Studies On Chemical Constituents And Their Biological Activities Of Vitex Negundo Var.heterophylla

Vitex negundo L.var.heterophylla(Franch.)Rehd.,an aromatic shrub belonging to the family of Verbenaceae,is widely distributed in the hilly areas of the north,northeast,northwest,and central regions of China.All parts of this plant have been commonly used as folk medicine.Its leaves and branches are used to treat cold,fever,stroke,and urinary bleeding.Its roots and branches can be used for the therapy of bronchitis,malaria,and hepatitis.The essential oils from the seeds and leaves are utilized for the treatment of chronic bronchitis.By means of various chromatographic methods such as silica gel,Sephadex LH-20,opening ODS column chromatography,and semi-preparative HPLC,60 compounds were isolated from the 95%ethanol extract of the seeds and leaves of Vitex negundo var.heterophylla.Their structures were elucidated on the basis of spectroscopic data and physico-chemical methods.The compounds were comprised by 11 phenolic glycosides,10 phenylpropanoids,10 terpenoids,17 flavonoids,2 coumarins,and 10 others.Among them,compounds 1-4,13-15,22,and 52-55 were new,and named as vitexnegheteroins A-D(1-4),G(13),(3R,4S)-6,7-dihydroxy-4-(4-hydroxy-3-methoxyphenyl)-3-hydroxymethyl-3,4dihydro-2-naphthaldehyde-3?-O-?-D-glucopyranoside(14),vitexnegheteroins F(15),H(22),and I-L(52-55),respectively.The known compounds were identified as methyl(6-O-4-hydroxybenzoyl)-?-D-glucopyranoside(5),breynioside A(6)dunnianoside D(7),2-methyl pyromeconic acid 3-O-?-D-glucopyranoside-6'-(O-4"-hydroxybenzoate)(8),salviaplebeiaside(9),vitexfolin C(10),1,6-di-O-4-hydroxybenzoyl-?-D-glucopyranoside(11),vanilloyl-?-D-(2'-O-4-hydroxybenzoyl)-glucoside(12),vitexnegheteroin G(13),vitexnegheteroin E(14),vitexnegheteroin F(15),vitexdoin A(16),vitedoin A(17),6-hydroxy-4-(4-hydroxy-3-methoxyphenyl)-3-hydroxymethyl-7-methoxy-3,4-dihydro-2-naph thaldehyde(18),vitecannaside B(19),9-hydroxysesamin(20),3,5-di-O-caffeoyl quinic acid methyl ester(21),vitexnegheteroin H(22),2?-hydroxyursolic acid(23),arjunglucoside(24),negundoin G(25),vitexilactone(26),vitexilactone C(27),agnuside(28),5,4'-dihydroxy-3,6,7-trimethoxyflavone(29),5,4'-dihydroxy-3,6,7,8-tetramethoxyflavone(30),5,4'-dihydroxy-3,7,3'-trimethoxyflavone(31),5,4'-dihydroxy-6,7,3'-trimethoxyflavone(32),5,4'-dihydroxy-3,6,7,3'-tetramethoxyflavone(33),5,4'-dihydroxy-3,6,7,8,3'pentamethoxyflavone(34),5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone(35),luteolin-7-O?-D-glucopyranoside(36),diosmetin-7-O-?-D-glucopyranoside(37),orientin(38),isoorientin(39),isovitexin(40),isoorientin-6"-O-caffeate(41),3-C-glucopyranosylapigenin(42),vanillin(43),protocatechuic acid(44),isovanillic acid(45),frambinone(46),4-(3',4'-dihydroxyphenyl)butan-2-one(47),coniferaldehyde(48),p-hydroxybenzaldehyde(49),p-hydroxybenzoic acid(50),p-hydroxybenzoic acid methyl ester(51),4,5-di-O-caffeoyl quinic acid methyl ester(56),luteolin(57),quercetin(58),5,6,3',4'-tetrahydroxy3,7-dimethoxyflavone(59),and trans-caffeic acid,respectively.Among them,5 is a new natural compound and compounds 6,7,11,24,30,32,35,37,42,46,and 47 were obtained from the Vitex genus for the first time.The isolates were evaluated for their anti-tumor,anti-inflammatory,antioxidant,and antibacterial activities.The cytotoxic activities of the phenylpropanoids and terpenoids isolated were evaluated against HepG2,HCT116,and A2780 cell lines,using doxorubicin as the positive control.Compounds 17 and 18 exhibited moderate cytotoxic activities against HepG2 and HCT116,respectively,while compounds 23 and 25 showed moderate cytotoxic activities against all the selected cancer cell lines.The n-BuOH extract of the seeds,bark,and leaves of Vitex var.heterophylla showed better inhibition of the dimethylbenzene induced auricle oedema in mice.The isolated compounds were further tested for their inhibitory activities on LPS-induced NO production in murine microglial BV-2 cell line by the Griess reaction.All the evaluated isolates exhibited inhibitory effects on LPS-stimulated NO production.Among them,compound 25 showed significant inhibitory effect with an IC50 value of 0.71?M,which was comparable to the positive control SMT.The antioxidant effects of the phenolic glycosides and phenylpropanoids were assessed using the DPPH and ABTS radical scavenging assays.Significant differences were observed in the DPPH and ABTS radical scavenging assays.In the ABTS assay,most of the isolates showed significant antioxidant activity,with compounds 16-18,and 21 being the most potent,with IC50 values of 1.43,1.63,1.71 and 1.37?M,respectively.The petroleum ether extract of the seeds of Vitex negundo var.heterophylla showed different levels of inhibition to all the selected bacteria.This paper revealed the chemical constituents of Vitex negundo var.heterophylla and their bioactivity,which will provide a basis for further development and utilisation of this plant.