In the present study, three previously undescribed steroidal glycoalkaloids (compounds 1–3) were isolated from Solanum lyratum. Their structures were elucidated based on comprehensive spectroscopic data. Their anti-angiogenesis and anti-metastatic activities were evaluated by MTT and wound-healing assays, respectively. Tumor-derived vascular endothelial cells (TdECs), obtained by co-culture of A549 and human umbilical vein endothelial cells (HUVECs), were treated with compounds 1–3. Results showed that compounds 1–3 significantly inhibited the migration of TdECs at 25 μM despite the weak cytotoxic activities, which indicated that the compounds exerted anti-tumor activities by inhibiting metastasis, rather than directly inhibiting the proliferation of TdECs. © Journal of Chinese Pharmaceutical Sciences, School of Pharmaceutical Sciences, Peking University