Establishment Of Mouse Hyperuricemia Model And Design,Synthesis And Activity Evaluation Of Lesinurad Derivatives

Gout is a common metabolic disease caused by disorder of purine metabolism and/or under-excretion of uric acid.Its pathological conditionsis hyperuricemia,which is associated with several clinical features such as acute gouty arthritis,gout nodules,chronic arthritis,joint deformities,chronic nephritis and uric acid nephrolithiasis.The prevalence of gout and hyperuricemia is increasing worldwide over the past few decades,bringing tremendous burden to patients and society.Currently,the clinical drugs for treating gout mainly include traditional anti-inflammatory drugs(gout inflammatory interference),xanthine oxidase inhibitors(inhibition of uric acid production)and uricosuric drugs(promoting uric acid excretion).There are still unmet clinical needs in China and othercountries.Anti-gout research has gradually become a hot field of drug development,and a variety of new chemical entities,novel therapeutics have emerged in recent years.Among them,uric acid transporter 1(URAT1)is an attractive target of anti-gout drug research.Lesinurad(RDEA594)was the first URAT1 inhibitor approved in 2015 by the FDA,and the combinations of xanthine oxidase inhibitors(XOI)with Lesinurad can safely decrease urate levels in gout patients.Thus,Lesinurad provides new treatment options for gout patients who are ineffective with XOI,and opens up new areas of anti-gout drug development.The establishment of drug screening method is a basic work in the process of developing pharmaceutical products.Establishing animal models for screening bioactive compounds has been widely used by researchers.Animal models of hyperuricemia is the key to studying the pathogenesis of this kind of disease and relative therapydrugs.There are many methods to establish hyperuricemia models,but most of them are limited in the evaluation of novel anti-gout compounds because of the low stability and poor repeatability.So in this study we established a relatively stable mouse model of hyperuricemia by analyzing the deficiencies of the existinganimal models of hyperuricemia and combining the previous work of our research group.Additionally,the safety of the model for liver and kidney function was also evaluated.The specific method was to give hypoxanthine(precursors of uric acid)and potassium oxalate(uricaseinhibitor)to mice in a single large dose for a short time to reach high levels of uric acid.The single variable method was used to study the effects on models uric acid levels from the three influence factors,including methods of administration,dosage of modeling drugs and blood collection time.In the end,levels of glutamic oxalate transaminase(AST),alanine transaminase(ALT),serum creatinine(SCr)and blood urea nitrogen(BUN)were used to evaluate the function of liver and kidney of the model animals in order to investigate the safety of the model,and finally a safe and stable mouse model of hyperuricemia was obtained.Lesinurad offers new medication options for the treatment of gout,but it has some shortcomings,such as high liver and kidney toxicity,and narrow therapeutic window.So it is still necessary to carry out further structural modification.In this study,two series of novel compounds were designed and synthesized by using different drug design strategies,such as scaffold hopping,bioisosterism and so on.In the first series,sixteen N-acyl sulfonamide compounds were synthesized,the carboxyl group in the structure of Lesinurad was replaced by N-acyl sulfonamide group.Meanwhile,alkyl,phenyl,substituted phenyl and thiophene groups were introduced into the structure to increase the structural diversity and explore the structure-activity relationship,so as to provide guidance for further study.In the second series,the side chains of mercaptoacetic acid in the structure of lead compounds were retained,whilethetriazole group was replaced with uric acid-like structures such as benzopyrimidinone and pyridinopyrimidine,by employing scaffold hopping principle,whichresulted in twelve newly designed and synthesized Lesinurad derivatives.The in-house established hyperuricemia mouse model was used to evaluate the anti-gout activity.Among the 28 compounds,more than 10 compounds were found to be more active than the positive control,which validated the rationality of our design idea.These Lesinurad analogues could be promising lead compounds for further optimization.In conclusion,a stable,reliable,safe and economically viable model of hyperuricemia in mice has been established,which provided an effective and rapid screening method for anti-gout drug research.Besides,in this thesis,we designed,synthesized and tested the anti-gout activity of Lesinuard analogues in vivo.The results suggested that more than 10 novel compounds have higher activity in reducinguric acidthan Lesinurad,which showed a potential value in further development.In addition,the structure-activity relationships were summarized to provide reference for the further structural optimization.