Study On The Formation Mechanism Of Isopropyl Ester Metabolites By ’Liang Guanxi’ Herb Pairs Salvia Miltiorrhiza-Dalbergia Odorifera

Ischemic stroke is a common and high-incidence type of cardiovascular and cerebrovascular diseases,which seriously threatens human healthy life.The method of activating blood and removing blood stasis is a common clinical treatment method,and Salvia miltiorrhiza and Dalbergia odorifera belonged to the category of typical medicine pairs for activating blood and removing blood stasis.Preliminary research of the research group on the in vivo effect substances of the "Liang Guanxi" Salvia miltiorrhiza and Dalbergia odorifera herb pairs,it was found that compared with the single use of Salvia miltiorrhiza,the combination of the two drugs can not only enhance the anti-cerebral ischemia activity,but also identify the core effect substance Isopropyl3-(3,4-dihydroxyphenyl)-2-hydroxypropanoate(IDHP)and Isopropyl(E)-3-(3,4-dihydroxyphenyl)acrylate(KYB).Thus,we suspected that the drug phenolic acid compounds and endogenous ketogenic amino acids are connected to the formation of two esters.This article conducts a preliminary study on the formation mechanism of the above two esters through the following content:1.Established the UPLC-MS/MS analysis method of IDHP and KYB in rat tissue samples,established cerebral ischemia injury rat as an animal model,and investigated the administration of Danshen injection and Xiangdan injection The similarities and differences of the tissue distribution of the two esters after the drug.The results showed that only KYB was produced in the tissues of Danshen injection group,while both kinds of esters were produced in the tissues of Xiangdan injection group,and the tissues with more distribution of both esters were heart,brain and spleen.Compared with Danshen injection,the production of KYB is significantly increased,and its distribution trend changes from kidney,brain,and liver to heart,brain,and kidney.The results were consistent with the previous pharmacodynamics results,which provided a basis for the "Liang Guanxi" between Salvia miltiorrhiza and Dalbergia odorifera,and laid a foundation for the study on the formation mechanism of isopropyl ester metabolites in the later period.2.Established the UPLC-MS/MS analysis methods of Danshensu,caffeic acid,salvianolic acid A,protocatechuic acid,rosmarinic acid,protocatechualdehyde;tryptophan,leucine,phenylalanine,lysine,tyrosine,isoleucine,threonine,etc.,use the heart,brain,and spleen tissues with high production of two esters to analyze the relationship between the above two esters with salvianolic acids and endogenous ketogenic amino acids by means of SPSS.From the results of correlation analysis,it is proved that the formation of IDHP is strongly related to the pharmaceutical ingredients Danshensu,Salvianolic Acid A,and endogenous tryptophan,tyrosine,leucine,and isoleucine;caffeic acid The formation of KYB is strongly causally related to the drug components rosmarinic acid,protocatechualdehyde,and endogenous tyrosine.3.In view of the structural characteristics of IDHP and KYB,and in combinationed with the results of the third chapter,two key links in the formation of esters,namely,danshensu/caffeic acid to produce isopropyl danshensu/isopropyl caffeate as a key point,in vitro bio-simulation technology was used to verify the generation of IDHP / KYB and explore the probable influencing factors.The results showed that isopropanol and Danshensu/Caffeic acid can generate IDHP / KYB in vitro,and Coenzyme A is one of the effective catalytic enzymes.