Studies On Synthesis Process,Quality Standard And Stability Of Class 1 New Antianginal Drug PF11

Pseudo-ginsenoside F11（PF11）is a characteristic ocotillol-type ginsenoside in American ginseng.Modern pharmacological experiments showed that PF11 had many pharmacological activities such as the the protective effects of the cardiovascular system,nervous system,lung and kidney.Considering that the content of PF11 in American ginseng is very low,it is difficult to realize industrial production.In order to realize the development and industrialization of PF11,a class 1 new antianginal drug,our group had converted the rich dammarane-type ginsenosides into octilone-type ginsenoside PF11 by means of structural modification.According to the scientific problems involved in the industrial production and development of PF11,such as the raw materials for the synthesis of PF11 and its dropping pills,synthesis process,quality standards and stability,this dissertation focused on the chemical component analysis of panax quinquefolium triolsaponins（PQTS）,the optimization of the synthesis process of PF11 and the related substances,the quality standards and stability of PF11 and its dropping pills.Based on the above studies,the quality standards of PF11 and its dropping pills were established,the stability of PF11 and its dropping pills were further studied,and the protective effect of PF11 on ischemic myocardium in vitro experiments were carried out.The results provided scientific data for the new drug application of PF11.1.The analysis of chemical components of PQTSFor the first time,the ultra performance liquid chromatography quadrupole-time of flight mass spectrometry（UPLC-Q/TOF-MS）technology combined with UNIFI platform was used to rapidly analyze and identify the chemical components of PQTS,which was the raw materials for the synthesis of PF11.A total of 31 triterpene saponins were identified,among which 10 had changes in the position of C-20 side chain and 21had no changes.2.Optimization of the synthesis process of PF11For the first time,single-factor investigation combined with Box-Behnken Design-response surface method was used to optimize the process of obtaining ginsenoside Rg2by alkaline hydrolysis of PQTS.The optimized parameters were as follows:the reaction alkalinity was 7.6%,the reaction temperature was 185℃and the reaction time was 5hours.The synthesis process of PF11 prepared by oxidative cyclization of Rg₂ was optimized by single-factor investigation experiment,and the optimized parameters were as follows:the reaction temperature was 75℃,the reaction time was 6 minutes and the oxidant dosage was 1.2 eq.3.Isolation and identification of the related substances of PF11Using some separation and purification methods such as macroporous resin,normal-phase and reversed-phase silica gel column chromatography,recrystallization,a total of six related substances of PF11 in the synthesis process were isolated.The structures of these compounds were identified by means of physical and chemical property,nuclear magnetic resonance（NMR）and high resolution mass spectrometry（HR-MS）.Among them there were four dammarane-type ginsenosides and two ocotillol-type ginsenosides.4.Establishment of the quality standards of PF11 and its dropping pillsFor the first time,according to the guiding principles and relevant regulations of"Technical Guiding Principles of the Standardization Process for the Establishment of Chemical Drug Quality Standards"and"Pharmacopoeia of the People’s Republic of China"（2020 version）,the properties,identification,inspection and content determination of PF11 and its dropping pills were studied.Then the quality standards（drafts）of PF11 and its dropping pills were established.5.Stability tests on PF11 and its dropping pillsFor the first time,according to the stability testing guidelines of active pharmaceutical ingredient（API）and preparation in the 9001 of Chinese Pharmacopoeia（2020 edition,VolumeⅣ）,the influence factor test,accelerated test and long-term test were carried out on PF11 and its dropping pills,in order to provide theoretical basis for the expiry date and storage conditions and ensure the safety.6.Study on the protective effect of PF11 on ischemic myocardiumFor the first time,the effect of PF11 on the hemodynamics of the isolated rat heart with myocardial ischemia reperfusion injury（MIRI）was studied.The Langendorff isolated heart perfusion system was used to establish the isolated rat heart MIRI model and the BL-420VHD biological function experimental system was used to detect the changes of isolated heart hemodynamic.The experimental results showed that PF11could inhibit the decrease of the LVDP,reduce LVEDP,alleviate left ventricular systolic and diastolic dysfunction in isolated rat hearts induced by MIRI,and enhance cardiac beating function.Among them,the high-dose administration group（50μg/m L）exerted the most obvious effect.In conclusion,this study provided pharmacy and pharmacodynamic data support for the further development of the active pharmaceutical ingredient PF11 and its dropping pills as an innovative class 1 chemical antiangina drug.