Design、Synthesis And Biological Evaluation Of EGFR Inhibitors And Researh On The Synthesis Of Polysubstitued Pyrazoles

Malignant tumor is one of the maior diseases that seriously threat human’ s health.At present,chemotherapy for malignant tumor is gradually changing from traditional cytotoxic drugs to targeted therapy.Epidermal growth factor receptor tyrosine kinase(EGFR)is an important target of tumor therapy.Anti-tumor drugs based on EGFR have undergone three generation inhibitors.Desensitization and drug resistance occurred in some patients after they receive the treatment of the first generation inhibitors.To some extent,the second generation inhibitors and the third generation inhibitors alleviated the problem.HDAC inhibitors could change the acetylation level of histones,and then regulate gene transcriptional processes,resulting in extensive antitumor activity.Multi-target ed inhibitors based on HDAC inhibitors are better than single target drugs,and can reduce the drug resistance.Through combination principles,key motif of HDAC inhibitors were fused to the second generation irreversible EGFR inhibitors to build EGFR/HDAC double-acting inhibitors.We tested the antiproliferation activity of the 14 synthesized compounds in vitro,the results show that some compounds showed good antiproliferation activity on tumor cell lines A431、H1975、HeLa and HepG2.Among them,the activity of H-5 is relatively better.Flow cytometry analysis showed that compound H-5 has induced the apoptosis of H1975 cell in a dose-dependent manner.Western Blot experiment showed that the ability of H-5 to inhibit phosphorylation of EGFR was less weaker than that of Afatinib.Subsequent activity study of this kind of compounds are still in process.Our previous study showed that aziridine is a mild covalent adduct fragment,could replace acrylamide to conduct Michael addition reaction with EGFR protein.So,based on the third generation EGFR inhibitors,we introduced aziridine ring into pyrimidine core and constructed a series of aziridine containning pyrimidine derivatives.We evaluated the antiproliferation activity of the synthesized 30 compounds in vitro.The results showed that the compounds had good selectivity for L858R/T790M double mutant cell line H1975,which was consistent.with the positive WZ4002.We further tested the kinase activity of seven compounds,the results showed that although this kind of structure kinase inhibition ability were lower than the positive WZ4002 and CO-1686,they still has good selectivity for the L858R/T790M double mutant EGFR kinase.Flow cytometry analysis showed that compound HW-19 had induced the apoptosis of H1975 cell in a dose-dependent manner.Additionally,HW-19 is relatively stable in the S9 Metabolic system.A method for the synthesis of polysubstituted pyrazoles has also been developed.polysubstituted 4-aminopyrazoles and 4-hydroxypyrazoles were synthesized from vinyl azides and hydrazines.The reactions appear to be useful and covenient due to readily available starting materials,experimental simplicity and mild reaction conditions.