Aiming to develop promising ALK inhibitors, two series of N²-(2-alkyoxy-6-aliphatic aminopyridin-3-yl)-2,4-diaminepyrimidine derivatives (22a-x and 23a-d) were designed according to scaffold hopping and bioisosterism principles. All compounds were efficiently synthesized by concise reactions and anti-proliferative activities on ALK-addicted H2228, Karpas299 cells and EGFR-expressive A549 cell were evaluated by MTT assay. Several compounds exhibited potential cytotoxic activities with IC₅₀ values below 0.10 μM. Five compounds (22g, 22h, 22l, 22s and 23a) were selected for further enzymatic determination, resulting in the discovery of 22l against ALK and ALK^L1196M with IC₅₀ values of 2.1 nM and 3.8 nM. Particularly, western blot and cell apoptosis assays identified 22l as a promising ALK inhibitor, which was capable of obviously inhibiting cellular ALK activity and inducing cell apoptosis. Eventually, molecular docking modes of 22l with ALK confirmed structural basis in accordance with the SARs analysis.

为了开发有前景的ALK抑制剂，根据支架跳跃和串联效应，设计了两个系列的N ^2-（2-烷氧基-6-脂族氨基吡啶-3-基）-2,4-二胺嘧啶衍生物（22a-x和23a-d）。生物立体异构原则。通过简明反应有效地合成了所有化合物，并通过MTT分析评估了对ALK成瘾的H2228，Karpas299细胞和表达EGFR的A549细胞的抗增殖活性。几种化合物表现出潜在的细胞毒性活性，IC ₅₀值低于0.10μM。五种化合物（22g，22h，22l，22s和23a）被选择用于进一步酶法测定，结果的发现22升对抗ALK和ALK ^L1196M带IC ₅₀ 2.1纳米和3.8纳米的值。特别地，蛋白质印迹和细胞凋亡测定法鉴定出22l是有前途的ALK抑制剂，其能够明显抑制细胞ALK活性并诱导细胞凋亡。最终，根据SAR分析，具有ALK的22l分子对接模式确定了结构基础。