Study On Chemistry Constituents And Activities From Paris Polyphylla

RHIZOMA PARIDIS(RP), the dry Rhizome of Paris polyphylla, which distributes widely in Sichuan, Yunnan and guizhou Province of China, is a kind of traditional Chinese herbal medicine with a long history of widely used to antipyretic, analgesia, antidote and acute nephritis, et al. Modern clinical and pharmacological research show RP have the antitumor, stop bleeding, immunity adjustment, antimicrobial pharmacological activities. However, the antitumor, antimicrobial, free radical scavenging constituents of RP are still unknown, especially the bioactivity research for pure compounds of RP. Therefore, the object of this paper is to determine the the antitumor, antimicrobial,free radical scavenging constituents of RP and the chemistry constituents of n-BuOH fraction, and this paper can be recognized as one of the database for the R&D research of new drug.The 80% ethanol extract of Paris polyphylla was partitioned successively with petroleum ether (PE), ethyl acetate (EtOAc), n-butanol (n-BuOH) and the residues of aqueous. The free radical scavenging action of each fraction was determined by using DPPH radical scavenging method in vitro. We study antibiotic function of each fraction against Staphylococcus aurous,Escherichia coli and Candida albicans. The inhibitory effect on human lung adenocarcinoma cell line A-549 of each fraction was determined. The results showed that there are other parts of constituents with anti-tumor active such as aqueous fraction besides the n-BuOH fraction from P. polyphylla. 62.5μg/ml, 125μg/ml and 250μg/ml aqueous fraction inhibition rate were 49.6%, 65.8% and 82.7% to A-549 respectively. The n-BuOH fraction has the highest antibacterial action to Candida albicans, the MIC is 5mg/ml.The aqueous fraction has the highest antibiotic function to Staphylococcus aurous and Escherichia coli, the MIC respectively were 7.5mg/ml and12.5mg/ml.The EtOAc fraction has the strongest free radical scavenging action; the IC50 was 1.1mg/ml.By using column chromatography, including silica gel chromatography, reversed phase low press preparative chromatography, four compounds were isolated from n-BuOH part of Paris polyphylla,and their structures were elucidated on the basis of spectral analyses(1H-NMR,13C-NMR). They were elucidated asβ–ecdysterone (compound1), pennogenin-3-O-α-L-rhamnopyranosyl (1→2)-β-D- glucopyranoside (compound2), pennogenin-3-O-α-L-rhamnopyranosyl (1→4)-α-L-rhamnopyranosyl (1→4)-[α-L-rhamnopyranosyl(1→2)]-β-D-glucopyranoside(compound3), compound4 is the analogue of pennogenin-3-O-α-L-rhamnopyranosyl(1→4)-[α-L-rhamnopyranosyl (1→2)]-β-D-glucopyranoside.The antifungal and antitumor activities of each compound were determined by against Saccharomyces cerevisia, Candida albicans and human lung adenocarcinoma cell line A-549. The results showed that compound2, compound3 and compound4 had significant activity against Saccharomyces cerevisia, Candida albicans; the MIC respectively were 2.5mg/ml, 0.625mg/ml and 0.625mg/ml to Saccharomyces cerevisia; the MIC respectively were 1.25mg/ml, 0.625mg/ml and 1.25mg/ml to Candida albicans. compound2, compound3 and compound4 had significant cytotoxicity on A-549 cells in a dose-dependent manner; 12.5μg/ml compound2, compound3 and compound4 inhibition rate were 82.9%, 79.0% and 56.8% to A-549 respectively, the most inhibition rates were above 85%. Compound1 had no significant antifungal and antitumor activities. The results showed showed that the glycoside exist or not and the number of glycoside in the structure of saponins strongly influenced its activities.