Acharya Reetuparna, Deb Pran Kishore, Pattanayak Shakti Prasad
Division of Advanced Pharmacology, Department of Pharmaceutical Sciences & Technology, Birla Institute of Technology, Mesra, 835215, Ranchi, India.
Division of Pharmaceutical Medicinal Chemistry, Department of Pharmaceutical Sciences & Technology, Birla Institute of Technology, Mesra, 835215, Ranchi, India.
Curr Med Chem. 2025 Jan 6. doi: 10.2174/0109298673356682241205044326.
The aim of this study is the evaluation of an Azomethine derivative, BCS2, for its antioxidant and anti-tumor activities against mammary carcinoma through the Nrf2- Keap1-HO-1 pathway.
The global prevalence of breast cancer is rising at an alarming rate. The facilitation of abnormal cell proliferation in mammary carcinoma occurs due to the disruption of signaling pathways that balance pro- and antioxidant status, thereby producing oxidative stress that disrupts genomic stability. Therefore, introducing a potent antioxidant molecule with antitumor activity is of paramount importance for treating breast cancer.
Synthesis, characterization, and in-vitro, in-vivo, and in-silico evaluation of an Azomethine derivative, BCS2, for its antioxidant and anti-tumor activities against chemical carcinogen- induced mammary carcinogenesis in Sprague-Dawley rats.
An azomethine derivative, 1-(4-nitrophenyl)-N-phenylmethanimine (BCS2), was synthesized and characterized based on its spectral data. The cytotoxic potential was observed on breast cancer cells, MCF-7, MDA-MB-231, and MDA-MB-468. The in vivo chemotherapeutic potential of BCS2 was established on 7,12-dimethylbenz(a)anthracene (DMBA) induced breast cancer in Sprague-Dawley (SD) rats. The effect of BCS2 on kelch-like ECH-associated protein- 1 (Keap1), Nrf2, heme oxygenase-1 (HO-1), mitogen-activated protein kinase (MAPK), and nuclear factor kappa-light-chain-enhancer of activated-B (NF-κB) was evaluated through ELISA and qPCR techniques. Furthermore, the binding potential and stability of BCS2 with Keap-1, HO-1, and MAPK were predicted using in silico molecular docking and dynamics studies. Additionally, drug-likeness properties of BCS2 were evaluated using in silico ADMET tools.
BCS2 showed remarkable cytotoxic activity on MCF-7 cells followed by MDA-MB- 231 and MDA-MB-468 cells having an IC50 of 2.368μM, 4.843μM and 6.472μM respectively, without affecting normal breast cells, MCF-10A. In the DMBA-induced animal model, BCS2 showed potent antitumor potential and showed protective action on endogenous-enzymatic and non-enzymatic antioxidants in cancer-bearing animals. Marked improvement in cellular architecture and ultrastructure of breast/tumor tissues excised from experimental animals was noted through histopathological and field emission scanning electron microscopy (FESEM) analyses. Significant upregulation of antioxidant proteins, Keap1 and HO-1, and downregulation of inflammatory proteins, MAPK, and NF-κB was observed after BCS2 treatment. The in silico computational studies predicted the potent binding of BCS2 with the active pockets of Keap1, HO-1, and MAPK proteins that validated the biological findings.
The study revealed BCS2's potent antioxidant and antitumor potential against mammary carcinoma through the Nrf2-Keap1-HO-1 signaling pathway.
本研究旨在评估一种甲亚胺衍生物BCS2通过Nrf2 - Keap1 - HO - 1途径对乳腺癌的抗氧化和抗肿瘤活性。
全球乳腺癌患病率正以惊人的速度上升。乳腺癌中异常细胞增殖的促进是由于平衡促氧化剂和抗氧化剂状态的信号通路受到破坏，从而产生破坏基因组稳定性的氧化应激。因此，引入具有抗肿瘤活性的强效抗氧化分子对于治疗乳腺癌至关重要。
合成、表征甲亚胺衍生物BCS2，并对其在体外、体内和计算机模拟方面针对化学致癌物诱导的Sprague - Dawley大鼠乳腺癌的抗氧化和抗肿瘤活性进行评估。
合成了一种甲亚胺衍生物1 - (4 - 硝基苯基)-N - 苯基亚甲胺（BCS2），并根据其光谱数据进行表征。观察了其对乳腺癌细胞MCF - 7、MDA - MB - 231和MDA - MB - 468的细胞毒性潜力。在Sprague - Dawley（SD）大鼠中建立了BCS2对7,12 - 二甲基苯并(a)蒽（DMBA）诱导的乳腺癌的体内化疗潜力。通过酶联免疫吸附测定（ELISA）和定量聚合酶链反应（qPCR）技术评估了BCS2对kelch样ECH相关蛋白1（Keap1）、核因子E2相关因子2（Nrf2）、血红素加氧酶 - 1（HO - 1）、丝裂原活化蛋白激酶（MAPK）和活化B细胞核因子κB轻链增强子（NF - κB）的影响。此外，使用计算机模拟分子对接和动力学研究预测了BCS2与Keap - 1、HO - 1和MAPK的结合潜力和稳定性。另外，使用计算机模拟ADMET工具评估了BCS2的类药性质。
BCS2对MCF - 7细胞表现出显著的细胞毒性活性，其次是MDA - MB - 231和MDA - MB - 468细胞，其半数抑制浓度（IC50）分别为2.368μM、4.843μM和6.472μM，且不影响正常乳腺细胞MCF - 10A。在DMBA诱导的动物模型中，BCS2表现出强大的抗肿瘤潜力，并对荷瘤动物体内的内源性酶促和非酶促抗氧化剂具有保护作用。通过组织病理学和场发射扫描电子显微镜（FESEM）分析，观察到从实验动物切除的乳腺/肿瘤组织的细胞结构和超微结构有明显改善。BCS2处理后，观察到抗氧化蛋白Keap1和HO - 1显著上调，炎症蛋白MAPK和NF - κB下调。计算机模拟计算研究预测了BCS2与Keap1、HO - 1和MAPK蛋白活性口袋的强效结合，这验证了生物学研究结果。
该研究揭示了BCS2通过Nrf2 - Keap1 - HO - 1信号通路对乳腺癌具有强大的抗氧化和抗肿瘤潜力。
