Studies On The Anticancer Active Constituents Of Clausena Dunniana Levl.

In the present dissertation, 142 species of Chinese medicinal herbs were tested for their anticancer activities by the use of bioassays respectively using a fungus, P oxyzae, and a mouse temperature sensitive cdc2 mutant cell line, tsFT2lO. Among the plant materials tested, 25 species caused growth inhibition or morphological changes of P oxyzae, which has been testified to be a useful model for screening of anticancer agents, and 37 species showed cell cycle inhibitory, apoptosis inducing and/or cytotoxic activities on tsFT2lO cells. Totally, 17.6% and 26.1% of total species examined are positive respectively on P. oxyzae and tsFT2lO cells in the present study. On the basis of the above result, relations and differences between two bioassays for evaluating anticancer activities were also discussed in the dissertation. During the screening, we found that the 70% aqueous alcohol extract of the plant Clausena dunniana Levl., which was collected from Mengla area of Yunnan province, showed strong anticancer activities in the two bioassays. Especially in the tsFT2lO bioassay, it showed obvious cell cycle inhibitory, apoptosis inducing and cytotoxic activities on tsFT2lO cells. C. duuniana is a tree of genus Clausena (family Rutaceae), and is used as a folk medicine to cure cancer in partial area of China. So far, a number of reports on the chemical constituents of genus Clausena had been seen and essential oil, alkaloid and coumarin were reported as major component. However, no research report had so far been seen on the chemical constituents of the title plant. So we undertook the research program on the active principles of C. duuniana and 29 carbazole alkaloids were isolated from the stem bark and branch of the title plant through bioassay-guided separation procedures. The structures of carbazole alkaloids were elucidated on the basis of the physico-chemical properties and spectra data and are showed in Fig 1. Among them compounds 2, 5, 11, 13, 26 and 28 are new carbazuole alkaloids, 24 and 25 are new natural products, and 8, 9, 12, 1549, 21, 22 and 27 are newly isolated from the genus Clausena. Incidentally, all of the compounds [?9 are 4 isolated for the first time from C. duuniana Levi.. In the course of bioassay-guided separation, from the extraction of crude extract or the separation of effective fractions to the isolation of active chemical components, each step was monitored by the bioassay of detecting the cell cycle inhibitory, apoptosis-inducing and cytotoxic activities on tsFT2lO cells. The principles, which were beneficial and effective for the separation of the active compounds, were the rule for selecting the follow isolation method during separation procedure. The separation programs were decided on the basis of the identification of the active TCL spots revealed by the micro-sample activity test. Therefore, the way for isolations could ensure the active compounds to be separated fast and accurately. Among the 29 compounds isolated, 23 of them showed their activities with different characters and intensities on tsFT2 10 bioassay. The other six compounds, which hadn抰 showed any activity on the tsFT2 10 bioassay, were also isolated and identified for their carbazole structures for which may help to discuss of the structure-activity relationships (SAR). Throughout careful analysis of the spectra of the compounds 1?9, some general rules for the carbazole alkaloids?structure elucidation were summed up. In addition, the...