Deubiquitinating protease USP7 is a promising therapeutic target for cancer treatment,and interest in developing USP7 inhibitors has greatly increased.In the present study,we reported a series of natural pentacyclic triterpenes with USP7 inhibitory activity in vitro.Among them,both the ursane triterpenes and oleanane triterpenes were more active than the lupine triterpenes,whereas ursolic acid was the most potent with IC50 of 7.0±1.5 μmol/L.Molecular docking studies showed that ursolic acid might occupy the ubiquitin binding pocket of USP7,with the 17-carboxyl group and 3-hydroxyl group playing a vital role in the USP7-ursolic acid interaction.Using the cellular thermal shift assay,we demonstrated that ursolic acid interacted with USP7 in RPMI8226 human myeloma cells.Ursolic acid dose-dependently inhibited the proliferation of the myeloma cells with IC50 of 6.56 μmol/L,accompanied by reductions in USP7 substrates such as MDM2,UHRF1 and DNMT1.Overexpression of USP7 partially,but significantly attenuated ursolic acid-induced cell death as well as downregulation of MDM2,UHRF1 and DNMT1.In conclusion,we demonstrate for the first time that pentacyclic triterpenes represent a novel scaffold for developing USP7 inhibitors and that USP7 inhibition contributes to the anti-cancer effect of ursolic acid.