Study On The Alkaloids From The Zanthoxylum Nitidium In Yunnan Province

The paper mainly in the roots and leaves of Yunnan,Mengla County,Xishuangbanna Prefecture of Zanthoxylum nitidum（Roxb.）DC.as the research object.There are chemical composition from the ethanol extraction part of Zanthoxylum nitidum from the roots and leaves were isolated,purified and identified by modern separation techniques,the chemical composition of these compounds were characterized through UV,ESI-MS,1D,2D NMR,physical constants determination and thin layer chromatography（TLC）.At the same time,the antitumor cells experiments in vitro were evaluated that the proliferation inhibitory effect of the compounds on three cell lines,including K562,HEL and WM9.12 compounds from the roots and 19 compounds from the leaves of Zanthoxylum nitidum（Roxb.）DC.were isolated and identified.They were（+）-9′-O-transferuloyl-5,5′-dimethoxylaricriresinol（1）,8-（3′-oxobut-1′-en-1′-yl）-5,7-trimethoxy-coumarin（2）,5,7,8-trimethoxy-coumarin（3）,5-（3’,3’-dimethyl-2’-butenyloxy）-7,8-dimethoxy-coumarin（4）,2-（5-methoxy-2-methyl-1H-indol-3-yl）methyl acetate（5）,6-acetoethyl-dihydrochleletrythrine（6）,6-acetonyldi-hydrochelerythrine（7）,bocconoline（8）,decarine（9）,zanthoxyline（10）,O-methylzanthoxyline（11）,rhoifoline B（12）,N-nornitidine（13）,nitidine（14）,5-hydroxy-7,8-dimethoxy-coumarin（15）,4-hydroxyl-7,8-dimethoxy-furoquinoline（16）,dictamnine（17）,γ-fagarine（18）,skimmianine（19）,dihydrochelerythrine（20）,R-（+）-platydesmine（21）,liriodenine（22）,espectively.Among them,the structures of eight compounds（1,2,4,5,6,11,15 and 16）were isolated from Z.nitidium for the first,including four compounds（4-5 and 16）confirmed and attributed for the first time by UV-visible spectroscopy,1D and 2D NMR and HR-ESI-MS.On the basis of chemical work,the 1-22 compounds was screened for anti leukemia and melanoma activity in vitro.Of the assayed compounds,compounds 8,14,20 and 22 showed the potent inhibitory activities against HEL cells with IC₅₀values of 3.59,6.68 and 15.95μmol·L^-1,compounds 6,8,12,14,20 and 22 showed the good inhibitory activities against K562 cells with IC₅₀values of 15.53,10.50,17.53,0.03,14.56 and 12.37μmol·L^-1,and compounds 6,14 and 22 showed inhibitory activities against WM9 cells with IC₅₀values of 4.80,0.88 and 13.33μmol·L^-1,these data show that compounds 8,14,20 and 22 had a potent proliferation inhibitory,which was the most of the alkaloids（especially benphenanthridine）.To the further toxicity experiments showed that the inhibiton rates of compounds1,4,6,7,8,12,20 and 22 were less than 30%at concentrations of 10 and 20μmol·L^-1,among the four compounds（8,14,20 and 22）with the potent proliferation inhibitory,inhibition rate of compounds 8,20 and 22 were below 0%,but inhibition rate of compounds 14 was greater than 50%.The data suggest that the compounds（8,20 and 22）may be a potential low-toxic anti-leukemia drug.In addition,to further confirm the effects of compounds 8,14 and 22 with different structures on cell cycle and apoptosis of two tumor cells（K562 and HEL）.The data indicated that compounds 8 and 14 caused obvious S phase arrest and induced cell apoptosis on two cells（K562 and HEL）,but compound 22 induced only cell apoptosis of two cells.In addition,the cell cycle and apoptosis on WM9 cells assays were investigated,which indicated that compound 14 caused obvious S phase arrest and induced cell apoptosis on WM9 cells,but compound 22 induced only cell apoptosis on WM9 cells.These results suggested that the benphenanthridine from Z.nitidium provides an important experimental basis for the development of high efficient,low-toxic and selective anti-leukemia candidate drug.