Design,Synthesis And Biological Evaluation Of Urea(Thiourea)-Substituted 2-anilinopyrimidine Derivatives As Nur 77 Regulators

Nur77 belongs to the NR4A subgroup of the nuclear receptor superfamily,and its diverse functions such as promoting apoptosis,promoting cell proliferation,affecting angiogenesis and cell migration make it an important target for anticancer drug development.In addition,2-anilinopyrimidine is an important structural unit.Natural compounds or synthetic compounds containing this structural unit have a rich biological activity,and it has a wide range of applications in the field of medicine,which makes it become one of the research hot spots of organic chemists and drug chemists.Based on the previous work of the research group,we designed a series of urea?thiourea?-substituted 2-anilinopyrimidine compounds.Through a 5-step synthesis route,a total of 79 2-anilinopyrimidine compounds were obtained.The structures of all the compounds were confirmed by ¹H NMR,¹³C NMR and HRMS spectra.The preliminary biological activities of the synthesized 2-anilinopyrimidine compounds were evaluated by means of MTT,Western Blot,cell transfection and immunofluorescence.The experimental results are as follows:?1?Most of the compounds shows good cytotoxic activity,and the cytotoxic activity of A-12 was the best at 0.61 ?M.The preliminary structure-activity relationship showed that the biological activity of thiourea compounds was slightly better than that of urea compounds.When the para-pyridine is introduced into the four-position of pyrimidine,its cytotoxic activity is better then others.On the other hand,the para-position substitution on the phenyl ring of the 2-anilinopyrimidine makes it a better structure than the meta-position substitution,which contain better biological activities.In the choice of R2 substituents,benzyl and halogen-substituted phenyl rings shows better effects,and aromatic hydrocarbons are superior to aliphatic hydrocarbons.;?2?The results of Biacore test showed that compound A-12,A-14,A-16,A-26,A-34 can bind with Nur 77 better than others;?3?In the activity screening experiment of inducing apoptosis,some of the compounds?A-1,A-5,A-6,A-12 et.?showed excellent performance,which had a good activity of inducing cell cycle arrest and apoptosis,and could down-regulate the expression of Nur 77;?4?Studies on compound A-12 have shown that A-12 has a good ability of inducing apoptosis,and this ability has a certain concentration-and time-dependent,and a certain degree of selectivity for cell lines.It works better in liver cancer cell lines.A-12 has a strong ability to down-regulate the expression of Nur 77 protein.Further experimental results show that its ability to induce apoptosis is related to Nur 77.