| This thesis comprises five chapters altogether. The first chapter presents a review on the chemical constituents and bioactivities of the plants of the Tripterygium genus. The second chapter elaborates the studies on the chemical constituents of T. hypoglaucum (Levl.) Hutch. In the third chapter, bioactivities of the separated compounds from T. hypoglaucum are evaluated. The Chromatographic Fingerprints of some Tripterygium plants and tablets were investigated by HPLC method in the fourth chapter. At last, a conclusion of this dissertation is made in the fifth chapter.Plant toxin is a significant phenomenon, there are aboundant abstruse biological informations in it. In many traditional medical plant, the toxin are the active components, they may benefit for some difficult miscellaneous diseases such as cancer with appropriate method and dose. Therefore, we investigated the chemical constituents and biological activity of Tripterygium Hypoglaucum for the potent precursor compounds.The plants of the genus Tripterygium (Celastraceae), commonly known as thunder god vine, are perennial vine-like plant indigenous to Changjiang valley, southwestern and northeastern China, including Tripterygium wilfordii, T. Hypoglaucum and T. Regelii. It has a long history of use in traditional Chinese Medicine (TCM) for treating swelling, inflammation, tumor, rheumatoid arthritis (RA) and a variety of skin conditions, as well as insecticide. Although the research data regarding T. wilfordii are extensive, fewer reports on T. hypoglaucum are available, so we have investigated it.First of all, this thesis reported the investigation on the chemical constituents and bioactivities of T. hypoglaucum (Levl.) Hutch. By the means of many different chromatographic methods (silica gel, Sephadex LH-20, RP-18, Toyopearl, ODS, etc.) and spectroscopic evidences (NMR, MS, UV, etc.),45 compounds were isolated and identified from T. hypoglaucum medicinal plants, including 5 new ones: 19-O-β-D-glucopyranosyl-labda-8(17),14-dien-13-ol (1),3-oxo-14,15-dihydroxy -abieta-8,11,13-trien-19-ol (2), isopimara-8(14),15-diene-11β,19-diol (3), isopimara-8(14),15-diene-12α,19-diol (4), and 2a,16α-hydroxy-ent-kauran-19,20-olide (5); 2 new natural products:20β-β-D-glucopregn-4-en-3-one (32), 20a-β-D-glucopregn-4-en-3-one (33); 7 known diterpenes:triptolide (6), 2-epitripdiolide (7), wilforol F (8), triptobenzene A (9), isopimara-8(14), 15-diene-17,19-diol (10), tripterinin (11), and tripterifordin (12); 12 known oleanane-type triterpenes; 7 lignans; 3 steroids and 9 other type compounds. Compound 1 was a labdane diterpene glycoside, which was an additional β-D-glucose substituent on C-19; 2 was an abietane diterpene, which kind of diterpenoid were very popular in Tripterygium, there was a new substituent of OH on C-15; compounds 3 and 4 were isopimaranes, there were the substituent of OH on C-11,19 and C-12,19, respectively; 5 was an ent -kaurane diterpene, the OH substituent on C-2 was different from the known compounds.Most separated compounds were evaluated for anti-cancer and anti-inflammatory activities in vitro. The cytotoxicity evaluation used five tumor cell lines A-549, DU-145, KB, KBvin, and MDA-MB-231, and paclitaxel was the positive control. Compounds 6 (triptolide) and 7 (2-epitripdiolide) showed potent cytotoxic activity, even were equipotent against the parental KB and drug-resistant KBvin tumor cell lines, while the corresponding activity of paclitaxel was greatly reduced against the latter cell line. Compounds 25,32 and 34 also showed weak anti-tumor activity. Anti-inflammatory activity was evaluated through superoxide anion generation and elastase release. Compounds 13,14 and 15 showed obvious anti-inflammatory activity, and they were all oleanane-type triterpenes, which suggested that the oleanane triterpenes may play an important role in the anti-inflammatory activity of T. hypoglaucum. The bioactivitiy results further suggested that although terpenoids were the most important bioactivity ingradients of T. hypoglaucum, the other type compounds, such as lignans and steroids, also have the synergistic effect.Moreover, the Chromatographic Fingerprints of 3 Tripterygium plants and 3 tablets were investigated by HPLC method, and some components separated from T. hypoglaucum were fingered out in the chromatogram. The fingerprints were determined by Kromasil C18 column (4.6 mm × 25 cm,5μm) with a gradient mobile phase consisting of acetonitrile and water at a flow rate of 1 mL/min. The methods were stable, reliable and repeatable. The fingerprints of 3 Tripterygium plants indicated the chromatogram of T, wilfordii and T. hypoglaucum from Yunnan were similar, while that of T. regelii was more different from them, which may be caused by the different medical origin and different parts. The fingerprints of 3 Tripterygium tablets showed some similarity, it may due to the similar drug parts and similar process technology. Finally, the contents of triptolide in 3 Tripterygium plants were determined by HPLC, and the results were 7.38,7.16 and 9.15 μg/g, respectively.The above results, together with the literatures, showed the chemical components of different part of T. hypoglaucum (root bark, xylem, stems and leaves) were very distinct, the triptolide and most terpenoids in stems and leaves were less than those of xylem and root bark. Sesquiterpene alkaloids, which separated from root of T. hypoglaucum, were very few in leaves and stems, and there were more triterpenoids and high polar components such as lignan and phenolic compounds in leaves and stems of T. hypoglaucum in our study. There were a variety of deterpenoid, such as abietane, ent-kaurane, isopimarane, labdane, in T. hypoglaucum, which dominated the most important biological activity, the triterpenoids in it showed more anti-inflammatory activity. |
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