Studies On Active Components Of Xuesaitong Injection And Their In Vivo Processes

Currently, remarkable achievements have been obtained in the field of Chinese medicine basis investigation in China. Due to the application of modern scientific theory and technology in modernizaiton innovation of Traditional Chinese Medicine (TCM), the modern TCM industries have been enormously promoted and developed into strategic emerging ones. To further raise the scientific competitive potential, impel the internationalization process of TCM industries, and enroll TCM in the main stream international medicine market, it is necessary to further investigate the chemical material basis, active components and effect mechanisms, and multiple components’ pharmacokinetics of TCM by advanced modern technological methods to disclose the scientific connotation of TCM and guide scientifically the usage of TCM products in clinic.Xuesaitong injection, a preparation of total saponin extracts of Panax notoginseng with significant medical efficancy, which has been widely used in clinic, is a big species of TCMs proessing tremendous development perspective. Therefore, in this dissertation, with Xuesaitong injection as a carrier, some key and common scientific problems about the further development of big TCM species, involving chemical components, methodology of drug quality analysis, therapeutic material basis, in vivo processes of main components, etc. for Chinese patent medicine, were investigated by using the comprehensive techniques and methods of mutli-crossed disciplines, which especially focused on some difficult points, such as multicomponent pharmacokinetics and PK-PD model studies, etc.The main contents and academic contributions of this thesis are summarized as follows:1 The chemical components of Xuesaitong injection were studied using HPLC-ESI-MS". As a result,27 saponins were identified in the preparation.2 With only 10 saponins references, an HPLC method for simultaneous determination of 27 saponins in Xuesaitong injection was developed. In addition, a method for HPLC fingerprinting analysis of Xuesaitong injection was also established. Finally, quality control method for Xuesaitong injection was developed by combination of the developed quantitative and fingerprinting analysis methods.3 A high through-put screening platform for antioxidative active components from TCMs was developed based on H2O2-Luminol and DPPH·-Luminol chemiluminesce systems and the platform was successfully used to evaluate the antioxidative capacity of the main chemical components of Xuesaitong injection. Further, the antioxidation and antihypoxia effect of the injection and its main components were also evaluated by H9C2 cell model in vitro. The results suggest that the 12 main saponins in Xuesaitong injection have antioxidative effects in vitro. In addition, Rg1 and Rb1 had obvious protection effect against H9C2 cell injury caused by hypoxia.4 Using the rat model of myocardial ischemia, the therapeutic effect of Rb1 and Rgi (the two main chemicals of Xuesaitong injection) were evaluated. The results demonstrated that Rg1 had significant therapeutic effect on myocardial infarction rat models, which suggested that Rg1 was the main active component of Xuesaitong injection.5 An HPLC-TQMS method was developed for simultaneous determination of 12 main saponins (R1, Rg1, Re, Rg2, Rh1, Rc, Rb1, Rb2, Rb3, Rd,20(S)-Rg3 and 20(R)-Rg3) from Xuesationg injection in rat plasma and tissues. With the developed method, the in vivo processes of the 12 saponins above-mentioned were investigated in rats.The results demonstrated that the main active components of Xuesaitong injection were predominantly distributed into kindney, lung, muscle and heart, suggesting that these tissues and organs were the main action or accumulation positions of active components; pharmacokinetic profiles of the 12 saponins could be divided into fast elimination type (including R1, Rg1, Re, Rg2, Rh1, SRg3 and RRg3) and slow elimination type (including Rc, Rb1, Rb2, Rb3 and Rd). Considering that both Rg1 and Rb1 are the main chemicals of Xuesaitong injection, possess the pharmacokinetic property of fast elimination and slow elimination, respectively, and can represent the two kinds of saponins in Xuesaitong injection, respectively, we proposed originally that Rg1 and Rb1 were selected as pharmacokinetic markers (PK markers) for pharmacokinetic evaluation of Xuesationg injection.6 The characteristic metabolic biomarkers for myocardial infarction were found by metabolomic investigation on myocardial infarction rat models. Based on the discoveries, metabolomic evaluation methodology for therapeutic effect of TCM was developed. Finally, according to the research idea above mentioned, the PD-PK link investigation for Xuesationg injection was originally proposed based on metabolomic technique. According to the proposed idea, with Rg1 as PK markers and the characteristic metabolic biomarkers for myocardial infarction rats as pharmacodynamic markers, the PD-PK link investigation for Xuesationg injection was performed. The results domenstrated that a PK-PD link model was well fit for Rg1 and lactic acid and it could predict the in vivo effectiveness change of Xuesaitong injection by monitoring the concentration of Rg1 in plasma, which provides a reference and evidence for designing the administration scheme of Xuesaitong injection in clinic. In addition, the results above mentioned also suggest that Rg1 could exert the therapeutic effect aginst myocardial infarction through the glycolysis pathway. The presented study could also provide a novel methodological reference for TCMs’PD-PK link investigation.