PO.ET02.12 · 实验与分子治疗
发现一种具有抗肿瘤活性的新型共价异双功能硫氧还蛋白还原酶抑制剂
Discovery of a novel, covalent and heterobifunctional thioredoxin reductase inhibitor with antitumor activity
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
活性氧（ROS）在细胞信号传导和稳态中发挥核心作用。然而，ROS水平失衡会导致包括癌症在内的多种病理状态。肿瘤细胞常上调抗氧化系统，如硫氧还蛋白还原酶（TrxR）和谷胱甘肽（GSH）通路，以缓冲氧化应激。异常代谢和ROS的发现，作为肿瘤一个可靶向的标志性易感靶点，引发了极大的兴奋和期望。遗憾的是，尽管学术界和产业界付出了巨大努力，这一期望仍未实现。在此，我们报道利用去泛素化酶靶向嵌合体（DUBTAC）技术，发现YS207，一种首创的共价异双功能TrxR1抑制剂。YS207由TrxR1募集部分通过一种新型化学连接子偶联去泛素化酶募集部分组成，从而稳定共价抑制的TrxR1，以深度生成治疗水平的ROS。YS207处理在体外和体内均强效抑制TrxR1酶活性（IC50分别为40nM和700nM）。机制上，YS207升高ROS水平，诱导脂质过氧化，并降低细胞内GSH。此外，我们通过透射电子显微镜（TEM）观察到线粒体的深度改变，包括肿胀、嵴丢失、电子密度增加和显著的线粒体自噬，均为氧化损伤的特征。用ROS清除剂N-乙酰半胱氨酸（NAC）进行的挽救实验证实，YS207诱导的细胞死亡是ROS依赖性的。在885种癌细胞系中进行的大规模PRISM分析显示，对YS207的敏感性与TrxR1依赖性、NADPH代谢和硒蛋白生物合成通路强烈相关。值得注意的是，TrxR1和GPX4的双敲除部分挽救了YS207诱导的细胞毒性，突显了它们在氧化还原调控中的协同作用。此外，我们证明了YS207在肺癌异种移植模型中具有可观的抗肿瘤生长抑制活性。综上所述，我们的结果表明YS207是一种强效的ROS调节剂，通过新型作用机制具有潜在的抗肿瘤活性。我们计划在对氧化还原失衡易感性增强的肿瘤中进一步开发这类分子。
查看英文原文 English abstract
Reactive oxygen species (ROS) play a central role in cellular signaling and homeostasis. However, imbalances in ROS levels contribute to various pathologies, including cancer. Tumor cells often upregulate antioxidant systems, such as the thioredoxin reductase (TrxR) and glutathione (GSH) pathways, to buffer against oxidative stress. The identification of aberrant metabolism and ROS has generated excitement and promise as a hallmark targetable vulnerability of tumors. Unfortunately, despite tremendous academic and industry efforts, this promise remains unfulfilled. Here, we report the discovery of YS207, a first-in-class covalent heterobifunctional inhibitor of TrxR1 using the deubiquitinase-targeting chimera (DUBTAC) technology. YS207 is composed of TrxR1 recruiting moiety coupled to deubiquitinase recruiting moiety via a novel chemical linker resulting in stabilization of covalently inhibited TrxR1 for profound generation of therapeutic levels of ROS. YS207 treatment potently inhibited TrxR1 enzymatic activity in vitro and in vivo (IC 50 of 40nM & 700nM respectively). Mechanistically, YS207 elevated ROS levels, induced lipid peroxidation, and decreased intracellular GSH. Additionally, we observed profound alterations of the mitochondria including swelling, loss of cristae, increased electron density and prominent mitophagy, all characteristic of oxidative damage by using transmission electron microscopy (TEM). Rescue experiments with the ROS scavenger N-acetyl cysteine (NAC) confirmed that YS207-induced cell death was ROS-dependent. Large-scale PRISM profiling across 885 cancer cell lines revealed that sensitivity to YS207 strongly correlates with TrxR1 dependency, NADPH metabolism, and selenoprotein biosynthesis pathways. Notably, dual knockout of TrxR1 and GPX4 partially rescued YS207-induced cytotoxicity, underscoring a cooperative role in redox regulation. Further, we showed promising anti-tumor growth inhibitory activity of YS207 in xenograft models of lung cancer. Taken together, our results demonstrate that YS207 is a potent ROS modulator with potential antitumor activities through novel mechanism of action. We aim to pursue further development of this class of molecules in tumors with enhanced vulnerability to redox imbalance.
利益披露 Disclosure
Y. Han, None..
N. Blazanin, None..
S. Kotagiri, None..
M. Qudratullah, None..
X. Liang, None..
Y. Lissanu, None.