| Objective: Sophorae Tonkinensis Radix Et Rhizoma,a member of Leguminous,is the dry root and rhizome of Sophorae Tonkinensis Gagnep,also known as Shan Dou Gen,Guang Dou Gen,and Yue Nan Huai,etc.The chemical constituents of Sophorae Tonkinensis Radix Et Rhizoma are mainly alkaloids,flavonoids,terpenoids and steroid compounds.Literature investigation shows that the main active component of Sophorae Tonkinensis Radix Et Rhizoma is alkaloids,but other compounds in Sophorae Tonkinensis Radix Et Rhizoma also have irreplaceable biological activities.We found that the chloroform extract(CE)of Sophorae Tonkinensis Radix Et Rhizoma is the main bioactive component of anti-nasopharyngeal cancer in our early activity screening research.Therefore,insearching for bioactive compounds,the chemical components of its CE were studied,and the components of its fraction were isolated and identified by directional tracing,so as to explore its anti-nasopharyngeal active mechanism.Methods: Powder of Sophorae Tonkinensis Radix Et Rhizoma 55 kg was soaked with 75% ethanol for three times repeatedly.The extracts were extracted with petroleum ether,chloroform respectively,and the CE was obtained 1100 g.The CE fraction of Sophorae Tonkinensis Radix Et Rhizoma was analyzed by HPLC method and the fingerprint of the anti-nasopharyngeal cancer constituents from Sophorae Tonkinensis Radix Et Rhizoma was established by HPLC.Under the direction of HPLC fingerprint,the anti-nasopharyngeal cancer constituents were targeting isolated and purified by column chromatography by repeated silica gel,sephadex LH-20 gel or ODS etc from the Sophorae Tonkinensis Radix Et Rhizoma.The structures of the isolated compounds were elucidated on the basis of spectroscopic analysis(MS,NMR etc).Results: 1.0 ml/min for liquid velocity,205 nm for detection wavelength,30℃ for temperature,20 μl for sample volume,gradient elutioned with aqueous solution(A)and acetonitrile(B).The fingerprint of the anti-nasopharyngeal cancer constituents of Sophorae Tonkinensis Radix Et Rhizoma was established.And 20 compounds were separated from the Sophorae Tonkinensis Radix Et Rhizoma and they were identified as lupeol(1),3-methyloxy maackiain(2),tonkinensisol(3),sophoranochromene(4),maackiain(5),4’,7-dihydroxy-6,8-bis(3-methyl-2-butenyl)flavanone(6),maltol(7),syringaresinol(8),tonkinochromane H(9),ononin(10),medicagol(11),caffeic acid(12),2-(2’,4’-dihidroxyphenyl)-5,6-methylenedioxy benzofuran(13),formononetin(14),soyasapogenol B(15),2-[{3-Hydroxy-2,2-dimethyl-8-(3-methyl-2-butenyl)}chroman-6-yl]-7-hydroxy-8-(3-methyl-2-butenyl)chroman-4-one(16),genistein(17),7,4’-dihydroxyflavone(18),trifolirhizin(19),trifolirhizin 6’-ethanoyl(20).Preliminary screening studies on the anti-nasopharyngeal cancer activity of the above compounds in vitro showed that the inhibition rates of compounds(2),(13)and(19)on CNE-one cells were 60.90%,94.41% and 51.82%,respectively.The inhibition rates of compounds(1),(2),(5),(13)and(16)on CNE-two cells were 90.53%,64.73%,74.76%,99.03% and 98.07%,respectively.The peaks of the compounds in the fingerprint were identified by HPLC.Conclution: 20 compounds are isolated from the CE of Sophorae Tonkinensis Radix Et Rhizoma,among them,compounds(2),(8)and(11)are isolated from this plant for the first time.The results of pharmacological experiments shows that the CE of Sophorae Tonkinensis Radix Et Rhizoma has good anti-nasopharyngeal cancer activity.The anti-nasopharyngeal cancer activity of the compounds isolated from this fraction is screened by MTT in vitro cell model,and it is found that the compounds(1),(2),(5),(13),(16)and(19)have better inhibitory activity against proliferation of nasopharyngeal cancer cells.And on this basis,HPLC fingerprint of the quality control of antinasopharyngeal cancer active fraction is preliminarily explored,which laid a foundation for the further development and research of Sophorae Tonkinensis Radix Et Rhizoma. |
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