Chemoproteomics-Enabled Covalent Ligand Screening Reveals a Thioredoxin-Caspase 3 Interaction Disruptor That Impairs Breast Cancer Pathogenicity
Chemoproteomics-Enabled Covalent Ligand Screening Reveals a Thioredoxin-Caspase 3 Interaction Disruptor That Impairs Breast Cancer Pathogenicity
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DOI：
10.1021/acschembio.7b00711
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发表时间：
2017-10-01
影响因子：
4
通讯作者：
Nomura, Daniel K.
中科院分区：
文献类型：
--
作者：
Anderson, Kimberly E.;To, Milton;Nomura, Daniel K.
发现共价配体是开发针对治疗靶点的小分子效应器的一种很有前途的策略。最近的研究表明，二氯三嗪是一种很有前途的活性支架，它优先与赖氨酸发生反应。在这里，我们已经合成了一系列基于二氯三嗪的共价配体，并对这个文库进行了筛选，以揭示影响三阴性乳腺癌细胞存活的小分子。在从这个筛选KEA1-97中鉴定出一个铅点击后，我们使用基于活性的蛋白质图谱(ABPP)化学蛋白质组学平台来鉴定该化合物针对硫氧还蛋白的赖氨酸72-一个先前被证明在与caspase 3的蛋白质相互作用中重要的位置，以抑制caspase 3的活性和抑制细胞凋亡。我们发现KEA1-97在不影响硫氧还蛋白活性的情况下，破坏硫氧还蛋白与caspase3的相互作用，激活caspase，并诱导细胞凋亡。此外，KEA1-97还损害了体内乳腺癌移植瘤的生长。我们的研究展示了如何将共价配体的筛选与ABPP平台结合起来，以识别独特的抗癌先导和目标对。
Covalent ligand discovery is a promising strategy to develop small-molecule effectors against therapeutic targets. Recent studies have shown that dichlorotriazines are promising reactive scaffolds that preferentially react with lysines. Here, we have synthesized a series of dichlorotriazine-based covalent ligands and have screened this library to reveal small molecules that impair triple-negative breast cancer cell survival. Upon identifying a lead hit from this screen KEA1-97, we used activity-based protein profiling (ABPP)-based chemoproteomic platforms to identify that this compound targets lysine 72 of thioredoxin-a site previously shown to be important in protein interactions with caspase 3 to inhibit caspase 3 activity and suppress apoptosis. We show that KEA1-97 disrupts the interaction of thioredoxin with caspase 3, activates caspases, and induces apoptosis without affecting thioredoxin activity. Moreover, KEA1-97 impairs in vivo breast tumor xenograft growth. Our study showcases how the screening of covalent ligands can be coupled with ABPP platforms to identify unique anticancer lead and target pairs.