Selective and reversible modification of kinase cysteines with chlorofluoroacetamides
Selective and reversible modification of kinase cysteines with chlorofluoroacetamides
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DOI：
10.1038/s41589-018-0204-3
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发表时间：
2019-03-01
影响因子：
14.8
通讯作者：
Ojida, Akio
中科院分区：
文献类型：
--
作者：
Shindo, Naoya;Fuchida, Hirokazu;Ojida, Akio
用小分子不可逆地抑制疾病相关蛋白是实现增加和持续药理效力的有效方法。在这里，我们介绍α-氯氟乙酰胺（CFA）作为靶向共价抑制剂（TCI）的新型弹头。尽管内在反应性较弱，但添加 CFA 的喹唑啉对表皮生长因子受体 (EGFR) 的 Cys797 显示出高反应性。在细胞中，CFA-喹唑啉在较宽的浓度范围（0.1-10 μM）内对 EGFR 表现出比相应的 Michael 受体更高的靶特异性。 CFA 衍生物的半胱氨酸加合物易于水解并可逆地产生完整的硫醇，但在 EGFR 的溶剂隔离的 ATP 结合袋中保持稳定。这种环境依赖性水解可能会减少基于 CFA 的药物对蛋白质的脱靶修饰。口服 CFA 喹唑啉 NS-062 显着抑制小鼠异种移植模型中的肿瘤生长。此外，附加 CFA 的吡唑并嘧啶不可逆地抑制布鲁顿酪氨酸激酶，具有更高的靶点特异性。这些结果证明了 CFA 作为 TCI 新型弹头的实用性。
Irreversible inhibition of disease-associated proteins with small molecules is a powerful approach for achieving increased and sustained pharmacological potency. Here, we introduce alpha-chlorofluoroacetamide (CFA) as a novel warhead of targeted covalent inhibitor (TCI). Despite weak intrinsic reactivity, CFA-appended quinazoline showed high reactivity toward Cys797 of epidermal growth factor receptor (EGFR). In cells, CFA-quinazoline showed higher target specificity for EGFR than the corresponding Michael acceptors in a wide concentration range (0.1-10 mu M). The cysteine adduct of the CFA derivative was susceptible to hydrolysis and reversibly yielded intact thiol but was stable in solvent-sequestered ATP-binding pocket of EGFR. This environment-dependent hydrolysis can potentially reduce off-target protein modification by CFA-based drugs. Oral administration of CFA quinazoline NS-062 significantly suppressed tumor growth in a mouse xenograft model. Further, CFA-appended pyrazolopyrimidine irreversibly inhibited Bruton's tyrosine kinase with higher target specificity. These results demonstrate the utility of CFA as a new class warheads for TCI.