inflammatory activities by inhibiting nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated murine RAW 264.7 cells in vitro, with IC50 values ranging from 8.72 to 23.28 μM.