Preliminary Study On The Synthesis And Anti-tumor Activity Of 2-substituted-4-arylaminopyrimidines

In the world,especially in China,cancer seriously threatens people's health and quality of life.With the development of biology and related technologies,the treatment of cancer has reached the molecular levels,and targeted therapy has become the direction of the development of cancer treatment,which brings hope to the cancer patient.Pyrimidine and its derivatives have been reported with high values in targeted therapy of cancer.4-Arylaminopyrimidine derivatives,such as Osimertinib and Imatinib etc,are usded as tyrosine kinase inhibitors(TKIs)and showed the better effect in the early stage of treatments in clinic.Unfortunately,after the treament with such TKIs for 9-14 months,the most of patients obtained drug-resistance.Therefore,it is extremely urgent to design and synthesize new anti-tumor drugs with high acticity,low toxicity and low drug resistance.In this paper,in order to find new anti-tumor compounds with high activity and low toxicity,a series of 2-substituted-4-arylaminopyrimidines were designed by using computer-aided drug design software Sybyl,then chemically synthesized.The main contents of this study are as follows:1.Based on the review of the progress in the treatment of cancer,especially molecular targeted therapy,the progress in the study of biologically active pyrimidines and their derivatives,and the development of pyrimidine-based tyrosine kinase inhibitors,herein,Osimertinib and EGFR(PDB No:4LL0)were usded respectively as reference ligand and receptor,via molecular docking by virtual screening with computer-aid drug design software Sybyl,fifteen 2-substituted-4-arylaminopyrimidines were selected as target compounds,and then were chemical synthesised.2.For synthesis of target compounds,firstly,an important kind of intermediate 2-chloro-4-arylaminopyrimidine compounds were synthesized.The nucleophilic substitution of 2,4-dichloropyrimidine and five aromatic amines,including 3-chloro-4-fluoroaniline,3-ethynylaniline,3-chloro-4-(3-fluorobenzyloxy)-aniline,3-chloro-4-(pyridin-2-ylmethoxy)aniline and 3-chloro-4-(3-(trifluoromethyl)-phenoxy)aniline were carried out,respectively.The substitution reactions occurred selectively in the 4-position of 2,4-dichloropyrimidine by using Et3N as acid-binding agent and ethanol as solvent.Thus,five intermediate compounds,including 2-chloro-4-(3-chloro-4-fluorophenylamino)pyrimidine,2-chloro-4-(3-ethynylphenylamino)pyrimidine,2-chloro-4-(3-chloro-4-(3-fluorobenzyloxy)phenylamino)pyrimidine,2-chloro-4-(3-chloro-4-(pyridin-2-ylmethoxy)phenylamino)pyrimidine and 2-chloro-4-(3-chloro-4-(3-(trifluoromethyl)phenoxy)phenylamino)pyrimidine,were successfully prepared in 65-68%yield.3.Five "N" series of target compounds C1-C5 were synthesized by nucleophilic substitution reaction between 2-diethylaminoethylamine and five 2-chloro-4-arylaminopyrimidine intermediates respectively in 66?82%yield.Five "O"series of target compounds C6-C10 were also synthesized in 50?58%yield by nucleophilic substitution between 2-diethylaminoethanol and five 2-chloro-4-arylaminopyrimidine intermediates.Finally,five "S" series of target compounds C11-C15 were synthesized in 62?66%yield by using five 2-chloro-4-arylaminopyrimidine intermediates and the isothiouronium salt of N,N-diethylaminochloroethane hydrochloride as raw mateials.The structures of fifteen synthesized target compounds were confirmed by 1H NMR,13C NMR,HRMS and IR.4.The antiproliferative activities of the fifteen synthesized 2-substituted-4-arylaminopyrimidine compounds were evaluated by MTT assay in vitro.The clinical anti-tumor drugs Lapatinib,Gefitinib and Osimertinib were used as positive controls,and human colon cancer cells SW480,human squamous cell carcinoma A431,human non-small cell lung cancer cells A549 and NCI-H1975 were used as test cells.The results showed that C3,C5,C6,C13 and C15 showed good antiproliferative activities against four tested tumor cells,and their IC50 values were 0.36?6.24 ?M,which was better than clinical drug Gefitinib and Lapatinib.In particular,compounds C3 and C13 showed potent antiproliferative activities against four tested tumor cells.The IC50 values were in the range of 0.36?3.48 ?M and 0.40?1.88 ?M,respectively,which was equivalent to the activity of clinical drug Osimertinib(0.05?2.67 ?M).Compound C3 was further evaluated for its effect on the apoptosis and cycle of tumor cells.The results showed that compound C3 could induce the apoptosis and arrest cell cycle in G0/G1 phase when A549 tumor cells were treated by C3 for 48 hours.