LAUSR

Aiming to develop ALK/ROS1 dual inhibitors overcoming ceritinib-resistant G1202R mutant, a dedicated structure-guided modification campaign was conducted based on ALK co-crystal structures. Twenty eight diarylaminopyrimidine (DAAP) analogues possessing furan or tetrahydrofuran group were designed and synthesized, among which compound 16 bearing (dimethylamino)methyl)furan-2-yl)methyl)thio fragment was identified. Compound 16 exhibited significant cytotoxicity on ALK-positive Karpas299 and H2228 cells with IC50 values of 20 nM and 110 nM. Meanwhile, compound 16 turned out as the most potent entity superior to ceritinib with IC50 values of 2.8, 2.6, 3.8 and 2.3 nM against ALKWT, ALKL1196M, ALKG1202R and ROS1WT, respectively. Subsequently, western blot assay showed that compound 16 significantly suppressed ALK and its downstream protein expression in a dose-dependent manner. Alternatively, the Hoechst 33258 and AO/EB staining assays illustrated that compound 16 could induce H2228 cell apoptosis. Ultimately, the binding models of compound 16 with ALKWT, ALKG1202R as well as ROS1 clearly presented the essential interactions within the active site. Together, compound 16 was validated as a promising ALK/ROS1 dual inhibitor for ALKG1202R mutation correlated tumors.