Discovery And Evaluation Of New EML4-ALK Inhibitors Against Non-small Cell Lung Cancer

Lung cancer,which is usually categorized into small cell lung cancer(SCLC,15%)and non-small cell lung cancer(NSCLC,85%),is the leading cause of cancer-related death all around the world.The expression of chimeric protein EML4-ALK is considered as one of the most important cause of NSCLC,thus makes EML4-ALK as a promising drug target in drug development.Since FDA approved Crizotinib as the first ALK inhibitor to use in clinic in 2011,Ceritinib,Alectinib,Brigatinib and Lorlatinib received approval by the FDA for the treatment of Crizotinib-refractory,ALK rearranged NSCLC.The point mutations of ALK kinase domain are considered to be one of the ’on-target’ mechanisms of aquired resistance after the use of ALK TKI.In this thesis,we initiated our work to discovery a new ALK inhibitor that is capble for inhiting all known resistant ALK point mutations.Firstly,ZZJ-03-087-01 was identified as a potent and highly selective EML4-ALK inhibitor.ZZJ-03-087-01 inhibited cell growth with IC50 values of 2.64 nM,12.03 nM and 49.04 nM against EML4-ALK-Ba/F3,EML4-ALK-L1196M-Ba/F3 and EML4-ALK-G1202R-Ba/F3,respectively,which is more potent compared with Crizotinib.And it showed great selectivity with S(1)=0.01 via KINOMEscan profiling.Molecular mechanism study revealed that ZZJ-03-087-01 specifically inhibited ALK signaling pathway which resulted cell apoptosis.Additionally,ZZJ-03-087-01 significantlly suppressed tumor growth in mouse xenograft models of H3122 and EML4-ALK-G1202R-Ba/F3 and genetically engineered mouse models of EML4-ALK and EML4-ALK-L1196M through tail vein injectin,but it’s not effective by oral dosage.Secondly,to improve oral bioavailability,ZBD-05-123-01 was identified through structural optimization.Systematical evaluation revealed that ZBD-05-123-01 exhibited superior antiproliferative activities with IC50S less than 1 nM against EML4-ALK-Ba/F3 and EML4-ALK-L1196M-Ba/F3,and 7.59 nM against EML4-ALK-G1202R-Ba/F3.ZBD-05-123-01 also had the selectivity score of S(1)=0.01 via KINOMEscan.More importantly,ZBD-05-123-01 was effective in all in vivo models tested via oral dosage.In summary,we have identified ZZJ-03-087-01 and ZBD-05-123-01 as novel,selective ALK inhibitors with great potency and high selectivity.Both of them were active against wt-EML4-ALK and drug induced EML4-ALK point mutations,which would serve as promising leads for new ALK targeted therapeutics to overcome the drug resistance in clinic.