Development of an enzyme-linked immunosorbent assay for Keap1-Nrf2 interaction inhibitors identification

• Published in: Redox biology 2020-07, Vol.34, p.101573-101573, Article 101573
• Main Authors: Wang, Yan, Xiao, Chu-Ying, Lin, Huang-Quan, Hu, Jian-Shu, Ip, Tsz-Ming, Chi-Cheong Wan, David
• Format: Article
• Language: English
• Subjects: ELISA; FDA approved Drugs; Keap1-Nrf2 interaction inhibitor; Short Communication
• ISSN: 2213-2317; 2213-2317
• DOI: 10.1016/j.redox.2020.101573

Development of Keap1–Nrf2 interaction inhibitors is a promising strategy for the discovery of therapeutic agents against oxidative stress-mediated diseases. Two motifs of Nrf2, ETGE and DLG motif, are responsible for Keap1-Nrf2 binding. Previously, ETGE peptide or ETGE-derived peptide-based approaches were used to detect Keap1-Nrf2 interaction; however, these approaches are not able to monitor Keap1-DLG motif binding. We first report here a novel Enzyme-linked Immunosorbent Assay (ELISA) approach to detect the protein-protein interaction of full length Keap1 and Nrf2. In our assay, the test compounds can target either ETGE or DLG binding site, therefore facilitating the exploration of diverse Keap1-Nrf2 inhibitors. Three FDA-approved drugs, zafirlukast, dutasteride and ketoconazole, were found to inhibit the Keap1-Nrf2 interaction with IC50 of 5.87, 2.81 and 1.67 μM, respectively. Additionally, these three drugs also activated Nrf2 pathway in neuroblasts and lipopolysaccharide (LPS)-challenged mice. The results presented here indicate that the ELISA approach has the capacity to identify Keap1-Nrf2 inhibitors. [Display omitted] •We established a novel Enzyme-linked Immunosorbent Assay for Keap1-Nrf2 interaction inhibitors identification.•This ELISA is the first approach to detect the protein-protein interaction of full length Keap1 and Nrf2.•In this assay, the test compounds can target either ETGE or DLG binding site, therefore facilitating the exploration of diverse Keap1-Nrf2 inhibitors.•Zafirlukast, dutasteride and ketoconazole have been identified as inhibitors of the Keap1-Nrf2 interaction.