Synthesis And Anti-cancer Activity Evaluation Of Novel Pyrimidine Derivatives As EGFR Inhibitors

Human epidermal growth factor receptor（EGFR）is an important drug target that plays a fundamental role in signal transduction pathways in oncology.Synthesis of novel EGFR tyrosine kinase inhibitors has become a hotspot of pharmaceutical research,and pyrimidine derivatives are widely distributed in the human body and other living body as an important class of life substances.Compounds containing a pyrimidine skeleton have received significant attention in medicine,agriculture and other fields as this scaffold has broad biological activities,such as antitumor,insecticidal activities.In this thesis,we report herein the discovery of a novel class of pyrimidine derivatives with improved potency toward EGFR.Means of Computer Aided Drug Design was utilized,and the activities of the designed compounds were evaluated.The results were concluded as follows:The intermediate 2-amino-4,6-dihydroxypymidine was first synthesized with diethyl malonate and guanidine nitrate as raw materials by reaction of nucleophilic addition-elimination reaction（cyclization）in sodium ethylate.Then,2-amin-4,6-dimethoxypyrimidine was synthesized by chlorination reaction and alkoxylation reaction.Finally,the 18 novel N-（4,6-dimethoxy-pyrimidin-2-yl）-2-（4-phenyl-piperazin-1-yl）acetamide derivatives（3a-3r）were obtained by the connection of 2-amin-4,6-dimethoxypyrimidine and various substituents piperazine.All of the compounds were characterized by ¹H NMR and MS spectra.Besides,two compounds were structurally determined by single crystal X-ray structural analysis.The biological activities of all the synthesized compounds were tested,and we discussed the structure-activity relationships were discussed.The results showed that all of the compounds had potential anticancer and EGFR inhibitory acitivities.Some compounds exhibited excellent activities with low toxicity,comparable to the positive control Erlotinib(IC₅₀=0.15μM for MCF-7,IC₅₀=0.11μM for Hela,IC₅₀=0.10μM for A549).Compound 3p showed the best anti-proliferative activity(IC₅₀=0.22μM for MCF-7,IC₅₀=0.08μM for Hela,IC₅₀=0.05μM for A549)and inhibitory activity on EGFR(IC₅₀=0.02μM).Compoutational docking studies also showed that compound 3p has interaction with EGFR key residues in the active site.Particularly,the binding energy between compound 3p and EGFR was-48.81 Kal/mol.Meanwhile,the hydrogen bonding,π–π and π–sigma interactions between EGFR activity center and compound 3p provide a strong theoretical support for pymidine derivatives as EGFR TK inhibitors.