To explore potential anti-inflammatory lead compounds, ten new physalin steroids, including three neophysalins (1, 4, and 9) and seven physalins (2, 3, 5-8, and 10), along with eleven known analogs, were isolated from an ethanol extract of the calyx of Physalis alkekengi. The new structures were rigorously determined through comprehensive HRESIMS, 1D/2D-NMR, and X-ray diffraction analysis. Among these compounds, 1 was identified as a new 1,10-seco-neophysalin, and 2 was identified as a new 11,15-cyclo-9,10-seco-physalin characterized by an aromatic A-ring. Evaluation of the anti-inflammatory activities of the isolated compounds revealed that compound 15 displayed remarkable potency, inhibiting NO production in RAW 264.7 macrophages with an IC