Study On The Anti-inflammatory Constituents Of Canarium Album (Lour.) Rauesch.

Qingguo,also known as Chinese olive,green olive,and white olive,is the dried ripe fruit of Canarium album(Lour.)Rauesch.,a genus of Canarium in the Burseraceae family.As a traditional food and medicine,Qingguo is a common Chinese medicine for clinical treatment of sore throat and thirst,as well as a preserved fruit for young and old in the south of China.Pharmacological research shows that Qingguo has many biological effects,such as anti-inflammatory,anti-virus,hypoglycemic and hypolipidemic,and anti-tumor.In this thesis,we investigated the anti-inflammatory components of the chloroform and ethyl acetate parts of 70%ethanol extract of Qingguo.The separation and purification were carried out using silica gel column chromatography,Sephadex LH-20 column chromatography,ODS MPLC,and preparative HPLC.Finally,seventy-two compounds were isolated and identified:(1)Benzofuran neolignans:(+)-erythro-(7S,8R)-canariol A(1a*),(-)-erythro-(7R,8S)-canariol A(1a*),(+)-threo-(7R,8R)-canariol A(2a*),(-)-threo-(7S,8S)-canariol A(2b*),(+)-erythro-(7S,8R)-canariol B(3a*),(-)-erythro-(7R,8S)-canariol B(3b*),(+)-threo-(7R,8R)-canariol B(4a*),(-)-threo-(7S,8S)-canariol B(4b*),(+)-(7S,8R,8’R)-canariol C(5a*),(-)-(7R,8S,8’S)-canariol C(5b*),(+)-(7S,8R,8’S)-canariol C(6a*),(-)(7R,8S,8’R)-canariol C(6b*),(+)-(7S,8R,7’S,8’S)-hawthornnin E(7a*),(-)-(7R,8S,7’R,8’R)-hawthornnin E(7b*),(+)-(7S,8R,7’R,8’R)hawthornnin E(8a*),(-)-(7R,8S,7’S,8’S)-hawthornnin E(8b*),(+)(7R,8R,7’S,8’R)-picrasmalignan(9a*),(-)-(7S,8S,7’R,8’S)-picrasmalignan(9b*),(+)-(7R,8S,7’S,8’R)-picrasmalignan(10a*),(-)-(7S,8R,7’R,8’S)picrasmalignan(10b*),(+)-(7S,8R,7’S,8’R)-picrasmalignan(11a*),(-)(7R,8S,7’R,8’S)-picrasmalignan(11b*),(+)-(7S,8R,7’S,8’S)-picrasmalignan(12a*),(-)-(7R,8S,7’R,8’R)-picrasmalignan(12b*),(+)-(7S,8R,7’S,8’R)cinncassin D(13a*),(-)-(7R,8S,7’R,8’S)-cinncassin D(13b*),(-)(7S,8R,7’R,8’S)-cinncassin D(14a*),(-)-(7R,8S,7’S,8’R)-cinncassin D(14b*),dihydrodehydrodiconiferyl alcohol(15),Prunustosanan AI(16).(2)Phenylpropanoids:(-)-(7R,7’R,7"R,8S,8’S,8"S)-canariol D(17a*),(+)(7S,7’S,7"S,8R,8R,8"R)-canariol D(17b*),(+)-(7R,7’R,7"S,8S,8’S,8"R)canariol D(18a*),(-)-(7S,7’S,7"R,8R,8’R,8"S)-canariol D(18b*),(-)(7R,7’R,7"R,8S,8’S,8"R)-canariol D(19a*),(+)-(7S,7’S,7"S,8R,8’R,8"S)canariol D(19b*),(-)-(7R,7’R,7"S,8S,8’S,8"S)-canariol D(20a*),(+)(7S,7’S,7"R,8R,8’R,8"R)-canariol D(20b*),salicifoliol(21),pinoresinol(22),epipinoresinol(23),crataegusnin E(24),crataegusnin F(25),threo2,3-bis-(4-hydroxy-3-methoxyphenyl)-3-methoxy-propanol(26),erythro2,3-bis-(4-hydroxy-3-methoxyphenyl)-3-methoxy-propanol(27),threocrataegusal A(28),threo-guaiacyl-glycerol-β-O-4’-dihydroconiferyl ether(29),threo-guaiacylglycerol 8-vanillin ether(30),threo-guaiacylglycerolβ-coniferyl aldehyde ether(31),erythro-guaiacylglycerol 8-vanillin ether(32),erythro-guaiacylglycerol-β-coniferyl aldehyde ether(33),7hydroxyl-dihydroferulic acid ethyl ester(34),3-(4’-hydroxy-3’methoxyphenyl)-1-propanol(35),p-hydroxy cinnamaldehyde(36),methyl p-hydroxycinnamate(37).(3)Phenolic glycosides:1-ethyl-6-cinnamoylβ-D-glucopyranoside(38),1-methyl-6-cinnamoyl-β-D-glucopyranoside(39),3,4-dimethoxyphenyl-(6’-O-4"-hydroxy benzoyl)-O-β-Dglucopyranoside(40),3,4-dimethoxyphenyl-(6’-O-vanilloyl)-O-β-Dglucopyranoside(41).(4)Phenols:4,4’-dihydroxy-3,3’-dimethoxy benzophenone(42),vanillil(43),4-methoxybenzene-1,2-diol(44),phydroxybenzaldehyde(45),4-hydroxyacetophenone(46),gallic acid(47).(5)Flavonoids:apigenin-7-O-β-D-glucopyranoside(48),chrysoeriol-7-Oβ-D-glucoside(49),apigenin-7-O-(3"-E-p-coumaroyl)-glucopyranoside(50).(6)Others:1H-indole-3-carboxaldehyde(51),cyclo-(Phe-Leu)(52),cyclo-(Phe-Val)(53),4-carbomethoxy-6-hydroxy-2-quinolone(54).Among the 72 identified compounds,38 were new compounds(1a/1b14a/14b,17a/17b-20a/20b,40,41).Based on the chemical composition studies,this thesis also investigated the in vitro anti-inflammatory activity of the main compounds.The results showed that all of the main compounds isolated from Qingguo showed significant inhibition of LPS-induced nitric oxide(NO)production in RAW 264.7 cells,with compounds 1b-3b,6a,9a-11a,11b,12a,15,16,18a,19b,28,31,33,36,38,41,48 and 50 had significantly lower IC50 values than the positive drug indomethacin.In addition,there were significant differences in IC50 values between the enantiomeric isomers of compounds 1a/1b,3a/3b,5a/5b,7a/7b,12a/12b,18a/18b,19a/19b and 20a/20b.Further studies revealed that compounds 5a,5b,6a,6b,9a,9b,20a,20b,40,and 41 could dose-dependently inhibit the expression of COX-2 and iNOS at the protein and mRNA levels,as well as the mRNA expression of the pro-inflammatory cytokines IL-1β and IL-6 in LPSstimulated RAW 264.7 cells.Subsequently,further studies on antiinflammatory mechanisms showed that compounds 5a,5b,6a,6b,9a,and 9b significantly inhibited the nuclear translocation of NF-κB in a concentration-dependent manner,which suggested that they exerted antiinflammatory effects by blocking the nuclear translocation of NF-κB,and thereby reducing the expression of pro-inflammatory mediators COX-2,iNOS,IL-1β,and IL-6.