Study On Chemical Constituents And Anti-tumor Activities Of Bufonis Venenum

Bufonis Venenum is the dry ear gland secretions of Bufonidae animal Bufo Bufo gargarizans Cantor or Bufo melanostictus Schneider.With the function of detoxification,relieving pain,resuscitation,Bufonis Venenum contains bufadienolide,non cardiac sterols,indole derivatives,other various types of chemical composition and so on.In order to elucidate the chemical constituents and pharmacological action of Bufonis Venenum.The chemical constituents of the acetone,methanol and water extracts were separated and anti-tumor activity of these constituents was screened.Twenty compounds were isolated and purified by the MCI column chromatography,ODS column chromatography,Sephadex LH-20 column chromatography,silica gel column chromatography,C18 and C8 reversed phase column chromatography,pre-HPLC and HPLC.According to their physical and chemical properties,structures were identified using UV?ESI-MS,HR-ESI-MS,'H-NMR,13C-NMR,DEPT,HSQC,HMBC,1H-1-NOESY spectroscopy data and detemined as cinobufotalin(1),bufalin(2),telocinobufagin(3),arenobufagin(4),bufotalin(5),bufotenine C(6),argentinogenin(7),suberic acid(8),desacetylbufatalin(9),16-desacetyl-19-Oxocinobufotain(10),?-bufarenogin(11),hellebrigenin(12),cholesterol(13),5-Hydroxy-N-acetyltryptamine(14),hippophamide(15),isobufalin methyl ester(16),n-pentadecanoic acid(17),?-sitosterol(18),1,1-Dimethyl-6-hydroxy-1,2,3,4-tetrahydro-1H-?-carboline(19),bufothionin(20).Among them compounds 16,19 were the new natural product,compounds 14,15 were isolated for the first time from Bufonis Venenum.MTT method was adopted on some compounds of Bufonis Venenum on human non-small cell lung cancer lines(A549)and human lung cancer cells(PC-9),human myeloma cells(MCF-7),human myeloma cells(OPM2)and human myeloma cells(ARP1)and the growth inhibition rate of them were measured.The results showed that compounds on one or more tumor cells showed significant inhibition.