Synthesis And Anti-Pancreatic Cancer Activity Of Novel Pyrimidine FAK Inhibitors

Objective:In this study,new pyrimidine compounds were designed and synthesized according to the lead compound TAE226,and there anti-pancreatic cancer activity was preliminarily evaluated,so as to find new anti pancreatic cancer FAK inhibitors with low toxicity,good selectivity and good activity.Methods:Based on the structural characteristics of FAK,11 new FAK inhibitors were designed and synthesized by analyzing the structure-activity relationship between TAE226 and CT707,introducing isopropyl sulfonyl to the diarylamine pyrimidine skeleton and changing the C-2 amino side chain structure.Series compounds were identified by NMR and HRMS.First,with TAE226 as positive reference drug,the inhibitory activity of FAK was determined by ADP-Glo^TMmethod;The inhibitory effects of the new compound on human pancreatic cancer cell lines AsPC-1,PaCa-2,human lung cancer cell line H1975 and bortezomib resistant multiple myeloma cell line KM3/BTZ were tested by CCK-8 method.In addition,the hepatotoxicity of the compound was evaluated by human normal liver cell line L-02.The effect of the target compound on the migration ability of AsPC-1 cells was evaluated by scratch test.The cell nucleus and apoptosis were observed by DAPI staining and AO/EB fluorescence staining.In order to further study the antitumor mechanism of the target compound,flow cytometry was used to study the apoptosis mechanism.Finally,animal models were established to investigate the antitumor activity of the optimal compound in vivo.Results:In this study,11 new pyrimidines were designed and synthesized,and their structures were characterized correctly by high-resolution mass spectrometry and nuclear magnetic resonance.Preliminary pharmacological activity test results indicated that most of the compounds had good pharmacological activity.The results of FAK activity showed that most of the compounds had strong inhibitory activity against FAK.FAK enzymatic activity at concentrations ranging from 0.1165 to 7.553 n M,among which compound 9f has the strongest inhibitory activity against FAK(IC₅₀=0.1165μΜ).The results of anti-proliferation showed that most of the compounds had strong anti-proliferation effects on various malignant tumor cells（AsPC-1,PaCa-2,H1975 and KM3/BTZ）,even better than the positive reference drug TAE226.Compound 9f showed strong antiproliferative activity on AsPC-1 and PaCa-2 cells,with IC₅₀of 0.1596μM and 0.7679μΜ,but had relatively low toxicity to L-02(IC₅₀=1.398μΜ).The results of scratch experiment showed that compound 9f effectively inhibited the migration ability of AsPC-1 cells.DAPI staining and AO/EB fluorescence staining showed that compound 9f effectively inhibited the proliferation of AsPC-1 cells.Flow cytometry showed that compound 9f inhibited the cell cycle of AsPC-1 cells in G2/M phase.In vivo experiments with xenogeneic mouse model further showed that compound 9f also had good antitumor effect in vivo.Conclusions:This study designed and synthesized 11 new pyrimidine compounds.Most of compounds had good activity against FAK activity and tumor cell.At the same time,the activity of the optimal compound 9f was significantly better than that of the lead compound TAE226 in vitro,and in vivo experiments showed that it had a significant inhibitory effect on pancreatic cancer.In conclusion,compound 9f may be a potentially better drug for pancreatic cancer,and it is worth further structural optimization and pharmacological activity research.