Pellinus lonicerinus is a mushroom that belongs to the genus Sanghuangporus and it is commonly used in Hubei and other places,which is also commonly used by Tujia ethnic minority.To investigate the chemical constituents of the culture of P. lonicerinus and their cytotoxicity,the constituents of the fermentation liquid and mycelium from P. lonicerinus  were separated and purified by chromatographic techniques,including column chromatography of silica gel,Sephadex LH-20 and semi-preparative HPLC.Their structures were mainly elucidated by NMR and MS spectroscopic techniques.Ten compounds were isolated and purified.They were identified as cyclo (L-Val-L-Pro) (1),cyclo (L-Leu-L-Pro) (2),cyclo (L-Ile-L-Pro) (3),cyclo (L-Ala-L-Pro) (4),ergosterol (5),ergostero-7,22-diene-3,6-diketone (6),ergosterol peroxide (7),phenylacetic acid (8),P-hydroxy benzaldenhyde (9) and 4-hydroxyphenylacetic acid (10).Except compounds 5,6 and 7,the other seven compounds were isolated from this fungus for the first time.Compounds 1-10 were tested for their cytotoxicities against human tumor cell lines HGC-27,HeLa,MCF-7,RL95-2,and compounds 1-4 exhibited inhibitory effects on HGC-27 and RL95-2 cell lines.Especially,the most promising compound 3 inhibited the proliferation of HGC-27 and RL95-2 cell lines with IC₅₀ values of 6.69 μM and 12.81 μM,respectively.