Thirteen compounds were obtained from the ethanol extract of the roots and stems of Cynanchum paniculatum by applying various chromatographic techniques.Their structures were elucidated and identified as glaucogenin A (1),glaucogenin A 3-O-β-D-cymaropyranoside (2),N-(N-benzoyl-S-phenylalaninyl)-S-phenylalaninol acetate (3),N-(N-benzoyl-S-phenylalaninyl)-S-phenylalaninol benzoate (4),androst-4,6-dien-3,17-dione (5),20-hydroxy-4,6-diene-gestrol-3-one (6),5,7-dihydroxy-6-methoxyflavone (7),5,7-dihydroxy-8-methoxyflavone (8),5-(hydroxymethyl)-1-isopentyl-1H-pyrrole-2-carbaldehyde (9),mono-terpenes (10),4-hydroxy-3,5-dimethoxybenzoic acid (11),pavonisol (12),4-hydroxy-3,5-dimethoxy-benzoic acid (13),respectively,based on the spectral data.Among them,compounds 2-5 and 7-10 were obtained from the fruits of C. paniculatum for the first time.The results showed that compounds 1-4 and 9 have anti-TMV activity,through in vitro disc method and inactivation effect tested.The protection effect test showed that compound 4 has anti-TMV activity.It was found that compound 3,4 and 9 have anti-TMV activity for the first time.