Anti-myocardial Ischemic Substances And In Vivo Process Analysis Of Erigeron Breviscapus Injection

Traditional Chinese medicine injections are widely used in clinic,but their safety and rationality have attracted much attention.The study of pharmacokinetics to reveal the process of drugs in vivo is of great significance in guiding clinical safe and rational drug use.Due to the complexity of traditional Chinese medicine injections,the effective substances of traditional Chinese medicines are not yet clear,and whether the pharmacokinetics of single or several ingredients can characterize the whole in vivo process of traditional Chinese medicines need to be discussed and demonstrated.The Erigeron breviscapus injection is a sterile aqueous solution extracted by the whole dry plant of Erigeron breviscapus（Vant.）Hand.-Mazz.It is mainly used for cardiovascular and cerebrovascular diseases in clinic such as coronary heart disease,angina pectoris and acute cerebral infarction ect.At present,the effective substances with anti-myocardial ischemic effect of Erigeron breviscapus injection are unclear and the in vivo process remains to be studied further.This article took Erigeron breviscapus injection as the research object to study the in vivo anti-myocardial ischemia activity of its main components based on its material basis,to find out the main effective components,and to further explore multi ingredients pharmacokinetics study method using the major effective components as indicators.Through the above studies,we hope to reveal the effective components and the overall characteristics of Erigeron breviscapus injection in rats and to provide a new methodological reference for the pharmacokinetic study of multi ingredients in traditional Chinese medicine.The specific research contents and results are as follows:1.LC-ESI-MS/MS was used to deduce the fragmentations of the chemical components in Erigeron breviscapus injection,and combined with the accurate molecular weight information determined by LC-Q-TOF-MS,fifteen major components were identified.2.The quantitative analysis method of 13 components in Erigeron breviscapus injection by LC-ESI-MS/MS was established for the first time and it was used for the determination of 13 components in Erigeron breviscapus injection.The column was Ultimate^?XB-C₁₈（4.6×100 mm,3.5μm）,with a flow rate of 0.7 m L/min.The MRM was tested in the negative ion mode and all 13 compounds were detected within 24 min.The method was simple and rapid,and it was used for the determination of five batches of Erigeron breviscapus injection.The results showed that scutellarin is the highest in each batch（410.93±61.74μg/m L）;and the content of 4,5-dicaffeoylquinic acid and apigenin-7-O-glucuronide were the second highest（225.70±71.15μg/m L,157.97±67.18μg/m L）.The above studies provided content information for later studies.3.The mice model of myocardial ischemia induced by isoproterenol was established to evaluate the anti-myocardial ischemia efficacy in vivo of 10 major components in Erigeron breviscapus injection.10 major components are 5-caffeoylquinic acid,4-caffeoylquinic acid,3-caffeoylquinic acid,Caffeic acid,1,3-dicaffeoylquinic acid,Scutellarin,3,4-dicaffeoylquinicacid,3,5-dicaffeoylquinicacid,apigenin-7-O-glucuronide and 4,5-dicaffeoylquinic acid.The results showed that Scutellarin,3-caffeoylquinicacid,apigenin-7-O-glucuronideand4,5-dicaffeoylquinic acid can significantly reduce the ST-segment elevation of ECG in model mice and can significantly reduce the serum CK levels of model mice（vs.model group,P<0.05）,and can reduce the pathological damage of the myocardial tissue in model mice.The above studies showed that scutellarin,3-caffeoylquinic acid,apigenin-7-O-glucuronide,and 4,5-dicaffeoylquinic acid may be the main effective components of Erigeron breviscapus injection against myocardial ischemia.4.The quantitative analysis method of 13 components of Erigeron breviscapus injection in rat plasma by LC-ESI-MS/MS was established for the first time,and it was applied to pharmacokinetic study of Erigeron breviscapus injection in rats.The plasma concentrations of 13 components were detected and the pharmacokinetic parameters were calculated after intravenous administration of a low（7.2 m L/kg,10times volume concentrated before administration）and high dose（36 m L/kg,10times volume concentrated before administration）of Erigeron breviscapus injection.The results showed that the difference in the t_1/2of 13 components at two doses was not significant,but the AUC increased with the increase of dose.Scutellarin,Erigoster B,3,4-di CDOA or 4,9-di CDOA are relatively slowly eliminated in rats(t_1/2>1 h),and the other 10 components are rapidly eliminated in rats(t_1/2<1 h).The AUC of 4-CDOA,scutellarin,3,4-dicaffeoylquinic acid,3,5-dicaffeoylquinic acid,Erigoster B,3,4-di CDOA（or 4,9-di CDOA）,apigenin-7-O-glucuronide and4,5-dicaffeoylquinic acid the 8 components is relatively large(the low dose AUC_0-∞is 215.61-2275.72 h·ng/m L,and the high dose AUC_0-∞is 1422.05-7484.27 h·ng/m L).Considering the content,efficacy and pharmacokinetics of each component,scutellarin,3-caffeoylquinic acid,Erigoster B,3,4-di CDOA or 4,9-di CDOA,apigenin-7-O-glucuronide and 4,5-dicaffeoylquinic acid were selected as pharmacokinetic markers to characterize the pharmacokinetic behavior of Erigeron breviscapus injection in vivo;In addition,the pharmacokinetic markers were integrated using the plasma concentration sum method and AUC weight integration method.