Experimental Study On The Characteristics Of Compound CC15009 For Anti-hyperuricemia Based On XOR And Its Subtypes XO、XDH

Xanthine oxidoreductase(XOR),the key enzyme catalyzing the production of uric acid(UA)from purine,consists of two subtypes in vivo,xanthine dehydrogenase(XDH)and xanthine oxidase(XO).In the catalytic reaction of XDH or XO,NADH or ROS is generated as byproduct,respectively.This study aims to establish the methods for the activity determination of XOR and its subtypes,to investigate the characteristics of the inhibition of compound CC15009 on XOR and its subtypes,respectively,then to explore the therapeutic effect of compounds on hyperuricemia based on the target XOR,in vivo.Firstly,based on the different electron acceptor of XDH or XO,the methods for measuring the activities of XOR,XDH,or XO from bovine milk(pure enzyme)and mouse liver were established,respectively.The optimal concentrations of the substrate and electron acceptor,pH value,reaction time,were determined,and range of the enzyme activity which could be well detected was investigated,respectively.Subsequently,the interaction characteristics of compound CC 15009 with XOR and its subtypes were studied utilizing molecular docking,surface-plasmon resonance test,and enzyme kinetics..Finally,the anti-hyperuricemic effects of the compound based on its inhibition on XOR were observed in three different hyperuricemic mouse models.In conclusion,we successfully established new methods for determining the activity of XOR and its subtypes,XDH and XO.We explored the inhibitory characteristics of compound CC15009 on XOR and its subtypes,and investigated its in vivo anti-hyperuricemic effects by XOR inhibition.