Studing On The Coumarins For The Targets Of Xanthine Oxidase And Transient Receptor Potential Channel

Coumarins are a group of natural active compounds which are widely distributed in angiosperms.For example,compound esculetin showed Xanthine Oxidas（XO）inhibitory activity,while compound imperatorin displayed Transient Receptor Potential（TRP）channel modulation activity.In this dissertation,five coumarins-rich traditional chinese medicine（TCM）:the fruit of Cnidium monnieri（L.）Cuss.,the root of Angelica pubescens Maxim.f.biserrata Shan et Yuan,the root of Angelica dahurica（Fisch.ex Hoffm.）Benth.et Hook.f.,the fruit of Psoralea corylifolia L.and the aerial part of Leonurus japonicas Houtt.were selected as research targets.And the multi-target activities screening and material basis research were carried out.According to the inhibitory activity of XO and the modulation activity of TRP channels,the ethyl acetate extracts of C.monnieri and A.pubescens were choosed to research their activity components.Twelve compounds of C.monnieri extract were isolated and purified by high-speed counter-current chromatography（HSCCC）combined with high performance liquid chromatography（HPLC）,the structure of them were xanthotoxol（1）,merazin hydrate（2）, xanthotoxin（3）,isopimpinellin（4）,auraptenol（5）,bergapten（6）,cnidimol A（7）,osthenol（8）,imperatorin（9）,alloimperatorin（10）,osthol（11）and isomeranzin（12）.The effective extract of A.pubescens was subjected to separation by chromatographic column on Sephadex LH-20,ODS and HPLC,Their structures were identified as bergaptol（1）,isoimperatorin（2）,p-hydroxyphenylethanolferulate（5）,7-acetox-8-（3,3-dimethyl-oxiranylemthyl）-chromen-2-one（6）,demethylauraptenol（8）,7-demethylsuberosine（10）,psoralen（13）,decursidate（14）,caffeic acid methyl ester（15）,7-hydroxycoumarin（16）,5-hydroxy-6-prenylangelicin（17）,columbianadin（18）and six known coumarins.Among them,5-hydroxy-6-prenylangelicin（17）was a new compound,7-acetox-8-（3,3-dimethyl-oxiranylemthyl）-chromen-2-one（6）was isolated for the first time from natural sources,besides that the 1D and 2D NMR(¹H-NMR,¹³C-NMR and HMBC)spectral dates of 7-acetox-8-（3,3-dimethyl-oxiranylemthyl）-chromen-2-one（6）and demethylauraptenol（8）were first reported in this study.The inhibitory activity of XO and the modulation activity of TRP channels of compounds were screened.Xanthotoxol,xanthotoxin,bergaptol,demethylauraptenol,psoralen,7-hydroxycoumarin and 5-hydroxy-6-prenylangelicin exhibited a certain degree of XO inhibitory activity at the concentration of 200μM.The inhibitory rate of those compounds were 44.20±0.77%,43.67±1.44%,44.80±0.51%,47.30±0.88%,40.65±0.89%,54.74±0.76%and 81.39±0.17%,respectively.The modulation activity of TRP channels showed that osthol had inhibitory effects on TRPV1 and TRPV3 channels at the concentration of 200μM,the inhibitory rate were 40.20±5.77%and 54.07±5.11%,respectively.Xanthotoxol,cnidimol A,osthenol,alloimperatorin and columbianadin showed inhibitory effect on TRPV3 channels,the inhibitory rate of those compounds were 55.12±3.07%,50.54±3.39%,66.43±7.06%,50.59±11.12%and 64.22±6.06%,respectively.Based on the target activities of XO and TRP channels,the coumarins isolated from TCM were screened and studied,which provides a scientific basis for the research of gout drugs.