RIPK3 inhibitor-AZD5423 alleviates acute kidney injury by inhibiting necroptosis and inflammation.
RIPK3 inhibitor-AZD5423 alleviates acute kidney injury by inhibiting necroptosis and inflammation.
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DOI：
10.1016/j.intimp.2022.109262
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发表时间：
2022-09
影响因子：
5.6
通讯作者：
Chuanjie Xu;Jia-nan Wang;Xiao-guo Suo;M. Ji;Xiao-yan He;Xin Chen;Sai Zhu;Yuan He;
中科院分区：
文献类型：
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作者：
Chuanjie Xu;Jia-nan Wang;Xiao-guo Suo;M. Ji;Xiao-yan He;Xin Chen;Sai Zhu;Yuan He;
急性肾损伤(AKI)是一种以肾小管上皮细胞炎症和程序性细胞死亡为特征的临床综合征，其定义为肾功能突然下降。坏死性下垂是一种受调控的细胞死亡形式，需要受体相互作用蛋白激酶3(RIPK3)的激活及其底物MLKL的磷酸化。RIPK3在急性肾损伤中起重要作用，因此开发其抑制剂被认为是预防和治疗AKI的有前途的策略之一。最近，我们根据RIPK3的三维结构，利用计算机辅助的混合虚拟筛选策略，发现了AZD5423作为一种新的有效的RIPK3抑制剂。我们的结果显示，AZD5423与先前发现的RIPK3抑制剂GSK872相比，显著抑制顺铂、缺氧/复氧(H/R)和肿瘤坏死因子-α刺激下RIPK3的激活和MLKL的磷酸化。重要的是，AZD5423对顺铂和缺血/再灌注(I/R)诱导的AKI小鼠模型具有有效的保护作用。细胞热位移实验和RIPK3基因敲除细胞的实验结果表明，AZD5423可以直接靶向RIPK3抑制RIPK3的活性。从机制上讲，AZD5423与RIPK3的对接表明RIPK3的Lys50、Arg313、Lys29、Arg37的激活区可能与AZD5423形成氢键。定点突变进一步表明，AZD5423通过与Lys50和Arg313的关键RIPK3氨基酸残基相互作用来降低伤害反应。综上所述，我们的研究表明，AZD5423可能是一种有效的RIPK3激酶抑制剂，并有望成为治疗AKI的临床候选药物。
Acute kidney injury (AKI) is a clinical syndrome that is defined as a sudden decline in renal function and characterized by inflammation and programmed cell death of renal tubular epithelial cells. Necroptosis is a form of regulated cell death that requires activation of receptor interacting protein kinase 3 (RIPK3) and its phosphorylation of the substrate MLKL. RIPK3 plays an important role in acute kidney injury, and hence developing its inhibitors is considered as one of the promising strategies aimed at prevention and treatment of AKI. Recently, we discovered AZD5423 as a novel potent RIPK3 inhibitor using a computer-aided hybrid virtual screening strategy according to three-dimensional structure of RIPK3. Our findings revealed that AZD5423 strongly inhibits activation of RIPK3, and MLKL phosphorylation upon cisplatin-, hypoxia/reoxygenation (H/R)- and TNF-α stimuli as compared with GSK872, which is a previously identified RIPK3 inhibitor. Importantly, AZD5423 exerts effective protection against cisplatin- and ischemia/reperfusion (I/R)-induced AKI mouse model. The results of cellular thermal shift assay and experiments in RIPK3 knockout cells indicated that AZD5423 could directly target RIPK3 to inhibit RIPK3 kinase activity. Mechanistically, the docking of AZD5423 and RIPK3 suggested that the kinase domain of RIPK3 for Lys50, Arg313, Lys29, Arg37might form hydrogen bonds with AZD5423. Site-directed mutagenesis further revealed that AZD5423 reduces injury response via interacting with the key RIPK3 amino acid residues of Lys50and Arg313. In conclusion, our study has demonstrated that AZD5423 may serve as a potent inhibitor of RIPK3 kinase and a promising clinical candidate for AKI treatment.