Design, Synthesis And Antitumor Activity Of EGFR Kinase Inhibitor Based On T790M Target

Epidermal growth factor receptor(EGFR)kinases are key mediators of cell signaling involved in cell growth,proliferation,survival,and migration.Overexpression or mutation of EGFR kinases is a common feature of various human solid malignancies,particularly non-small cell lung cancer(NSCLC).Therefore,EGFR kinase has become a popular target in the research of anticancer drugs.In August 2002,Gefitinib was first marketed in Japan as the EGFR tyrosine kinase inhibitor for first-line treatment of non-small cell lung cancer.However,the first-generation EGFR inhibitors are limited by acquired drug resistance in clinical applications,while the second-generation EGFR inhibitors developed to solve drug resistance have the problem of poor selectivity for wild-type(WT)EGFR.Based upon the modeling binding mode of marketed AZD9291 with T790M,a series of 5,6-dihydro-4H-pyrrolo[3,2,1-ij]quinoline derivatives were designed and synthesized with the purpose to overcome the drug resistance resulted from T790M/L858R double mutations.And these EGFR inhibitors showed good activity and high selectivity for resistant tumors.A total of 20 target compounds not reported in the literature were synthesized.The structure of the target compounds were confirmed by high resolution mass spectrometry(HRMS),nuclear magnetic resonance spectroscopy(1H-NMR)and nuclear magnetic resonance spectroscopy(13C-NMR).The EGFR T790M/L858R kinase inhibitory activity test was performed on the target compound with AZD-9291 as a positive control drug.Among them,9 compounds with an IC50 value less than 20 nM were screened out and further antiproliferative effects were studied.The activities of A431,HCC827,and H1975 NSCLC cells carrying wild-type EGFR,EGFR delE746-A750,and EGFR L858R/T790M were measured.The results showed that the activity and selectivity of compound W3(A431:869.8 nM;HCC827:109.2 nM;H1975:102.6 nM)and positive drug AZD-9291(A431:615.6 nM;HCC827:61.6 nM;H1975:67.0 nM)were comparable.Further study can be conducted further.Microsomal stability tests in mice,rats,dogs,monkeys,and humans showed that compound W3 has a slightly shorter half-life than AZD-9291 and may have lower hepatotoxicity.In vivo anti-tumor activity studies showed that compound W3,when administered orally at 20 mg/kg/day for 18 consecutive days,significantly inhibited tumor growth in L858R/T790M-driven human NSCLC xenograft mice.In addition,the binding affinity of compound W3 to the hERG ion channel was lower than that of AZD-9291,indicating that the candidate compound may have less cardiotoxicity than AZD-9291.In vitro and in vivo data indicate that compound W3 can be developed as a clinical candidate.