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We have evaluated the in‐vitro and in‐vivo antitumour activity of physalin B and physalin D isolated from the aerial parts of Physalis angulata. In‐vitro, both compounds displayed considerable cytotoxicity against several cancer cell lines, showing IC50 values in the range of 0.58 to 15.18 μg mL⁻¹ for physalin B, and 0.28 to 2.43 μg mL⁻¹ for physalin D. The antitumour activity of both compounds was confirmed in‐vivo using mice bearing sarcoma 180 tumour cells. The in‐vivo antitumour activity was related to the inhibition of tumour proliferation, as observed by the reduction of Ki67 staining in tumours of treated animals. Histopathological examination of the kidney and liver showed that both organs were affected by physalin treatment, but in a reversible manner. These compounds were probably responsible for the previously described antitumour activity of ethanol extracts of P. angulata, and their identification and …