RIPK3 inhibitor-AZD5423 alleviates acute kidney injury by inhibiting necroptosis and inflammation.

Abstract

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, Arg37 might 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 Lys50 and 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.

• Authors: Chuan-Hui Xu, Jia-Nan Wang, Xiao-Guo Suo, Ming-Lu Ji, Xiao-Yan He, Xin Chen, Sai Zhu, Yuan He, Shuai-Shuai Xie, Chao Li, Ze-Hui Dong, Ying Chen, Wei-Jian Ni, Xiao-Wen Feng, Ming-Ming Liu, Juan Jin, Zeng Li, Xiao-Ming Meng
• Journal: International immunopharmacology (Int Immunopharmacol) Vol. 112 Pg. 109262 (Nov 2022) ISSN: 1878-1705 [Electronic] Netherlands
• PMID: 36166972 (Publication Type: Journal Article)
• Copyright: Copyright © 2022 Elsevier B.V. All rights reserved.

Chemical References

• 2,2,2-trifluoro-N-(1-((1-(4-fluorophenyl)-1H-indazol-5-yl)oxy)-1-(3-methoxyphenyl)-2-propanyl)acetamide
• Cisplatin
• Tumor Necrosis Factor-alpha
• Receptor-Interacting Protein Serine-Threonine Kinases
• Amino Acids
• Ripk3 protein, mouse

Topics

• Mice
• Animals
• Necroptosis
• Cisplatin (therapeutic use)
• Tumor Necrosis Factor-alpha (metabolism)
• Mice, Inbred C57BL
• Receptor-Interacting Protein Serine-Threonine Kinases (metabolism)
• Acute Kidney Injury (chemically induced)
• Inflammation (metabolism)
• Amino Acids