Triterpenoid saponins were extracted from of Pseuderanthemum palatiferum (Nees) Radlk. dry leaf powder by using enzyme‐assisted extraction method. Effects of single factors on total saponins content (TSC) were investigated. Then, a full factorial design with two levels was conducted to screen the main factors affecting TSC. Two main factors as extraction temperature and time were determined. Response Surface Methodology with central composite design was used to study simultaneous effects of these factors on TSC. Results showed that the optimum conditions were 54.9 °C and 80.8 min. The extract under these conditions was tested with bioactivities. The extract had the haemolytic index of 256.74 mL, anti‐inflammatory activity with IC20 of 0.56 μg/mL, anti‐oxidation activity with IC50 values of DPPH, ABTS, and hydroxyl of 61.47, 1.42, and 30.33 μg/mL, respectively, and cytotoxicity on cancer cell line HepG2. Accordingly, triterpenoid saponins from Pseuderanthemum palatiferum (Nees) Radlk. could be considered as a valuable constituent.