海洋真菌次生代谢产物鹿角缩酮B对大鼠CYP3A2的作用分析-药理学专业论文.docxVIP

暨南大 暨南大学硕士学位论文 PAGE PAGE IV Objective: Abstract Through computer-aided molecular docking method, in vitro liver microsomal incubation method and pharmacokinetic studies, aim to evaluating the ability of Xyloketal B to inhibit the P450 enzyme CYP3A ,predicting drug-drug interaction and providing experimental evidence for clinical research. Methods: Using computer-aided molecular docking and the scoring function to judge space binding capacity between Xyloketal B and major CYP450 isoforms. Midazolam as an probe substrates, liver microsomes in vitro incubation method was used to investigate the potency and mechanism of Xyloketal B on rat hepatic CYP3A and and its mechanism. Using midazolam as a probe drug,to study Xyloketal B on the pharmacokinetics of midazolam and its major metabolite 1-hydroxy midazolam in rats ,meanwhile analyze the impact Xyloketal B on on rat hepatic CYP3A with western blot method and P450-GloTM kit method. Results: Evaluation Xyloketal B and major CYP450 isoforms binding capacity by the scoring function.Total-score results were as follows: CYP3A4-3NXU:6.0461; CYP11B2-4DVQ: 5.0078;CYP2D6-3TBG:4.9342;CYP2C9-1R90:3.4475;CYP1A2-2HI4:-1.2051;CYP2E1-3 T3Z:-8.3081;CYP2A6-2FDV:-16.2292; Binding capacity:CYP 3A4 CYP11B2 CYP2D6 CYP2C9 CYP1A2 CYP2E1 CYP2A6. Prompting Xyloketal B with a maximum binding capacity of CYP3A4. The results of enzyme kinetics show that Xyloketal B as rat liver microsomes weak CYP3A inhibition, IC50 was 27.91 μmol/L, Ki , Ki, KI and Kinact respectively 41.85 ,31.67, 0.063 μmol/L and 0.0037min-1. Xyloketal B is a mixed type of non-competitive-anti-competitive inhibitor and with NADPH, time and concentration dependent manner.This result suggested that Xyloketal B may nihibit the activity of CYP3A through mechanism-based mecheanism. The results showed that compared with normal saline or soybean oil, ketoconazole increased both AUC0-t and Cmax , as well as reduced CL/F and AUC0-inf ratio of 1-OH MDZ and MDZ, which were st