Studies On Chemical Constituents And Anti-Tumour Activity Of Active Fraction From Chloranthus Japonicus Sieb.

Chloranthus japonicus Sieb is a deciduous perennial herb, which belongs tothe family of Chloranthaceae. It has the long history to be used as traditionalChinese medicine and utilized widely to cure many diseases such as cough,rheumatalgia, amenorrhea, injury from fall, ecchymoma and painful, coughingup blood etc. due to its strong and wide biological activities. C. japonicusdistributes mainly in many provinces of China such as Helong-jiang, Jilin, Hebei,ShanXi, ShanDong, Shanxi, Gansu, etc. Although it has the abundant naturalresource, this medicinal plant has not almost been developed and exploited for along time.Our research groups have screened anti-tumor medicine from more than 20plants, and found that the gross extraction (CJ) of C. japonicus has stronganti-tumor activity. In order to clarify anti-tumor activity of CJ and confirm theactive parts, CJ was separated for four different parts named CJ-1, CJ-2, CJ-3 andCJ-4 by macroporous resin and solvent methods. The four elutions were studiedby the inhibitory ratio of the tissue of tumor-bearing mice S₁₈₀, H₂₂, EAC. At last,CJ-3 was proved the strongest activity part in them.30 compounds have been isolated from the active fraction CJ-3 by silica gel,ODS, Sephadex LH-20 column chromatography and HPLC. 27 compounds havebeen elucidated their chemical structures by chemical and spectroscopy evidence,such as UV,IR,¹H-NMR,¹³C-NMR,¹H-¹H COSY,HSQC,HMBC and NOESY,combined with ESI-MS, HR-ESI-MS. Among them there are 9 sesquiterpenes(yinxiancaoside D, chloranosides A, yinxiancaoside E, 3S, 5R, 8R- 3, 5-dihydroxy-megastigma-6, 7-dien-9-one, 3S, 5R, 8R-3, 5-dihydroxymegastigma-6, 7-dien-9-one-3-O-β-D-glucopyranoside, pisumionoside, 3S, 5R, 6R, 7E, 9S-tetrahydroxy-megastig-mane-7-en-3-O-β-D-glucopyranoside, 3S, 5R, 6S, 7E, 9S-tetrahydroxy-megastigma-ne-7-en-3-O-β-D-glucopyranoside, 3S, 5R, 6R, 7E, 9Stetrahydroxymegastigmane-7-en-3-O-and-9-O-β-D-diglucop-yranoside),10 lignanoids (7S, 8R-dihydrodehy-drodiconiferylalcohol, 7S, 8R urolignoside, 7S, 8R-dihydrodehydrodiconiferylalcohol-9-O-β-Glucopyranoside, 7S, 8R-dihydrodehy-drodiconiferylalcohol-9'-O-β- glucopyranoside, 7S, 8R-5-methoxydihydrodehydrodiconife-rylalcohol-4-O-β-glucopyranoside, 7S, 8R-4, 7, 9, 9'-tetrahydroxy-3, 3'-dimethoxy-8-O-4'-neolignan, 7S, 8R-7,9, 9'-trihydroxy-3, 3'-dimethoxy-8-O-4'-neolignan-4-O-β-D-glucopyranoside, 7S, 8R-7, 9, 9'-trihydroxy-3, 3', 5'-trimethoxy-8-O-4'-neolignan-4-O-β-D-glucopyranoside,(+)-pinoresinol-O-β-D-glueopy-ranoside, cleomiscosinC-4-O-β-glueopyranoside), 4saponin compounds (3-O-α-L-arabinoPyranosyl-hederagenin-28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glueopyranosyl-(1→6)-β-D-glucopyranosylester, 3-O-α-L-arabinopyranosyl-caulophyllogenin-28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl-ester, 3-O-β-D-glucopyranosyl-(1→2)-α-L-arabinopyranosylhedera-genin-28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glueopyranosyl-(1→6)-β-D-glucopyranosyl ester, 3-O-β-D-glucopyranosyl-(1→3)-α-L-arabinopyranosyl-hederagenin-28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glueopyranosyl ester) and 4 others (5'-hydroxyjas-mone 5'-O-β-D-glucopyranoside, androsin,β-acuminosidase, 4-methyl-2, 6-dime-thoxylphenol-1-O-β-D-glueopyranoside). In particular, there are a novel com-pound (4-methyl-2, 6-dimethoxyl-phenol-1-O-β-D-glucopyranoside) and 17 com-pounds firstly found in Chloranthus japonicus Sieb..MTT was also used to test the antitumour activity of these isolatedcompounds on Hepg-2, OV420, MCF-7 human tumour cell lines. Sesquiterpenes,dihydrobenzofuran ligins and triterpenoids showed strong antitumor activities.The structure-activities relationship indicated antitumor activity sourced fromskeleton of compouds. There are some selectivity on human tumour cells comefrom stereo-isomer and substituent group variance.This dissertation elucidated the anti-tumor material basis of C. japonicus Sieb.by chemical and pharmalogical research. It will provide important scientificfoundation for further research and the development of new drugs and promotethe exploitation of this abundant natural medicinal resource.