An inhibitor of Nrf2
ML385 is a specific nuclear factor erythroid 2-related factor 2 (NRF2) inhibitor. ML385 binds to NRF2 and inhibits its downstream target gene expression as a probe molecule. Specifically, ML385 binds to the Neh1, the Cap ‘N’ Collar Basic Leucine Zipper (CNC-bZIP) domain of NRF2, and interferes with the binding of the V-Maf Avian Musculoaponeurotic Fibrosarcoma Oncogene Homolog G (MAFG)-NRF2 protein complex to regulatory DNA binding sequences. ML385 shows specificity and selectivity for NSCLC cells with KEAP1 mutation leading to gain of NRF2 function.[1][2]
In vitro study demonstrated that ML385 potentially inhibits NRF2 through direct interaction. The NRF2 signaling was decreased in a time-dependent manner and the maximum decline was at 72 h. A reduction in NRF2 mRNA levels was also observed. In addition, ML385 caused global inhibition of NRF2 signaling in lung cancer cells with KEAP1 mutations, and other target genes. ML385 treatment also significantly attenuated NQO1 enzyme activity and reduced GSH levels along with cellular antioxidant capacity. [2]
In vivo study of ML385 indicated that it inhibits NRF2 and showed promising anti-tumor activity. ML385 in combination with carboplatin showed a significant reduction in tumor growth compared to vehicle. Although the treatment with a single agent (either ML385 or carboplatin) led to a reduction in tumor growth, the magnitude of these effects was variable between cell lines and did not reach statistical significance. Moreover, tumor samples treated with ML385 showed a significant reduction in NRF2 protein level and its downstream target genes. Besides, the addition of ML385 decreased anisotropy in a dose-dependent manner, with an IC50 of 1.9 μM, suggesting that the NRF2-MAFG protein complex was dissociated from fluorescein-labeled ARE-DNA. [2]
ML385是一种特定的核因子红细胞2相关因子2（NRF2）抑制剂。它作为探针分子结合到NRF2并抑制其下游靶基因表达。具体来说，ML385结合到NRF2的CNC-bZIP域中的Neh1，并干扰V-Maf鸟类肌腱性纤维肉瘤癌基因同源物G（MAFG）- NRF2蛋白复合物与调节DNA结合序列的结合。 ML385对于KEAP1突变导致增益NRF2功能的非小细胞肺癌细胞具有特异性和选择性。 [1] [2]
体外研究表明，ML385可能通过直接作用来抑制NRF2。 NRF2信号传导随时间的推移而减少，最大下降在72小时时出现。还观察到了NRF2 mRNA水平的降低。此外，ML385引起肺癌细胞中KEAP1突变和其他靶基因的全局抑制。 ML385治疗还显著减弱了NQO1酶活性，并降低了GSH水平以及细胞抗氧化能力。 [2]
ML385的体内研究表明，它可以抑制NRF2，并显示出有希望的抗肿瘤活性。与仅使用载体相比，ML385与卡铂联合使用可显著减少肿瘤生长。虽然单一药物治疗（ML385或卡铂）导致了肿瘤生长的减少，但这些效果在细胞系之间变化很大，并且没有达到统计学意义水平。此外，经过ML385处理的肿瘤样本显示出NRF2蛋白水平及其下游靶基因显著降低。此外，添加ML385以剂量依赖方式降低各向异性，在IC50为1.9μM时提示NRF2-MAFG蛋白复合物从荧光素标记的ARE-DNA中解离。[2]
参考文献:
[1]. Xian P, et al. Mesenchymal stem cell-derived exosomes as a nanotherapeutic agent for amelioration of inflammation-induced astrocyte alterations in mice. Theranostics. 2019 Aug 14;9(20):5956-5975.
[2]. Singh A, et al. Small Molecule Inhibitor of NRF2 Selectively Intervenes Therapeutic Resistance in KEAP1-Deficient NSCLC Tumors. ACS Chem Biol. 2016 Nov 18;11(11):3214-3225.
| Cell experiment [1]: | |
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Cell lines |
A549 cells |
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Preparation Method |
A549 cells were transfected with a firefly luciferase reporter (Fluc) construct driven by a minimal TATA promoter with upstream NRF2-specific antioxidant response element (ARE) enhancer sequence from human NQO1 promoter ARE and clones stably expressing ARE-FLuc7 were screened and validated. |
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Reaction Conditions |
Cells were cultured in the presence of ML385 (5 μM) for 48h and 72 h and measured the changes in the expression levels of NRF2 and its target genes. |
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Applications |
ML385 treatment to A549 cells demonstrated a reduction in glutathione synthesis and recycling enzymes, members of the thioredoxin family, and glucose metabolism-related genes with a time-dependent manner. ML385 treatment significantly attenuated NQO1 enzyme activity and reduced GSH levels along with cellular antioxidant capacity. |
| Animal experiment [2]: | |
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Animal models |
1-8-week-old C57B/6 male mice |
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Preparation Method |
Mice received a daily intraperitoneal injection of ML385 (30 mg/kg) dissolved in PBS with 5% Dimethyl Sulfoxide (DMSO) for 7 d. |
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Dosage form |
30 mg/kg |
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Applications |
ML385 could inhibit Nrf2 and used to explore the mechanism of Nrf2-NF-κB signaling pathway involved in the inhibition of astrocyte activation. Mice treated with ML385 had a significant decrease in Keap1, HO-1, Nrf2, and p-P65/P-65 (P < 0.05) expression compared to Sham or Vehicle control groups. |
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参考文献: [1]. Singh A, et al. Small Molecule Inhibitor of NRF2 Selectively Intervenes Therapeutic Resistance in KEAP1-Deficient NSCLC Tumors. ACS Chem Biol. 2016 Nov 18;11(11):3214-3225. [2]. Xian P, et al. Mesenchymal stem cell-derived exosomes as a nanotherapeutic agent for amelioration of inflammation-induced astrocyte alterations in mice. Theranostics. 2019 Aug 14;9(20):5956-5975. |
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