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目的：用计算机辅助设计法寻找 HER2/neu受体酪氨酸激酶小分子抑制剂。方法：用 MODERLAR软件模拟出 HER2/neu和 EGFR受体酪氨酸激酶的三维 (three- dimensional,3D)空间结构；根据 HER2/neu酪氨酸激酶 ATP结合区的空间结构，搜索数据库，找出候选化合物；用 Western blot方法检测化合物对 HER2/neu受体酪氨酸激酶磷酸化的影响； MTT法检测细胞增殖抑制作用。结果：比较 HER2/neu和 EGFR（靶蛋白）与胰岛素受体酪氨酸激酶、 FGFR1受体酪氨酸激酶和 Src酪氨酸激酶（模板蛋白）的氨基酸序列发现有 35％～ 41％的相同和 52％～ 55％的相似，用 MODELLER程序模拟出 HER2/neu和 EGFR激酶区域的 3D结构。根据 HER2/neu受体酪氨酸激酶结构模型， 3D数据库的搜索，获得潜在 HER2/neu酪氨酸激酶抑制剂化合物，经筛选、优化发现 ST2325对 HER2/neu磷酸化有明显的抑制作用，其 IC50值为 6.6μ mol/L，且抑制作用是可逆的，对 EGFR受体磷酸化没有抑制作用。 ST2325对 HER2/neu受体表达没有影响。 ST2325对 HER2/neu过表达的肿瘤细胞 MDA- MB- 453 m1的抑制作用更强，而对 EGFR过表达的肿瘤细胞 MDA- MB- 468抑制作用相对较弱，其 IC50值分别为 29.05μ mol/L和 60.40μ mol/L。结论：基于 HER2/neu的结构设计，筛选?
Objective: To search for HER2/neu small molecule tyrosine kinase inhibitors using computer-aided design. METHODS: Three-dimensional (3D) spatial structures of HER2/neu and EGFR receptor tyrosine kinases were modeled using MODELARLAR software; the database was searched based on the spatial structure of the HER2/neu tyrosine kinase ATP binding region. Candidate compounds; The effect of compounds on the phosphorylation of HER2/neu receptor tyrosine kinases was detected by Western blot; The inhibition of cell proliferation was measured by MTT assay. Results: The amino acid sequences of HER2/neu and EGFR (target proteins) and insulin receptor tyrosine kinase, FGFR1 receptor tyrosine kinase and Src tyrosine kinase (template protein) were found to be 35% to 41% identical. Similar to 52% to 55%, the 3D structure of the HER2/neu and EGFR kinase domains was modeled using the MODELLER program. According to the structural model of HER2/neu receptor tyrosine kinase and 3D database search, potential HER2/neu tyrosine kinase inhibitor compounds were obtained. After screening and optimization, ST2325 was found to have obvious inhibitory effect on HER2/neu phosphorylation. The IC50 value was 6.6μmol/L, and the inhibitory effect was reversible. There was no inhibitory effect on EGFR receptor phosphorylation. ST2325 had no effect on HER2/neu receptor expression. ST2325 had stronger inhibition of HER2/neu overexpressing tumor cell MDA-MB-453 m1, while the inhibitory effect of EGFR over-expressing tumor cell MDA-MB-468 was relatively weak. Its IC50 value was 29.05 μ mol/ L and 60.40 μmol/L. Conclusion: Based on the structural design of HER2/neu, screening?