Optimization of the C2 substituents on the 1,4-bis(arylsulfonamido)naphthalene-N,N'-diacetic acid scaffold for better inhibition of Keap1-Nrf2 protein-protein interaction.
Optimization of the C2 substituents on the 1,4-bis(arylsulfonamido)naphthalene-N,N'-diacetic acid scaffold for better inhibition of Keap1-Nrf2 protein-protein interaction.
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DOI：
10.1016/j.ejmech.2023.115302
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发表时间：
2023-03
影响因子：
6.7
通讯作者：
Dhulfiqar Ali Abed;Ahmed R. Ali;Sumi Lee;Mai-Uyen Nguyen;M. Verzi;Longqin Hu
中科院分区：
文献类型：
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作者：
Dhulfiqar Ali Abed;Ahmed R. Ali;Sumi Lee;Mai-Uyen Nguyen;M. Verzi;Longqin Hu
直接抑制Kelch样ECH相关蛋白1（Keap 1）和核因子红细胞2相关因子2（Nrf 2）之间的蛋白质-蛋白质相互作用（PPI），可减少Nrf 2的泛素化和随后的降解，导致Nrf 2在胞质溶胶中的积累和Nrf 2的核转位。一旦进入细胞核，Nrf 2结合并激活参与氧化还原稳态和解毒的抗氧化反应元件（ARE）基因的表达。本文报道了一系列具有不同C2取代基的1，4-双（芳基磺酰氨基）萘-N，N′-二乙酸类似物，以探索中心萘核这一位置的构效关系。首先用荧光偏振（FP）测定筛选Keap 1结合活性，然后使用更灵敏的时间分辨荧光能量转移（TR-FRET）测定进一步评价更有效的化合物。发现具有C2-邻苯二甲酰亚氨基丙基的化合物24 a在该系列中是最有效的，在TR-FRET测定中显示2.5 nM的IC 50，Ki值在亚纳摩尔范围内。我们的对接研究表明，C2-邻苯二甲酰亚胺丙基在化合物24 a提供了一个额外的氢键与关键残基Arg 415的相互作用，这可能是负责所观察到的结合亲和力的提高。此外，化合物12 b、15和24 a显示激活NCM 460 D细胞中的Nrf 2信号传导途径，导致GSTM 3、HMOX 1和NQO 1的mRNA水平在100 μM下与溶媒对照相比升高2.4-11.7倍。
Direct inhibition of the protein-protein interaction (PPI) between Kelch-like ECH-associated protein 1 (Keap1) and nuclear factor erythroid 2-related factor 2 (Nrf2) reduces the ubiquitination and subsequent degradation of Nrf2, leading to Nrf2 accumulation in the cytosol and the nuclear translocation of Nrf2. Once inside the nucleus, Nrf2 binds to and activates the expression of antioxidant response element (ARE) genes involved in redox homeostasis and detoxification. Herein, we report a series of 1,4-bis(arylsulfonamido)naphthalene-N,N′-diacetic acid analogs with varying C2 substituents to explore the structure-activity relationships at this position of the central naphthalene core. The Keap1-binding activities were first screened with a fluorescence polarization (FP) assay followed by further evaluation of the more potent compounds using a more sensitive time-resolved fluorescence energy transfer (TR-FRET) assay. It was found that compound24awith C2-phthalimidopropyl group was the most potent in this series showing an IC50of 2.5 nM in the TR-FRET assay with a Kivalue in the subnanomolar range. Our docking study indicated that the C2-phthalimidopropyl group in compound24aprovided an extra hydrogen bonding interaction with the key residue Arg415 that may be responsible for the observed boost in binding affinity. In addition, compounds12b,15, and24awere shown to activate the Nrf2 signaling pathway in NCM460D cells resulting in elevated mRNA levels ofGSTM3,HMOX1andNQO1by 2.4–11.7 fold at 100 μM as compared to the vehicle control.