Research On Salvia Miltiorrhiza Injection Of CYP450 Enzyme Inhibitory Effect Research And Identification Of Structure Of Honokiol Metablites In Rat Small Intestine

Drug discovery and development of drug metabolism is an integral part of research into the metabolism in vivo and in vitro metabolism studies. By exploring the metabolic characteristics in vitro drug metabolism in the main phase Ⅰ metabolizing enzyme cytochrome CYP450 enzymes, metabolic transformation of the early information to predict compound occurs in vivo, provide the basis for a potential drug safety and effectiveness. Identification of an important part of the analysis of metabolites in vivo metabolism studies, mainly through liquid chromatography-mass spectrometry techniques indeed signs of drug metabolites in vivo, revealing the body’s metabolism and excretion pathways in the transformation process. This thesis is divided into two parts, in vitro experiments to explore the metabolic properties of Salvia injection and in vivo experiments to explore and metabolic processes magnolol.Salvia injection of Salvia extract made from water-soluble formulation containing Danshensu, protocatechuic acid, a variety of ingredients protocatechualdehyde, Sal A and B, of which the Salvia higher pigment content. Due to its outstanding efficacy in heart and cerebrovascular disease, clinically increasingly widely used in conjunction with a variety of drugs, but Salvia injection on CYP450 enzyme relatively few research reports.Thus, the first part of this study, the use of probe drugs on Salvia Injection rat liver microsomes five kinds of CYP450 isoforms (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4) to explore the impact of using ultra performance liquid chromatography-tandem triple quadrupole mass spectrometry (UPLC-MS/MS) detection method to establish liver microsomal incubation system probe substrate metabolites, and its school system verification methods. The results show the specificity analysis established a strong, wide linear range, sensitivity, accuracy, repeatability, meet the general criteria for biological sample analysis methods can be used for quantitative analysis of target compounds. The results showed that Salvia injection on rat CYP1A2, CYP3A4 IC50 values> 100μg/mL, IC50 values for CYP2C9, CYP2C19 and CYP2D6 were 3.32μg/mL,16.44 μg/mL,0.30μg/mL. Salvia Injection on CYP1A2, CYP3A4 no inhibitory effect on CYP2C9 with moderate inhibition of CYP2C19 has a weak inhibitory effect on CYP2D6 showed a strong inhibitory effect. After the discovery of Salvia injection on rat liver microsomal enzyme CYP2C9, CYP2C19, CYP2D6 showing different degrees of inhibition, as compared species differences, and further study its effect on human liver microsomal enzymes, using probe drugs investigation human liver microsomal enzyme CYP2C9, CYP2C19, CYP2D6 impact injection Salvia, Salvia injection showed human CYP2C9, CYP2C19 and CYP2D6IC50 values> 50 μg/mL, showed no inhibition. The second part of this thesis research and honokiol in vivo metabolic properties. Honokiol are isolated from Magnolia officinalis medicine natural small molecule active substances, with a wide range of anti-tumor activity. Small intestine as an important organ of extrahepatic metabolism, its effect on the metabolism of drugs is equally important. In this paper, stable isotope labeling and ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) combined with technical, forensic analysis of metabolites in rat intestine and magnolol. First and 6 C atoms on the benzene ring Magnolol a stable isotope labeled with 13C, then mark with unlabeled honokiol preparation of 1: 1 mixture, by intravenous injection pulse rats (dosing dose:40 mg/kg), feces samples were collected and analyzed for each time period. Finally, in the small intestine were discovered 20 metabolites, which 20 metabolites are all two-phase metabolites, which are 12 amino acid conjugates, four sulfate conjugates,4 glucuronide conjugates,8 metabolites for the first time. For the system to understand and provide the basis honokiol metabolic pathways in vivo conversion. We first comprehensive analysis of the rat feces, plasma, intestinal metabolites, compare the difference of three matrix types and quantities of metabolites and magnolol and honokiol for further research in the metabolic processes in the body, systems characterize metabolic pathways provides a basis.