Design,Synthesis And Antitumor Activity Screening Of ALK/EGFR Dual-target Inhibitors

Lung cancer accounts for about a quarter of global cancer deaths,and the 5-year survival rate is less than 20%.The most common cancer subtype in lung cancer is non-small cell lung cancer（NSCLC）,which accounts for approximately 80-85%of total cases.Anaplastic lymphoma kinase（ALK）and epidermal growth factor receptor（EGFR）are well-recognized effective targets for the treatment of non-small cell lung cancer.Targeted inhibition of ALK and EGFR can effectively improve the life quality of NSCLC patients and prolong their survival time,but the acquired drug resistance limits the therapeutic effects of traditional ALK and EGFR single-target inhibitors more or less.Dual-target inhibitors block two synergistic pathways simultaneously which inhibits tumors more potent and delays the emergence of acquired resistance.Therefore,it is a more effective way to develop ALK/EGFR dual-target inhibitors.Currently,Briagtinib is the only marketed ALK/EGFR dual-target inhibitor,which showed a good therapeutic effect in patients with triple EGFR mutations,but a diminished therapeutic effect in patients with ALK G1202R mutations.To overcome the resistance of ALK G1202R and enhance the inhibitory effect against triple mutant EGFR kinase,our group intends to develop a novel ALK/EGFR inhibitor.We have already obtained an ALK inhibitor ZH-1 with a similar chemical structure compared to Brigatinib.In the thesis,the binding mode of Brigatinib and ALK kinase（PDB code:4CLI,6LUB）is studied to conduct the discovery and optimization of ZH-1.We retained the 2,4-diamino-5-chloropyrimidine core for the good inhibitory potency of ZH-1 on ALK and modified it with a series of branched chains using computer-aided drug design.Then,compounds were screened according to the principles of easy availability of raw materials,mild reaction conditions,and high molecular docking scoring,and 167 compounds were obtained.Among them,35 compounds with novel structure have been synthesized by chemical means and identified by HRMS and 1H-NMR.In this thesis,it was found most of our compounds can inhibit ALK and EGFR kinases by more than 90%at a concentration of 10μM by kinase assay in vitro,which indicates that our design strategy of ALK/EGFR inhibitor is reasonable.Then the 35compounds are taken into cell assay by CCK8 method to screen the anti-tumor proliferation activity on PC-9-LRTM,Karpas-299,and H1975 cell lines.The results of the cell assay showed that most of the 35 compounds had a good inhibitory effect on cancer cells with ALK or EGFR mutation.Among them,the antitumor activities of compounds AE-9,AE-22,and AE-167 were comparable to the marketed drug Briagtinib.The IC₅₀ of AE-25 on PC-9-LRTM was 249.8 n M,which was higher than that of Briagtinib(IC₅₀=834.5 n M)by 3 times.And,the IC₅₀ of AE-42 against karpas-299 was80.2 n M,which was 2 times better than that of Briagtinib(IC₅₀=169.4 n M).Both AE-25and AE-42 are of good antitumor effects in vitro,which deserve further study.