Metabolites Of Caffeoylquinic Acid In Dengzhanhua And Their Anti-stroke Related Effect

Erigeron breviscapus is one of the most important national medicines in Yunnan.It has the effects of activating blood circulation,dredging collaterals and relieving pain,dispelling wind and dispelling cold,and the minority nationalities of Han nationality,Miao nationality,Zhuang nationality,Bai nationality,and so on.The content of caffeic acid quinic acid compounds in Erigeron breviscapus is high,and pharmacological studies show that these compounds have antioxidant,anti-free radical,anticoagulant,anti-fibrosis and other effects,can play a role in different stages of ischemic stroke The But the bio-availability of caffeic compounds is low.In this study,caffeoyl compounds as the object of study,proposed with the parental caffeic acid metabolites may exist in the "multi-source homing" characteristics,common metabolites with anti-stroke activity.Objective:In this paper,HPLC method was used to determine the content of caffeine compounds in Erigeron breviscapus and its preparations.Blood and urine samples were collected by oral administration.The common metabolites were searched and the common metabolites were identified by UPLC-QTOF MS.The antioxidant activity of the metabolites and the prototype products was evaluated by the DPPH/ABTS.The coagulation activity of the caffeic acid group and its metabolites was compared with four samples.The levels of IL-6,TGF-? and NF-?B were used to compare the caffeic acid And its metabolic components anti-inflammatory activity,the evaluation of nerve cell protection,analysis of caffeic compounds metabolites,prototypes and metabolites of the pharmacological activity of comparative study of the comparative study.Result:1,Erigeron breviscapus extract is mainly wort baicalein,lamp brassinide,scutellarin and other flavonoids(peak area of about 25%)and mono-caffeoylquinic acid,co-caffeic acid and the like(peak area of about 60%);2,This compounds of caffeic acid(CA),3-caffeoylquinic(3-CQA),4-caffeoylquinic(4-CQA),5-caffeoylquinic(5-CQA),1,3-dicaffeoylquinic acid(1,3-DCQA),3,5-dicaffeoylquinic(3,5-DCQA),4,5-dicaffeoylquinic(4,5-DCQA),was administered and the urine was collected,The common metabolites were identified as Ferulic acid(FA),Isoferulic acid(IFA),Dihydro ferulic acid(DHFA),m-hydroxybenzenepropanoic acid,dihydrogen coffee Acid and so on;3,in the urine,blood,brain,stomach,feces and other biological samples found that caffeoyl compounds are a lot of metabolism,caffeic compounds in the body mainly caffeic acid,ferulic acid and other metabolites,the prototype product content is too low Not detected.4,The concentration of 0.05mg/mL?0.55mg/mL of the prototype/metabolic compounds for DPPH/ABTS antioxidant activity test results show that the common metabolite metabolite FA only remove the ABTS free radical activity is good,the prototype product bacryloyl Preferably,The DPPH free radical scavenging capacity(IC50 value)was:FA(0.725mM)<3-CQA(0.227mM)<4-CQA(0.196mM)<5-CQA(0.0.193mM)<VC(0.131mM)<3,5-DCQA(0.118mM)<4,5-DCQA(0.099mM)<CA(0.098mM)<3,4-DCQA(0.096mM)<1,3-DCQA(0.088mM);The ABTS free radical scavenging capacity(IC50 value)was:VC(0.298mM)<4-CQA(0.192mM)<5-CQA(0.19mM)<3-CQA(0.188mM)<3,5-DCQA(0.154 mM)<3,4-DCQA(0.134mM)<CA(0.098mM)<4,5-DCQA(0.091mM)<1,3-DCQA(0.082mM)<FA(0.053mM).On-line HPLC-ABTS results show that both metabolites and prototypes have antioxidant activity,and many metabolites in the urine are highly resistant to oxidation,but the structure is unknown.5,the total metabolites had a good anticoagulant activity at 3 mg/mL,compared with the blank control group(11.4 s),P<0.01,the prothrombin time(PT)was significantly prolonged:CA((11.85 s),FA(12.85 s),IFA(12.85 s),DHCA(11.75 s)and DHFA(12.7 s)were significantly longer than those in the blank group(38.37 s)(73.3 s),FA(108.65 s),IFA(108.85 s),DHCA(75.35 s)and DHFA(100.7 s),respectively.Compared with the blank group(27.43 s),the activated partial thromboplastin time(APTT)(30.15 s),DHFA(30.15 s);the activity of the prototype product at the same concentration was not significant,and the concentration of 1.5 mg/mL was 1.5 mg/mL Under the prototype and metabolite activity were not obvious.6,compare caffeic acid prototype compounds and their metabolites anti-inflammatory activity,the results show that the expression of IL-6 factor IC50 value size:3-CQA(347.1?M)>DHFA(336.1?M)>DHCA(141.2?M)>CA(120.8?)>IFA(101.8?M)>FA(69.74?M)>4-CQA(53.63?M)>3,5-DCQA(34.83?M)>3,4-DCQA(33.35?M)>4,5-DCQA(32.6?M)>1,3-DCQA(30.48?M),metabolites activity is low,double caffeoyl quinine acid activity;affect the expression of TGF-? factor IC50 value size:CA(340.3?M)>FA(299.4?M)>IFA(248.7?M)>(210.0?M)>3,5-DCQA(117.47?M)>3,4-DCQA(107.29?M)>1,3-DCQA(102.2?M)>DHCA(92.59?M)>4-CQA(66.57?M)>4,5-DCQA(64.2?M)>3-CQA(63.68?M),The activity of metabolites is low,the prototypic caffeyl quinine acid activity is high;affect the expression of NF-?B factor IC50 value size:IFA(624.2?M)>FA(592.2 ?M)>CA(450.0?M)>1,3-DCQA(364.2?M)>3,4-DCQA(332.9?M)>4,5-DCQA(185.3?M)>3,5-DCQA(140.4?M);metabolites activity is low,double caffeoyl quinic acid activity is high.Conclusion:Caffeoyl compounds have many properties of homologous metabolites,and caffeoyl compounds are metabolized mainly in caffeic acid,ferulic acid and isoferuric acid,and no prototypes are found.Compounds,and the common metabolites in the antioxidant,anti-inflammatory,anticoagulant activity were weak,and the activity was less than the prototype product of briacenoyl quinine acid.Experiments have shown that the common metabolites of the caffeic compounds are weakly active in the study of stroke in the treatment of Erigeron breviscapus,and the prototype product is not detected in the body.Therefore,whether the caffeic compounds need to be combined Dynamics and its organization distribution for further quantitative evaluation.