Eleven undescribed withanolides, phyminimolides I-XI (1-11), along with eight known derivatives (12-19), were isolated from the whole plant of Physalis minima Linn. Their structures were elucidated by extensive spectroscopic methods, including 1D-NMR, 2D-NMR, HRESIMS, IR, experimental electronic circular dichroism, quantum chemical calculation, and X-ray crystallography. The anti-inflammatory potential of these compounds was assessed in vitro via their capacity to inhibit nuclear factor kappa-B signaling in lipopolysaccharide-stimulated 293T/NF-kappaB-luciferase reporter cells, revealing IC(50) values of 9.95, 3.15, 1.16, 9.73, and 11.74 muM for compounds 4-6, 13, and 15, respectively. Notably, compounds 4 and 13 also demonstrated inhibitory effects on interleukin-6 pathway activation, with IC(50) values of 1.61 and 7.56 muM, further evidencing their anti-inflammatory mechanism. In vivo assessment using dextran sulfate sodium-induced mice model of ulcerative colitis confirmed the translational efficacy of these findings. Compound 15, administered at a dose of 25 mg/kg, significantly ameliorated disease symptoms, as evidenced by a decreased Disease Activity Index score (P < 0.05).