Studies On Chemical Constituents Of Andrographis Paniculata Nees. And Apium Graveolens L.

Chuanxinlian,the dried aerial parts of Andrographis paniculata(Burm.f.)Nees.is a Chinese herbal medicine used as an anti-inflammatory,antiviral and antipyretic drug for treatment of fever,cold,laryngitis and diarrhea.And it was specified in the Pharmacopoeia of the P.R.China(2005)as an anti-virus and anti-bacterial remedy.The chemical constituents from n-BuOH and water portions of 85%EtOH extract of the aerial parts of this plant was investigated in this thesis and led to the isolation of 27 compounds.On the basis of chemical evidences and spectral analysis,25 compounds of them were identified,and they are 14-deoxy-11,12-didehydroandrographolide(1),andrographolide (2),neoandrographolide(3),14-deoxy andrographolide(4),14-deoxyandrographiside(5), 14-deoxy-11,12-didehydroandrographiside(6),3-O-β-D-glucopyranosyl-14-deoxy-andrographiside (7),3-0-P-D-glucopyranosyl-14-deoxy-11,12-didehydroandrographiside(8), isoandrographolide(9),andrographiside(10),5,4'-dihydroxy-7-O-β-D-pyranglucuronate butyl ester(11),andrographidine C(12),apigenin-7-O-β-D-glucopyranoside(13), 5,4'-dihydroxy-7-methoxy-8-O-β-D-glucopyranosyloxyflavone(14),7,8-dimethoxy-2'-hydroxy-5-O-β-D-glucopyranosyloxyflavone(15),6-C-β-D-glucosyl-8-C-β-D-galactosyl-apigenin (16),6,8-di-C-β-D-glucosyl-chrysin(17),apigenin-7-O-β-D-glucuronide(18), 8-acetyl-harpagide(19),procumbide(20),teuhircoside(21),2,6-dimethoxyl-p-hydroxybenzoyl-1-O-β-D-glucoside(22),roseoside(23),citroside B(24),oxalic acid(25), respectively.Among these twenty-seven compounds,ten are diterpenoid lactones,eight are flavones,three are iridoids,two are megastigamane glycosides,one is a phenol glycoside and one is an organic acid.And among them,7,8 and 15 are new compounds,11,16,17,18,21, 22,23,24 and 25 were isolated for the first time from Andrographis genus.These results revealed the megastigamane glycosides and phenoglycosides were isolated for the first time from Andrographis genus,which were.important supplement to the chemical constituents of Andrographis paniculata.Apium graveolens L.,the fresh whole plant of Apium graveolens L.in Umbelliferae.,is a normal vegetable to people.It is effective on epilepsy,hyperlipemia,and hypertension;it also can protect the brain cells and enhance memory.As an ordinary vegetable in daily life, celery has a variety of pharmacological activity,and received increased attention on its chemical constituents.Previous studies were focused on celery seeds but not its fresh herb of the whole plant.Therefore,in this thesis,the chemical constituents of the EtOAc,n-BuOH and water extracts from 70%EtOH extract of the fresh herb of Apium graveolens L.were studied.43 constituents were isolated by column chromatographies of silica ge(?)D101., polyamide,Sephadex LH-20,ODS and HPLC.On the basis of the chemical evidence and spectral analysis,the structures of 41 compounds were elucidated,they are 11,21-dioxo-3β,15α-dihydroxy-urs-12-en-2-O-β-D-glucoside(1),11,21-dioxo-3|3,15α-dihydroxy-urs-12-en-24-O-P-D-glucoside(2),11,21-dioxo-3β,15α-dihydroxy-olean-12-en-24-O-β-D-glucoside(3),apigenin-7-O-(2′-O-(5′′′-O-feruloyl)-p-D-apiofuranosyl)-β-D-glucopyranoside(4),chrysoeriol-7-O-[2′′-O-(5′′′-O-feruloyl)-β-D-apiofuranosyl]-β-D-glucopyranoside(5),(luteolin-7-O-[2′′-O-(5′′′-O-feruloyl)-β-D-apiofuranosyl]-p-D-glucopyranoside(6),(6-C-β-D-glucosyl-8-C-P-D-galactosyl-apigenin)(7),naringein-7-O-(2-O-β-D-apiofuran-osyl)-β-D-glucopyranoside(8),apiin(9),luteolin-7-O-β-D-glucopyranoside(10),chrysoeriol-7-O-β-D-glucopyranoside(11),apigenin-7-O-β-D-glucopyranoside(12),chrysoeriol(13),luteolin(14),quercetin(15),apigenin(16), (2S)-1-O-(7Z,10Z,13Z-hexadecatrienoyl)-3-O-β-galactopyanosyl-glycerol(17),(2S)-l-O-butanonyl-3-O-β-galactopyranosyl-glycerol(18),4β-O-glucopyranosyl-senkyunolide(19), sedanolide(20),3-oxo-megastigma-4,8-dien-7-O-β-D-glucoside(21),3-hydroxy-megastigma-5,8-dien-7-one(22),falcariondiol(23),(9z)-l,9-heptadecadiene-4,6-diyne-3,8,11-triol(24), oplopandiol(25),isofraxidin(26),5-methoxy psoralen(27),5,8-dimethoxyl psoralen(28), 3-O-caffeoyl-quinic acid(29),3-trans-coumaroylquinic acid(30),4-allyl-2-methoxyphenol (31),trans-ferulic acid(32),trans-cinnamic acid(33),p-hydroxyphenylethanol ferulate(34), 2-(3-methoxy-4-hydroxy)-propane-l,3-diol(35),lunularin(36),lunularic acid(37),,D-allitol (38),β-sitosterol(39),benzolic acid(40),succinic acid(41),respectively.Among the fourty-one compounds,three are triterpenes,thirteen are flavones,three are alcoholic glycosides,two are pathalides,two are megastigamanes,three are polyalkynes,nine are phenylpropanoids,three are phenol acids and four others.Among them,1,2,3,4,5,18,19 and 21 are new compounds,6,7,8,17,22,24,25,26,30,34,35,36,37 and 38 are isolated from apium genus for the first time.The above mentioned results indicated that triterpenes, alcoholic glycosides,megastigamanes and phenol acids were isolated for the first time from apium genus,which would provide chemical substances for elucidating the bioactive constituents of Apium graveolens.Bioactivity of three triterpenes(1-3),fourteen flavonoids(4-16,18)and one alcoholic glycoside(17)from Apium graveolens was tested in this thesis.These compounds were assayed against NO production on LPS activated macrophages by Griess method.Among them,4 flavones(13,14,15 and 16)displayed high inhibitory effects,but flavonoid glycosides and other kinds showed no correlated activity.The above 18 compounds were evaluated by 1,1-diphenyl-2-picrylhydrazyl(DPPH)free radical scavenging method.The results revealed only flavonoids with 3',4'-dihydroxy in B ring(6,10 and 13)displayed scavenging activity,and the others displayed no scavenging activity.MTT assay was used to test the cytotoxic activity of these 18 compounds against A549, Hela,and MCF-7 rumor cell lines.Among them,flavonoid aglycones and flavonoid glycosides displayed no cytotoxicity,but only flavonoid glycosides with the unit of trans-ferulic acid(4 and 5)showed cytotoxicity towards different cells.And other compounds displayed no significant cytotoxic activity.On the basis of chemical and bioactive studies,this thesis confirmed that flavonoids were the major bioactive compounds of fresh celery(Apiun graveolens).