ALK及ROS1融合蛋白是非小细胞肺癌等肿瘤个性化治疗的重要分子靶点，但激酶结构域突变引起的耐药使现有药物无效。本项目旨在探索“基于激酶突变位点的立体构象构建抗突变耐药的ALK/ROS1选择性抑制剂”的可行性。以前期发现的对野生型及突变型ALK/ROS1均有较强抑制活性的2-氨基嘧啶衍生物ZXN-17为先导化合物，针对G1202R ALK/G2032R ROS1突变位点，本项目拟通过化合物与突变蛋白晶体结构的分子对接、动力学模拟手段，采用"Mix-n-match"杂合策略，构建满足突变蛋白构象要求的化合物库，合成8类目标分子，通过多水平的活性筛选及安全性评价，优选出1-3个先导化合物；开展先导物与靶标的共晶研究，结合对靶标下游信号通路的研究阐明其作用机制，解决①如何设计对突变靶标有效的分子；②如何验证其活性与机制两个关键问题，为抗突变耐药的ALK/ROS1选择性抑制剂的设计提供科学依据。

ALK- and ROS1-oncogene fusion proteins are effective molecular targets for “personalized medicine” in non-small celll lung cancers, but the mutations in kinase domain of ALK-and ROS1-oncogene fusion proteins are responsible for ALK inhibitor resistance in these cancers. It is a medicinal chemistry campaign aimed at improving the feasibility of built selected ALK/ROS1 dual inhibitors against crizotinib-resistant mutants based on the structural conformation of mutation point in kinase domain. In our previous study, we found a 2-aminopyrimidine compound ZXN-17 which showed strong ALK and ROS1 inhibitory activity. Encouraged by previous results, aiming to G1202R ALK/G2032R ROS1 mutation points,this project will develop multiplex inhibitors of ALK- and ROS1 based on their mutation conformation by using of molecular Docking and molecular dynamics simulation. The design strategies of "mix-n-match" and "hybridization" will be adopted with ZXN-17 as lead compound. Eight kinds of molecules will be synthesized. In order to evaluate the potency and selectivity of the title compounds, various biological assays and primary safety evaluation will be carried out. This project will identify 1-3 lead compound with a "dual" ALK/ROS1inhibition profile. The mechanisms of action of lead compound will be illuminated by means of co-crystal research between lead with target protein combined with the research on the downstream signaling pathways of ALK and ROS1 targets. The project would resovle two questions. One is how to design the effective molecules against the mutated ALK and ROS1; the other is how to identify the viability, activity and mechanisms of the title compounds. As a result, this project may provide the scientific basis to the design of novel ALK/ROS1 selective dual inhibitors against crizotinib resistant mutants.