Institute of Developmental Genetics , Helmholtz Zentrum München , 85764 Neuherberg , Germany.
Rudolf Virchow Center for Experimental Biomedicine , University of Würzburg , 97080 Würzburg , Germany.
J Med Chem. 2019 Jan 10;62(1):266-275. doi: 10.1021/acs.jmedchem.8b00315. Epub 2018 May 2.
Glutathione peroxidase 4 (GPX4) is essential for cell membrane repair, inflammation suppression, and ferroptosis inhibition. GPX4 upregulation provides unique drug discovery opportunities for inflammation and ferroptosis-related diseases. However, rational design of protein activators is challenging. Until now, no compound has been reported to activate the enzyme activity of GPX4. Here, we identified a potential allosteric site in GPX4 and successfully found eight GPX4 activators using a novel computational strategy and experimental studies. Compound 1 from the virtual screen increased GPX4 activity, suppressed ferroptosis, reduced pro-inflammatory lipid mediator production, and inhibited NF-κB pathway activation. Further chemical synthesis and structure-activity relationship studies revealed seven more activators. The strongest compound, 1d4, increased GPX4 activity to 150% at 20 μM in the cell-free assay and 61 μM in cell extracts. Therefore, we demonstrated that GPX4 can be directly activated using chemical compounds to suppress ferroptosis and inflammation. Meanwhile, the discovery of GPX4 activators verified the possibility of rational design of allosteric activators.
谷胱甘肽过氧化物酶 4（GPX4）对于细胞膜修复、炎症抑制和铁死亡抑制至关重要。GPX4 的上调为炎症和铁死亡相关疾病的药物发现提供了独特的机会。然而，蛋白质激活剂的合理设计具有挑战性。到目前为止，还没有报道称有化合物能够激活 GPX4 的酶活性。在这里，我们鉴定了 GPX4 中的一个潜在别构位点，并成功使用一种新的计算策略和实验研究发现了八种 GPX4 激活剂。虚拟筛选出的化合物 1 增加了 GPX4 的活性，抑制了铁死亡，减少了促炎脂质介质的产生，并抑制了 NF-κB 通路的激活。进一步的化学合成和构效关系研究揭示了另外七种激活剂。最强的化合物 1d4 在无细胞测定中在 20μM 时将 GPX4 活性提高到 150%，在细胞提取物中提高到 61μM。因此，我们证明了可以使用化学化合物直接激活 GPX4 来抑制铁死亡和炎症。同时，GPX4 激活剂的发现验证了合理设计别构激活剂的可能性。
