Specific, irreversible inactivation of the epidermal growth factor receptor and erbB2, by a new class of tyrosine kinase inhibitor
Specific, irreversible inactivation of the epidermal growth factor receptor and erbB2, by a new class of tyrosine kinase inhibitor
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DOI：
10.1073/pnas.95.20.12022
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发表时间：
1998-09-29
影响因子：
11.1
通讯作者：
Dobrusin, EM
中科院分区：
文献类型：
--
作者：
Fry, DW;Bridges, AJ;Dobrusin, EM
本发明公开了一类高亲和力抑制剂，它通过对存在于ATP结合口袋中的半胱氨酸残基进行特异的共价修饰，选择性地靶向并不可逆地灭活表皮生长因子受体酪氨酸激酶。利用MS、分子模拟、定点突变和活细胞中的C-14标记进行的一系列实验明确地证明，这些化合物以1：1的化学计量比和烷基化半胱氨酸-773选择性地与表皮生长因子受体的催化域结合。虽然这些化合物在溶液中基本上不具反应性，但当它们结合在ATP口袋中时，它们会受到这种特殊氨基酸的快速亲核攻击。这些抑制剂中丙烯酰胺基团相对于Cys-773的分子取向和定位完全支持这些结果，这些结果是通过在同源建立的ATP位点的分子模型中进行对接实验确定的。也有证据表明，这些化合物与erbB2以类似的方式相互作用，但对本研究中测试的其他受体酪氨酸激酶或细胞内酪氨酸激酶没有活性。最后，6-丙烯酰胺-4-苯胺喹唑啉与一种同样有效但可逆的类似物之间的直接比较表明，这种不可逆抑制剂在人表皮样癌移植瘤模型中具有明显的体内抗肿瘤活性，在治疗活性剂量下没有明显的毒性。这种化合物的活性图谱是一代酪氨酸激酶抑制剂的典型代表，在治疗增生性疾病方面具有很大的治疗意义。
A class of high-affinity inhibitors is disclosed that selectively target and irreversibly inactivate the epidermal growth factor receptor tyrosine kinase through specific, covalent modification of a cysteine residue present in the ATP binding pocket. A series of experiments employing MS, molecular modeling, site directed mutagenesis, and C-14-labeling studies in viable cells unequivocally demonstrate that these compounds selectively bind to the catalytic domain of the epidermal growth factor receptor with a 1:1 stoichiometry and alkylate Cys-773. While the compounds are essentially nonreactive in solution, they are subject to rapid nucleophilic attack by this particular amino acid when bound in the ATP pocket. The molecular orientation and positioning of the acrylamide group in these inhibitors in relation to Cys-773 entirely support these results as determined from docking experiments in a homology-built molecular model of the ATP site. Evidence is also presented to indicate that the compounds interact in an analogous fashion with erbB2 but have no activity against the other receptor tyrosine kinases or intracellular tyrosine kinases that were tested in this study. Finally, a direct comparison between 6-acrylamido-4-anilinoquinazoline and an equally potent but reversible analog shows that the irreversible inhibitor has far superior in vivo antitumor activity in a human epidermoid carcinoma xenograft model with no overt toxicity at therapeutically active doses. The activity profile for this compound is prototypical of a generation of tyrosine kinase inhibitors with great promise for therapeutic significance in the treatment of proliferative disease.