Optimization Of Synthetic Route Of Anti-Gout Febuxostat And Synthesis Of Its Derivatives And Study Of Cysteine Bioluminescent Probes

In this thesis,the pathogenesis of gout and the mechanism of existing gout therapeutics are introduced briefly,and the recent advances of xanthine oxidase inhibitors(XOIs)in the treatment of hyperuricemia and gout are described in detail herein.Based on the crystal structure of xanthine oxidase,and the interaction modes of febuxostat as potent XOI and their structure-activity relationships,a series of phenylpyrimidine carboxylic derivatives were well designed through the modification of five-membered thiazole ring of febuxostat into a six-membered pyrimidine ring,the position change of carboxyl group on the pyrimidine ring,as well as the introduction of various substituted alkoxy groups at para-position of the benzene ring.With these efforts,we proposed to find new XOIs with better activity and weaker side effects.In the first part,a novel synthetic route for febuxostat with advantages such as the facile raw material,mild reaction conditions,convenient operations and high yield,was developed.Morevoer,starting from 2-hydroxybenzonitrile,eighteen substituted phenylpyrimidine carboxylic compounds were synthesized via alkylation,bromination,biphenylation,Suzuki coupling and hydrolysis,and confirmed by 1H NMR,13C NMR and HRMS.In vitro inhibitory activitites of target compounds were evaluated on bovine xanthine oxidase,and 7 compounds showed high xanthine oxidase inhibitory activities with IC50 ranging from 1.13 to 6.84 ?M.Compounds 16 and 18 exhibited the strongest inhibitory activities with 1.17 ?M and 1.13 ?M IC50,respectively.Nevertheless,they were not higher than the positive control Febuxostat(IC50?0.29 ?M).In the second part,two cysteine bioluminescent probes were well designed and synthesized.They were evaluated in vitro,cells,and transgenic mice levels.We found that the optimal incubation time of probe S-A and S-B is 10 and 100 min,respectively.In the case of bioluminescence intensity and cysteine concentration,there is a good linear correlation while the cysteine concentration is from 0 to 100?M.Moreover,the selectivity of probe S-B is higher than probe S-A.As for the bioluminescence intensity in interaction with cysteine,probe S-A and S-B is 20-and 34-fold higher than the blank group,6.5-and 5.5-fold than the homocysteine group,and 3.5-and 17-fold than the glutathione group,respectively.At the cellular level,we found that the bioluminescence intensity of probe S-B has a positive correlation with the concentration of cysteine.Forthermore,an extensive biological study of probe S-B was conducted at the transgenic mice level.The experimental rsults disclosed that probe S-B presented a good positive correlation with both endogenous and exogenous cysteine in mice.Therefore,this probe can be used to estimate the level of cysteine in living body.