| Vitex negundo var.cannabifolia,family Verbenaceae,is mainly distributed in the Yangzi River basin of China.The fruits possess the actions of removing moisture and sputum,relieving cough and asthma,and regulating qi to alleviate pain.And they are used in traditional Chinese medicine for the treatment of various diseases,such as cough and asthma,stomachache,diarrhea,colic pain,beriberi,leucorrhea,and gonorrhea.Our previous study has demonstrated that the CH2Cl2 extract of the fruits of V.negundo var.cannabifolia showed inhibition of nitric oxide(NO)production in lipopolysaccharide(LPS)-activated RAW 264.7 cells.In order to clarify the effective material basis and action mechanism,a bioassay-guided fractionation was performed on the chemical constituents of this plant.The inhibitory effect of the isolated compounds on NO production in LPS-stimulated RAW 264.7 cells and cytotoxic effect on human hepatocellular carcinoma cell line(HepG-2)were evaluated in vitro.In addition,pharmacokinetics,tissue distribution,and metabolism of the major constituents were studied in rats.The results are listed as below.1.The research progress of chemical constituents and bioactivities of the plants of V.negundo var.cannabifolia have been reviewed.The main chemical constituents of the plants from this species are lignans,flavonoids,diterpenoids,triterpenoids,irioids,aromatic acids and some other constituents.Crude extracts and pure compounds from Vitex plants have been found to exhibit a wide array of biological activities including antiasthmatic,analgesia,antibacterial,anti-inflammatory,antioxidant,and anti-tumor effects.2.The chemical constituents were separated and purified via various column chromatography including silica,sephadex LH-20,ODS,and semipreparative HPLC.As a result,twenty-nine compounds were isolated from the CH2Cl2 extract of the fruits,including fifteen lignans:6-hydroxy-4-(4-hydroxy-3-methoxyphenyl)-3-hydroxymethyl-7-methoxy-3,4-dihydro-2-naphthaldehyde(1),vitedoin A(2),vitexdoin F(3),detetrahydroconidendrin(4),vitexdoin D(5),vitexdoin E(6),4-oxosesamin(7),L-sesamin(8),paulownin(9),(+)-beechenol(10),ligballinol(11),2-(4-hydroxyphenyl)-6-(3-methoxy-4-hydroxyphenyl)-3,7-dioxabicyclo[3.3.0]octane(12),(-)-pinoresinol(13),balanophonin(14),thero-guaiacylglycerol-β-coniferyl aldehyde ether(15);fourteen aromatic acids:methyl p-hydroxy benzonate(16),p-ethylbenzoic acid(17),vanillin(18),trans-p-coumaryl aldehyde(19),coniferyl aldehyde(20),p-hydroxy benzaldehyde(21),p-hydroxybenzoic acid(22),ferulic acid(23),syringaldehyde(24),4-hydroxybenzeneethanol(25),sinapaldehyde(26),5,7-dihydroxychromone(27),frambinone(28),and alternariol 4-methyl ether(29).Among them,Compounds 10-12,19,28,and 29 were firstly isolated from Verbenaceae family,compound 15 was firstly obtained from the genus Vitex,and compounds 7,8,14,20,26,and 27 from this plant for the first time.3.The NO inhibitory effect of the isolated compounds on LPS-stimulated RAW 264.7 cells and cytotoxic effect on human hepatocellular carcinoma cells(HepG-2)were evaluated in vitro.As a result,compounds 1,3-8,11-13,18-20,23,and 26 showed significant inhibition of nitric oxide production in LPS-stimulated RAW 264.7 cells,compounds 1-4 showed cytotoxicity on HepG-2 cells.4.After intragastric administration of CH2Cl2 extract,pharmacokinetic profiles and tissue distribution of three compounds(compounds 1,2,and 4),which possess activities such as anti-inflammatory,anti-tumor,and antioxidant activities,were studied in rats.The concentration-time curves and pharmacokinetic parameters were obtained for these three compounds,which could help to clarify the effective material basis and action mechanism of the fruits of V.negundo var.cannab,ifolia.5.Metabolites in rat feces,urine,bile,and plasma were characterized following the oral administration of two major compounds(compounds 1 and 2),two protypes and eleven metabolites were plausibly annoted.Among the metabolism,glucuronidation,protype,and hydroxylation metabolic pathways were observed in rat plasma,glucuronidation and hydroxylation in rat bile,glucuronidation and protype in rat urine,and protype in rat feces. |
