Four new C-21 steroidal glycosides (1-4), named perisepiumosides F-I (1-4) together with six known steroidal glycosides (5-10) and four oligosaccharides (11-14), were isolated from the root bark of Periploca sepium. Their structures were characterized on the basis of 10 and 2D-NMR spectroscopic data as well as HR-ESI-MS analysis. The evaluation of inhibition activity against human A-549 and HepG2 cell lines indicated that compounds 2, 8,10 and 13 showed different levels of cytotoxic activities with IC50 values ranging from 0.61 to 7.86 mu M. (C) 2017 Elsevier B.V. All rights reserved.