Design,Synthesis And Preliminary Antitumor Activity Of Novel Diphenylaminopyrimidine Compounds

Objective:A series of novel diphenylaminopyrimidine small molecule compounds were designed and synthesized,and their anti-tumor pharmacological activities were preliminarily studied in order to obtain small molecule drugs with better anti-cancer activity and lower toxicity,especially for pancreatic cancer.Methods:Based on the previous structural modification of pyrimidine heterocyclic anti-cancer drug Rociletinib and the basic pyrimidine ring skeleton was retained,the acrylamide group bound to the Michael addition receptor was replaced with other conjugated groups.The C-5 atom of the pyrimidine ring was replaced with a chlorine atom,and other different side chains were introduced to the C-2.Finally,19diphenylaminopyrimidine compounds in three series were synthesized,and the structures of the targeted compounds were confirmed by ¹H NMR,¹³C NMR and HRMS.Subsequently,we performed their biological evaluation.Gemcitabine was used as a positive reference drug.CCK-8 method was used to detect the half inhibition concentration IC₅₀ of the targeted compounds on human normal liver cells（L-02）,human non-small cell lung adenocarcinoma cells（H1975）,human metastatic pancreatic adenocarcinoma cells（As PC-1）,human colon cancer cells（HCT-116）,human non-small cell lung cancer cells（H322）and other cancer cell lines within 72 h.According to the results of biological evaluation experiments,the targeted compounds with the best inhibitory activity on As PC-1 cells and less cytotoxicity on L-02 cells were selected,and then the apoptosis rate of pancreatic cancer cells As PC-1 was investigated by AO/EB double staining and DAPI staining.Flow cytometry was used to investigate their ability to induce apoptosis of As PC-1 cells and the effect on cell cycle.The effect of the optimal targeted compound on the proliferation and migration of pancreatic cancer cell As PC-1 was evaluated by cell scratch assay.Results:¹H NMR、¹³C NMR and HRMS datas showed that the targeted molecular structures were correct.The results of CCK-8 assay showed that most of the compounds had significant proliferation inhibitory effects on a variety of cancer cells,especially on the pancreatic cancer cells As PC-1.Therein,compound ZO-5 had the best activity on As PC-1 cells and and the lowest toxicity on normal liver cells L-02,which was much better than gemcitabine 15 times.The results of AO/EB double staining and DAPI staining showed that the number of pancreatic cancer cells treated with ZO-5 was significantly reduced,and the nucleus was condensed,showing significant apoptosis.Flow cytometry results showed that compound ZO-5 could significantly induce apoptosis of As PC-1 cells and the cell proliferation was arrested in G2/M phase.The results of cell scratches assay showed that compound ZO-5 could inhibit the migration ability of As PC-1 cells,which was also better than gemcitabine.Conclusion:The new diphenylamine pyrimidine skeleton compounds synthesized in this study could significantly inhibit the proliferation of pancreatic cancer cells.Therein,the optimal compound ZO-5 had a better inhibitory effect on pancreatic cancer cells than the positive reference drug gemcitabine,and had higher selectivity for pancreatic cancer cells and lower toxicity to normal liver cells.It had the significance and value of in-depth development.