Studies On The Chemical Constitutents And Bioactivities Of Three Plants

The main content of this dissertation focuses on the studies on the chemical constituents and bioactivities of three plants, incluinding Uraria sinensis, Solanum lyratum, Acorus tatarinowii. On one hand, a total of95compounds of these plants were isolated by the solvent extraction, silica gel, and reversed-phase silica gel, gel, polyamide column chromatographies and semi-preparative HPLC and so on.80compounds were identified by the physical and chemical properties and various spectroscopic methods including NMR, MS, IR, UV, CD spectrum etc. The structural types of these compounds were involved in terpenoids, lignans, flavonoids, alkaloids, phenolic acid derivatives and so on. Among them,3new compounds were discovered, comprising a new sesquilignan glucoside from Uraria sinensis, a new spiroaminoketal alkaloid and a natural spiroaminoketal alkaloid from Solanum lyratum, a new permethylated benzofuran lignin from Acorus tatarinowii. On the other hand, Uraria sinensis was screened the main anti-rheumatoid arthritis (RA) active fraction in vivo. The cytotoxic activities of all new compounds and steroidal alkaloids were evaluated in vitro. This dissertation consists of four chapters.Chapter1Study on the screening active fractions in anti-RA and the chemical constituents of Uraria sinensisUraria sinensis Desv.(Hemsl.), a suffruticose sparingly branched perennial herb, is exclusively distributed in the Hubei, Sichuan, Guizhou, Yunnan, Shanxi, Gansu provinces of China. Traditionally, some species of this genus are used as folk medicine with the anti-inflammatory and analgesic activities or as the treatment of chills, fever, and stomachalgia. There are many investigations on the phytochemical and bioactivities of the other plants of Uraria genus in the literatures. However, Uraria sinensis has not been reported so far. As part of our ongoing search for bioactive natural compounds from the genus Uraria, we first studied on the screening active fractions in anti-RA and the chemical constituents of Uraria sinensis.The95%EtOH extract of the whole plant of Uraria sinensis was successively partitioned with petroleum ether, ethyl acetate (EE), n-butyl alcohol and water (WE). The total flavones (TF) were extracted by polyamide1010. The model of adjuvant arthritis was induced by injection of Freund’s Complete Adjuvant (FCA). Secondary paw swelling of AA rats was measured with volume meter and polyarthritis index were scored. The splenocyte proliferation, production were assayed by cell proliferation assay. After making model, mice were administered orally with different doses of EE, WE and TF fractions. The results showed that the dose2(10mg/kg),3(20mg/kg),4(100mg/kg) group of TF continued three weeks can improve obviously the symptoms of rats with rheumatoid arthritis. And dose2(40mg/kg),3(80mg/kg),4(160mg/kg) of EE and dose3of TF can decrease thymus index and spleen Index. In a conclusion, two active fractions of EE and TF are benefit to relieve the symptoms of rheumatism.Moreover, in our chemical study,46compouds were isolated from the EtOH ex traction of Uraria sinensis and41compouds were elucidated. There are including8sesquiterpenoids:(6S,7E)-6,10-dihydroxy-4,7-megastigmadien-3,9-dione-10-O-β-D-gluc opyranoside (01),3-hydroxy-5,6-epoxy-β-ionol-9-O-β-D-glucopyranoside (02),(6S,7E,9S)-6,9,10-trihydroxy-4,7-megastigmadien-3-one-10-O-β-D-glucopyranoside (03),(6S,7E,9S)-6,9,10-trihydroxy-4,7-megastigmadien-3-one-9-O-β-D-glucopyranoside (04), S-(+)-3-hydroxy-β-ionone (05),(3S,4R)-3,4-dihydroxy-β-ionone (06),5,6-epoxy-3,4-dihydroxy-7-megastigmen-9-one (07),(6S,7E,9S)-6,9,10-trihydroxy-4,7-megastigmadien-3-one (08);2triterpenoids:ginsenoside-Rgi (09), oleanolic acid3-O-β-D-glucopyranoside (10);7flavonoids:vitexin-2"-O-glucoside (11),5,4’-dihydroxy-3’-methoxyflavanone-7-(6"-O-β-L-rhamnopyranosyl)-β-D-glucopyranoside (12), apigenin (13), quercetin (14), kaemp ferol (15), isoferreirin (16), naringenin (17);5lingams:(7"S,8"R)-(-)-4,4",9,9’,9"-pent ahydroxy-3,3’,3"-trimethoxy-4’,8"-oxy-8,8’-sesquineolignan-7"-O-β-D-glucopyranoside (18),(-)-syringaresinol (19),(-)-medioresinol (20),(-)-(7R,7’R,7"S,8S,8’S,8"S)-4’,4"-dihy droxy-3,3’,3",5,5’-pentamethoxy-7,9’:7’,9-diepoxy-4)8"-oxy-8，8’-sesquineolignan-7",9"-dio1(21),(+)-(7R,7’R,7"R,7"’R,8S,8’S,8"S,8"’S)-4",4"’-dihydroxy-3,3’,3",3’’’,5,5’-hexamethox y-7,9’:7’,9-diepoxy-4,8":4’,8"’-bisoxy-8,8’-dineolignan-7",7’",9",9’"-tetraol (22);7glycos ides:bergenin (23), D-galacitol (24), xylitol (25), arbutin (26), uridine (27),β-D-Fru c-2-1-α-D-Glc (28),β-Methyl-D-glcopyranoside (29);5phenolic acid compounds:ben zoic acid (30),4-anisaldehyde (31),3,4-dihydroxybenzoic acid (32),4-hydroxybenzoi c acid (33), hordenine (34);5fatty acids:1-O-β-D-glucopyranosyl-(2S,3S,4R,8E)-2-[(2’R)-2’-hydroxytetracosanamino]-8-octadecene-1,3,4-triol (35), palmitic acid (36), stea ric acid (37), lignocericacid (38), tetracosanoic-2,3-dihydroxypropy lester (39);2ster oids:β-sitosterol (40), daucosterol (41). COM.18was a new compound. All compo unds were obtained from the plant for the first time.Chapter2Study on the alkaloids and cytotoxic activity of Solanum lyratumSolanum lyratum Thunb, a perennial herbal liana, is widely distributed in the Jiangsu, Zhejiang, and Anhui provinces of China. It has been used for the treatment of febrifuge, diarrhea, eye disease and cancer as a folk remedy in China for years. The alkaloids from Solanum lyratum are abundant and have moderate bioactivities, which were reported on many literatures. On the basis of this, the research on the alkaloids and cytotoxic activity of Solanum lyratum was carried on.The total alkaloids of ethyl acetate extraction (ETA) were obtained by ethanol acid-base precipitation.24compounds were isolated from ETA by means of repeated silica gel column chromatographies and HPLC purification, and20of them were identified. There are including8steroidal alkaloids:5a-solanidane-3β,16-diol (BY-01),7-oxosolasodine (BY-02),(22R,25R)-3β-hydroxy-22a-N-spirosol-5-ene-7-one (BY-03),4-tomatiden-3-one (BY-04),1,4-solasodadien-3-one (BY-05), soladulcidine (BY-06), tomatidenol (BY-07), solasodine (BY-08);6amides:N-trans-femloyl butyric acid (BY-09), N-cis-femloyl octopamine (BY-10), N-trans-femloyl octopamine (BY-11), N-cis-femloyl tyramine (BY-12), N-trans-femloyl tyramine (BY-13), N-trans-femloyl-3-methyldopamine (BY-14);2phenylpropanoids:dihydroconiferyl ferulate (BY-15), Dihydrosinapyl ferulate (BY-16);3steroids:24a-methylcholest-7,22-dien-3β5αa,6β-triol (BY-17), daucosterol (BY-18), diosgenin (BY-19). BY-03was a new compound. BY-01,02,03,09,10,11,12,13,15,16,17were obtained from the plant for the first time.Additionally, five tumor cells, leukemia HL-60, lung cancer A-549, of SMMC-7721, breast cancer MCF-7and colon cancer SW480and a normal lung epithelial cell BEAS-2B were selected to evaluate the steroidal alkaloids cytotoxic activity in vitro. The bioassay results revealed that BY-01showed moderate cytotoxic activities against the five selected tumor cells.Chapter3Study on the chemical constituents of Acorus tatarinowiiAcorus tatarinowii, a perennial herbal, is mainly distributed to the areas of the S. Yangzi river, especially of Sichuan, Zhejiang, Jiangsu. The rhizome of Acorus tatarinowii is a well-known traditional Chinese medicine for treating central nervous system-related disorders. In recent years, a series of novel lingnans which have neuroprotective and anti-tumor activities from Acorus tatarinowii have arose great interests of us. Therefore, we have been studying on the chemical constituents of the radix of Acorus tatarinowii so as to find more active constituents.Repeated extensive column chromatographies on the ethyl acetate extract of the rhizome of Acorus tatarinowii had resulted in24compounds.20compounds were identified as follows,5lingams:2R,3S-dihydro-2-(4-hydroxy-3-methoxyphenyl)-3-hydr oxy-methyl-7-methoxybenzofuran-5-carboxylic acid (SCP-11),(+)-syringaresinol (SCP-13),(7R,7’S,8R,8’R)-3’,4,4’-trimethoxy-7,7’-epoxyligne-3,9,9’-triol (SCP-17),(7R,8R)-7,8-dihydro-1’-(3-hydroxy-propyl)-5’-methoxy-7-(3,4-dimethoxyphenyl)-9-methylbenzofuran (SCP-19),(2R)-1,1-di(2,4,5-trimethoxyphenyl)propan-2-ol (SCP-20);8amides:1-O-β-D-glucopyranosyl-(2S,3S,4R)-2-[(2’S)-2’-hydroxytetracosanamino]-octadecane-1,3,4-triol (SCP-05),1-O-β-D-glucopyranosyl-(2S,3S,8Z)-2-[(2’R,3’E)-2’-hydroxy-3-hexadecenamide]-8-octadecene-1,3-diol (SCP-06),1-O-β-D-glucopyranosyl-(2S,3S,4E,8Z)-2-[(2’R)-2’-hydr oxytetracosanamino]-4,8-octadecadien-1,3-diol (SCP-07),1-O-β-D-glucopyranosyl-(2SS,3S,4R)-2-[(7’Z)-hexadecenamide]-octadecane-1,3,4-triol (SCP-08),1-0-β-D-glucopyranos yl-(2S,3S’,4R,8Z)-2-(tetracosanamino)-8-octadecene-1,3,4-triol (SCP-09), N-trans-p-coum aroyl tyramine (SCP-10), N-cis-p-coumaroyl tyramine (SCP-12),1-H-4-amino-2-oxo-1,2-dihydroquinoline (SCP-14);6phenylpropanoids:asarylaldehyde (SCP-01), a-asaro ne (SCP-02),β-asarone (SCP-03),1-(2,4,5)-trimethoxyphenyl-propane2-1,2-dione (SC P-04),(S)-3-hydroxy-3-(4’-methoxyphenyl)-propanoic acid (SCP-16),(S)-2-(3,4-dimeth oxyphenyl)propanal (SCP-18); a quinonoid:1,6,8-trihydroxy-3-carboxy-anthraquinon e (SCP-15). SCP-19was a new compound, and was evaluated for the cytotoxic acti vities against five human cancer cell lines. However, the bioassay results revealed th at SCP-19has no in vitro cytotoxicities against all five cancer cell lines (IC50>40μM).Chapter4ReviewsIn order to further develop and utilize of these plants, we provided3reviews on the progress of the chemical and pharmacological study of genus uraria plants, Solanum lyratum and Acorus tatarinowii.