Studies On Bio-Active Constituents Of Four Medicinal Plants

The stem and bark of Daphne giraldii Nitsche (Thymelaeaceae), as well as the stem and bark of D. tangutica Maxim, D. retusa Hemsl., D. koreana Nakai., and Edgeworthia chrysantha Lindl., commonly called"Zu Shima", is widely distributed in Shanxi, Gansu, Ningxia, Sichuan and Qinghai provinces of China. It has been used in Chinese folk medicines to treat aches and rheumatism. In modern pharmacological investigation, it was used to treat toothache, waistache, rheumatoid arthritis, quadriplegia, thrombosis, inflammation, cancer etc. D. retusa, also called Guihua Aituotuo, belongs to the genus of Daphne, Thymelaeaceae family. Its stem and barks are used to treat injuries from fall, and in Zhejiang, it is also treat chronic infantile convulsion in folk; D. koreana belongs to the genus of Daphne and it is only distributed in Changbai Mountain. It is also called La Gencao. It is used for treatment deadlimb, thoracodynia, coronary artery disease, rheumatic arthritis, thromboangiitis obliterans. E. chrysantha Lindl. which collected in JiuJiang, Jiangxi Province in May, 2005, is also called DaJie Hua, belongs to the Edgeworthia genus. The stem and bark has been used to the treatment of injuries from falls, bruisesand faucitis as a folk medicine in China. The flowers of E. chrysantha are also used to treat some diseases in eyes as a supplement of Meng Hua. In this dissertation, we investigate the chemical constituents of the stem and bark of D. retusa Hemsl., D. koreana Nakai. and E. chrysantha Lindl. systematically, and the anti-inflammatory and analgesic activities of the crude extracts, four extractive fractions, and some pure compounds.In anti-inflammatory and analgesic activity tests of all crude extracts, four extractive fractions and three pure compounds from these three plants, the results showed that the crude extracts of D. koreana, D. retusa, and E. chrysantha significantly inhibited the ear edema induced by PX and paw edema induced by complete Freund's adjuvant. Meanwhile, they also exhibited the inhibitory effects on the NO production in LPS-induced RAW264.7 macrophages in vitro test. Besides, the crude extracts also showed a significant inhibition of writhing induced by acetic acid. We also investigated chloroform and acetoacetate fractions from D. koreana, D. retusa, and E. chrysantha showed some anti-inflammatory and analgesic activities.65 compounds were obtained from chloroform and acetic ether fractions, the active parts of three plants, by a variety of chromatography methods, and their structures were established on the basis of spectroscopic data including UV, IR, ESI-MS, 1D-NMR, 2D-NMR, physical and chemical properties as follow: DR-1: p-hydroxybenzonic acid, DR-2:β-sitosterol, DR-3: daucosterol, DR-4: daphnetin, DR-5: daphnoretin, DR-6: syringaldehyde, DR-7: 7-hydroxy-8-methoxycoumarin, DR-8: yuankanin, DR-9: 7- methoxy-8-hydroxycoumarin, DR-10: umbeliferone, DR-11: edgeworoside A, DR-12: edgeworoside C, DR-13: 4', 5-dihydroxy-3', 7-dimethoxyflavone, DR-14: daphneticin, DR-15: 5''-demethoxydaphneticin, DR-16: daphneolon, DR-17: syringin, DR-18: (?)-pinoresinol, DR-19: daphnoretin-7-O-β-D-glucoside, DR-20: genkwanin, DR-21: umbeliferone-7-O-β-D-glucoside, DR-28: [8, 8'-Bi-2H-1-benzopyran]-2, 2'-dione, 7'-(β- D-glucopyranosyloxy)-7-hydroxy-3-[(2-oxo-2H-1-benzopyran-7-yl)oxy], DK-1:β-sitosterol, DK-2 octadecyl affeate, DK-3: daphnetin, DK-4: umbeliferone, DK-5: daucosterol, DK-6: 6, 7-dihydroxycoumarin, DK-7: daphnoretin, DK-8: kaempferol, DK-9: daphneticin, DK-10: luteolin, DK-11: apigenin, DK-12: sakuranin, DK-13: genkwanin, DK-14: yuankanin, DK-16: luteolin-7-methylether-5-O-β-D-glucoside, DK-17: daphnoretin-7-O-β-D-glucoside, DK-18: isodaphneticin, DK-19: 5′′-demethoxy- daphneticin, DK-20: syringin, DK-25: daphnin, JX-4: edgeworin, JX-6: daucosterol, JX-7: umbeliferone, JX-8: daphnoretin, JX-9: edgeworoside A, JX-10: 5-hydroxy-7- methoxycoumarin-8-O-β-D-glucoside, JX-11: apigenin, JX-12: edgeworoside C, JX-13: edgeworoside B, JX-14: isodaphnoretin, JX-15: 2,6-dimethoxyquinone, JX-16: umbeliferone-7-O-β-D-glucoside, JX-17: syringin, JX-18: daphnoretin-7-O-β-D- glucoside, JX-19: daphneticin, JX-20: 6-hydroxyl-odesmethoxyrutarensin, JX-21: 8-(4-(1, 3-benzenediol)propionic acid methylester)-coumarin-7-O-β-D-glucoside, and JX-23: triumbellin. JX-4, JX-9, DR-5, DR-16 and DK-3 displayed significant inhibition in ear edema induced by PX and paw edema induced by complete Freund′s adjuvant with hydrocortisone and examethasone as a positive control. JX-4, JX-9, JX-12, DR-5, DR-16, DK-4, DK-9 also showed a significant analgesic activity in writhing induced by acetic acid.The chemical investigation of other extracts of the three plants resulted in the isolation of 21 compounds and 14 structures of them were determined as following also counted on the spectral analysis and physic-chemical properties: DR-22: daphnodorin A, DR-23: daphnodorin B, DR-24: daphnodorin C, DR-25: daphnodorin D1, DR-26: daphnodorin E, DR-27: daphnodorin F, DK-21: (?)-pinoresinol, DK-22: lariciresinal, DK-23: (?)-pinoresinol-4-O-β-D-glucopyranoside, DK-24: lariciresinol-4-O-β-D- glucoside, JX-1: glycerol monostearol acid, JX-2: p-hydroxybenzoic acid, JX-3:β-sitosterol, and JX-5: inositol.Among all the compounds mentioned above, JX-20 and JX-21 were new compounds and DR-4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 28 , DK-2, 3, 4, 5, 7, 8, 10, 11, 13, 14, 16, 17, 18, 19, 21, 22, 23, 24, 25, JX-5, 11, 14, 15, 16, 17, 18, 19, and 23 were isolated from these plants for the first time.Inula salsoloides (Turc.) Ostenf. belongs to the genus of Inula, Compositae. Its flowers and the entire herbs before flower can be used fever releasing and uragoga, the plant is also a pricinpal agent to treat angeitis. It has a wide distribution in Xinjiang, Inner Mongolia, Shan′xi, Shanxi and Hebei. The chemical investigation of this plant resulted in the isolation of 32 compounds by a variety of chromatography methods, and 24 compounds were established on the basis of spectroscopic data including UV, IR, ESI-MS, 1D-NMR, 2D-NMR, physical and chemical properties, as follows: IS-1:β-sitosterol, IS-2:cis-1-p-menthene-3, 6-diol,IS-3: n-dodecane, IS-4: p-hydroxybenzonic acid, IS-5: ethyl hydroxybenzoate, IS-7: 3', 4', 7-trihydroxy-5- methoxyflavone, IS-8: stigmasterol, IS-9: 1-Cyclodecene-2, 5, 6, 9-tetrol, 1, 8-dimethyl-7-(1-methylethyl)-, [(2α, 5β, 6α, 7α, 8β, 9α)], IS-10: inulasalsolide B, IS-11: isoalantolactone, IS-12: kaempferol, IS-13: inulasalsolide, IS-14: florilenalin, IS-15: acacetin, IS-16: quercetin, IS-17: physcion, IS-18: apigenin, IS-20: 8, 9, 10-trihydroxythymol, IS-21: 9, 10-dihydroxythymol, IS-23: 4', 7-dihydroxyflavone, IS-24: luteolin, IS-25: eupatolide, IS-26: 4α, 5β-epoxyeupatolide, IS-30: 2-(3, 5-dihydroxy- phenyl)-5, 7-dihydroxyflavone, and IS-32: 4', 6, 7-trihydroxyflavone.Among all the compounds mentioned above, IS-9 and IS-10 were new compounds and IS-1, 4, 5, 7, 8, 12, 14, 15, 17, 18, 20, 21, 23, 24, 26, 30, and 32 were isolated from this plant for the first time.Conclusion: 84 compounds in total were obtained from three series plant as Zu Shima, including two new compounds and 52 compounds for the first time. 24 compounds, including two new compounds and 16 compounds for the first time were obtained from I. salsoloides (Turc.) Ostenf.. The 75% ethanol extractions, chloroform and acetic ether fractions of three theries of plants as Zu Shima showed significant anti-inflammatory and analgesic effects and on the basic of bioassay, JX-4, JX-9, DR-5, DR-16 and DK-3 showed anti-inflammatory activities, JX-4, JX-9, JX-12, DR-5, DR-16, DK-4, DK-9 and DK-25 exhibited analgesic acitivities.