The invention discloses 3,16-androstenedione compounds or pharmaceutically acceptable salts thereof and application of the 3,16-androstenedione compounds or pharmaceutically acceptable salts in preparation of immunosuppressive and anti-tumor drugs. The 3,16-androstenedione compounds are represented in a following formula, are obtained by performing extraction and separation from Epigynum, and havefollowing structural formula, wherein R1 is selected from hydrogen, hydroxyl groups, methyl groups, halogen, carbonyl groups, aliphatic hydrocarbon groups and fatty amino groups, and R2 is selected from hydrogen, hydroxyl groups, methyl groups, halogen, aliphatic hydrocarbon groups and fatty amino groups. It is proven through experiments that the compounds have strong immunosuppressive and anti-tumor activities, and lead compounds are provided for development of immunosuppressants and anti-tumor preparations by the compounds, so that development and utilization of plant medicinal resources are facilitated.