• The traditional Chinese medicine Centella asiatica was studied. • Seven flavonoids including two new ones were isolated. • Two new flavonol derivates exhibited potent cytotoxic activities on HepG2 and SGC-7901 cells. • Compound 2 showed higher cytotoxic activity on HepG2 cells with IC 50 value of 4.52 μM. Centella asiatica is well known as an important medicinal plant because of its various pharmacological effects. However, most investigations on C. asiatica focused on the pharmacological activity of its extract or asiatic acid. In the present work, we aimed to explore the bioactive compounds of the whole plants of C. asiatica . As a result, seven compounds including two new flavonol derivates named 4′-hydroxyl-7-methoxyl-6-prenyl-3- O - trans - p -coumaroyl-flavonol ( 1 ) and (2 R ,3 R, 2″ S )-3-furanoyl-brosimacutin E ( 2 ), and five known compounds ( 3-7 ) were isolated. Their chemical structures were elucidated on the basis of spectroscopic analyses. All the isolates were evaluated for in vitro cytotoxic effects on four human cancer cells including A549, HepG2, SGC-7901 and MCF-7. Among them, compounds 1 and 2 exhibited higher cytotoxic activities on HepG2 and SGC-7901 cells compared with 3-7 . And compound 2 showed potential cytotoxic activities on HepG2 and SGC-7901 cells with IC 50 values of 4.52 and 7.03 μM, respectively.