Aiming to explore novel anaplastic lymphoma kinase (ALK) and proto-oncogene tyrosine-protein kinase ROS (ROS1) dual inhibitors to overcome crizotinib-resistant mutants, two series of 2,4-diarylaminopyrimidine (DAAP) analogues bearing thiazole or 1,2,3-triazole moieties were designed and synthesized based upon the cocrystal structure of ceritinib with ALK^WT (PDB 4MKC) as well as the binding model of ceritinib with ALK^G1202R. The cellular and enzymatic assays validated 34c (WY-135) as a promising ALK (IC₅₀ = 1.4 nM) and ROS1 (IC₅₀ = 1.1 nM) dual inhibitor superior to crizotinib and ceritinib. 34c showed significantly inhibitory activities on ALK-dependent cell lines KARPAS299 (IC₅₀ = 21 nM) and H2228 (IC₅₀ = 95 nM) as well as ROS1-positive cell line HCC78 (IC₅₀ = 40 nM). In particular, 34c was potent against a variety of frequently observed crizotinib-resistant mutants, particularly the L1196M mutant (IC₅₀ = 3.1 nM) identified as the “gatekeeper” mutation and the G1202R mutant (IC₅₀ = 8.7 nM) which conferred resistance to all clinical stage ALK inhibitors. Furthermore, 34c was capable of inducing cell apoptosis and strongly inhibiting cellular ALK and ROS1 activity. In addition, the binding models of 34c with ALK^WT, ALK^L1196M and ALK^G1202R provided structural bases for SARs observations.

旨在探索新型的间变性淋巴瘤激酶（ALK）和原癌基因酪氨酸蛋白激酶ROS（ROS1）双重抑制剂，以克服耐克唑替尼的突变体，两个系列的带有噻唑或1,2的2,4-二芳基氨基嘧啶（DAAP）类似物， 3-三唑基部分被设计和基于ceritinib与ALK共晶体结构合成^WT（PDB 4MKC）以及ceritinib与ALK结合模型^G1202R。细胞和酶促测定证实34c（WY-135）是 优于crizotinib和ceritinib的有前景的ALK（IC ₅₀  = 1.4 nM）和ROS1（IC ₅₀ = 1.1 nM）双重抑制剂。34c对ALK依赖的细胞系KARPAS299（IC ₅₀  = 21 nM）和H2228（IC ₅₀  = 95 nM）以及ROS1阳性细胞系HCC78（IC ₅₀  = 40 nM）表现出显着的抑制活性。特别是，34c对多种经常观察到的耐克唑替尼耐药的突变体有效，特别是被 鉴定为“关守”突变的L1196M突变体（IC ₅₀ = 3.1 nM）和 赋予抗药性的G1202R突变体（IC ₅₀ = 8.7 nM）。所有临床阶段的ALK抑制剂。此外，34c能够诱导细胞凋亡并强烈抑制细胞ALK和ROS1活性。此外，34c与ALK的绑定模型^WT，ALK ^L1196M和ALK ^G1202R为SAR观测提供了结构基础。