Anti-hyperuricemia Pharmacodynamics Of Anemoside B4 And Pharmacokinetics Of Different Administration Routes

Objective:To investigate the pharmacodynamic effect of Anemoside B4(AB4)on hyperuricemia(HUA),and make a preliminary exploration of its uric acid-lowering mechanism.Meanwhile,to explain the pharmacokinetics of AB4 in rats through different administration routes,which provides the basis for the preliminary research of its later-stage finished drug research.Methods:1.The hyperuricemia model of acute ICR mice was constructed by intraperitoneal injection of 450 mg/kg potassium oxonate,and AB4 was pre-administered for one week to preliminarily investigate the preventive effect of AB4 on hyperuricemia2.The hyperuricemia model of KKAy mice was constructed by feeding with high uric acid special feed,and administered at the same time as the model was established,to further investigate the drug effect of AB4 on hyperuricemia.3.Through the substrate-enzyme incubation experiment in vitro,the inhibitory effect of AB4 on xanthine oxidase was determined by enzyme labeling instrument.4.Based on HPLC-MS/MS technique and purine metabolic pathway,a targeted metabonomic analysis method for the determination of uric acid and its precursors in mouse liver was established to detect the effect of AB4 on the content of uric acid and its precursors in the liver of KKAy mice.5.The gene expression levels of uric acid transporter OAT1,OAT3,BCRP,URAT1,GLUT9 and NPT1 in liver and kidney of two hyperuricemia model mice were detected by real-time fluorescence quantitative PCR.6.A high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)method established in the laboratory and verified by methodology was used to investigate the pharmacokinetic differences of rats after intragastric administration of AB4 solution,enteric-coated capsule,rectal administration of AB4 suppository and tail vein injection of AB4 solution.Results:1.In the acute hyperuricemia model of ICR mice,the serum uric acid level of the model group mice was significantly higher than that of the blank group mice,and the hyperuricemia mouse model was successfully constructed.Pre-administration of AB4 can effectively reduce the blood uric acid level of mice in the acute hyperuricemia model of ICR mice.2.In the hyperuricemia model of KKAy mice,the levels of serum uric acid,blood urea nitrogen and triglyceride in the model group were significantly higher than those in the blank group,and the hyperuricemia model was successfully established.Anemoside B4 can effectively reduce the levels of serum uric acid and urea nitrogen in KKAy mice with hyperuricemia.3.AB4 can inhibit the activity of xanthine oxidase to some extent in vitro.4.AB4 can reduce the content of uric acid precursor in the liver of KKAy mice,thus affect the production of uric acid in the liver and reduce uric acid.5.AB4 can up-regulate the levels of OAT3 m RNA,BCRP m RNA and NPT1 m RNA in the kidney of ICR mice with acute hyperuricemia,up-regulate the level of BCRPm RNA in the liver,and down-regulate the expression of URAT1 m RNA in the kidney,thus affecting uric acid metabolism and reducing uric acid.6.AB4 can up-regulate the levels of OAT1 m RNA,OAT3 m RNA,BCRPm RNA and NPT1 m RNA in kidney tissue of KKAy mice with hyperuricemia,and up-regulate the level of BCRP m RNA in liver tissue,thus affecting uric acid metabolism and achieving the purpose of reducing uric acid.7.Compared with the pharmacokinetic parameters of oral AB4 solution,enteric-coated capsule and rectal suppository,the exposure of rectal AB4 suppository in rat plasma was 49 and 28 times higher than that of oral AB4 solution and enteric-coated capsule at the same dose.Conclusion(s):1.AB4 can significantly reduce the level of serum uric acid in hyperuricemia model mice.On the one hand,AB4 inhibits the activity of xanthine oxidase and the production of uric acid precursors,thus reducing the production of uric acid.On the other hand,AB4 regulates the expression of uric acid transporter OAT1,OAT3,BCRP,NPTI and URAT1 and affects uric acid transport and excretion.2.The bioavailability of AB4 is very low after oral administration.AB4 rectal suppository and enteric-coated capsule can increase its exposure in vivo to some extent,but rectal suppository has the most significant effect,so it can be used as a potential dosage form to improve the bioavailability and exposure of AB4 in vivo.