Twelve compounds were isolated from the stem of Physalis pubescens L., including two categories: withanolides (1-9) and amide alkaloids (10-12). This is the first report on the occurrence of compounds 3, 11 and 12 in Physalis pubescens L. These compounds were elucidated using spectroscopic methods and by comparing their data to those reported in the literature. On the basis of chemical research, the chemotaxonomic significance of the isolated compounds was discussed.