We have done large throughput inhibitor testing against multiple cancer associated tyrosine kinase objectives and obtained a few visits with previously unreported chemical scaffolds. On the list of types we found a compound that strongly order Dizocilpine inhibits ALK, consequently, we intensively revised and enhanced its kinase selectivity and pharmacokinetics report. Eventually, we identified a benzo carbazole derivative, CH5424802, as a potent, selective, and orally available ALK chemical. In cellfree assays the IC50 of CH5424802 for enzyme action of ALK was 1. 9 nM, the dissociation constant value for ALK in an ATP competitive way was 2. 4 nM using a competition binding assay. The inhibitory activity for two hot spot activating mutations in neuroblastoma, F1174L and R1275Q, was much like that for wildtype ALK. To examine the kinase selectivity of CH5424802, its inhibitory action on different kinases was measured, revealing weak or no inhibition against 24 protein kinases other than ALK. More over, applying Ambits kinase Eumycetoma testing system, CH5424802 was profiled against 402 kinases like the mutated kinases. Only three kinases, ALK, GAK, and LTK, showed over 50 inhibition at 10 nM, which corresponds to about 5 fold higher concentration of IC50 values for ALK. LTK is well known to show best sequence similarity to ALK. In cellular phosphorylation assays, CH5424802 might reduce autophosphorylation of ALK in NCI H2228 NSCLC cells revealing EML4 ALK, and it also triggered substantial elimination of phosphorylation of STAT3 and AKT, but not of ERK1/2. However, inhibition of these phosphorylations was not noticed in ALK mix negative A549 cell line. In NSCLC, EML4 ALK fusion has been shown to be mutually exclusive with EGFR or KRAS strains. It’s already been reported that EGFR tyrosine kinase inhibitors have high clinical efficacy as therapeutic agents for NSCLCs with natural compound library EGFR versions. Using NSCLC cell lines with distinct genotypes, we examined the effects of CH5424802. CH5424802 was preferentially effective against NCI H2228 cells showing EML4 ALK, however not ALK mix bad NSCLC mobile lines, including HCC827 cells, A549 cells, or NCIH522 cells in monolayer culture. Similar results were obtained under 3d spheroid culture conditions. CH5424802 can induce an marker?caspase 3/7 like initial? in NCI H2228 spheroid cells, indicating the contribution of apoptosis induction in the antitumor activity of CH5424802. The caspase 3/7 like service was also observed in remedy of other ALK inhibitors, PF 02341066 and NVPTAE684, under spheroid culture conditions. Also, we proved that EGFR mutant HCC827 cells showed higher sensitivity to the EGFR kinase inhibitor gefitinib, however not to CH5424802.