Study On Material Basis Of 'liang Relationship' Herb Pair Salviae Miltiorrhizae Radix Et Rhizoma-Lignum Dalbergiae Odoriferae In Vivo

It is an urgent task for pharmaceutical researchers to explore safe and effective drug for the prevention and treatment of cardiovascular and cerebrovascular diseases which remains the number one killer that endangers human health.Chinese herbal compound is the source of developing highly effective innovative drug,thus to clarify the pharmacodynamic material basis is both the key link and bottleneck.Herb pair,the core part of Chinese herbal compound,reflects the vital compatibility relationship among the drugs.From the perspective of herb pair,it is an important breakthrough point to study the material basis of Chinese herbal compound.Under the guidance of identification thought of effective components cluster for ?Liang relationship‘ herb pair previously,Salviae miltiorrhizae Radix et Rhizoma-Lignum Dalbergiae Odoriferae(SM-LDO),the classical herb pair for invigorating blood circulation and eliminating stasis,was selected as the object of the research and Xiangdan Injection(XDI)was set as the carrier of the research,owing to its stable and controllable quality.The pharmacodynamic material basis of SM-LDO herb pair in the normal and acute blood stasis model rats was systemically studied from the point of pharmacokinetics.The purpose of this present thesis is to provide the basis for the research of the material basis on Chinese herbal compound in the future.The dissertation was divided into 3 chapters.The main contributions of the author were as follows:1.The analytical methods of HPLC-Q-TOF/MS and GC/MS for XDI were developed.The basic substances of XDI were characterized and identified,which contained the 53 water-soluble components and 9 volatile components.The results above could provide the traceability of basic substances for the establishement of quality control and quality standard of SM-LDO,and also lay methodology foundation for the metabolism of SM-LDO herb-pair in vivo under the same analytical conditions.2.An HPLC analytical method for salvianolic acids in rat plasma was established.The established HPLC analytical method and the above HPLC-Q-TOF/MS analytical method were applied to the metabolic profile analysis in vivo for the SM-LDO herb-pair in normal and acute blood stasis rats.A total of 39 drug prototype components(mainly phenolic acids)and 79 drug metabolites were identified,and the major metabolic pathways involved with methylation,sulfation and glucuronidation.The differences of the drug metabolism of SM-LDO herb-pair in normal and acute blood stasis rats were compared.The results showed that pathological factors could increase the content of drug into the blood,slow down the elimination of drug,and prolong the effective time of drug,compared with normal rats.3.The tissue distribution(heart,brain,liver and kidney)of SM-LDO herb-pair in normal and acute blood stasis rats were conducted by the proposed HPLC-Q-TOF/MS analytical method.The total of 24 prototype and 76 metabolites of the herb-pair were identified,which were mainly distributed in the heart,liver and kidney.Specially,salvianolic acids such as Danshensu,rosmarinic acid,salvianolic acid A were mainly absorbed into the target organs with rich blood flow like heart,brain and liver,and the main metabolic forms involved with methylation,glucuronidation and sulfation in vivo.The isopropylated metabolic form and related products were found and confirmed in many metabolic forms of acute blood stasis rat tissues.IDHP was confirmed in rats and isopropyl caffeate was firstly discovered.The study indicated that the isopropylated metabolic form might be an important and characteristic metabolic pathway of ?formula corresponding to syndrome‘ on ?Liang relationship‘ herb pair SM-LDO in rats.4.Using principal component analysis,the correlation of the significantly changing ingredients(drug components,endogenous substances and metabolites)in dosed plasma of rat was analysed by means of SPSS statistical software.Combined with the research results of the tissue distribution above,the material basis of SM-LDO herb pair in vivo was preliminarily confirmed,including rosmarinic acid,salvianolic acid A,IDHP,isopropyl caffeate,methylsalvianolic acid A glucuronide,dimethylsalvianolic acid A and dimethylsalvianolic acid A glucuronide.5.The metabolic process of IDHP,one of material basis of SM-LDO,was preliminarily investigated.The main metabolites of IDHP in vivo were Danshensu and its metabolites.The results of drug metabolism provided strong support for better understanding of the efficacy and safety of IDHP.An analytical method of HPLC-Q-TOF/MS for the detection of Danshensu in human urine was also established and used for the structure confirmation of Danshensu in the urine of patients under anoxic and inflammatory conditions.(see Appendix).