Khanam Rashmin, Ahmad Kamal, Hejazi Iram I, Siddique Ibrar A, Kumar Vikash, Bhat Abdul Roouf, Azam Amir, Athar Fareeda
Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi, 110025, India.
National Institute of Immunology, New Delhi, 110067, India.
Cancer Chemother Pharmacol. 2017 Nov;80(5):1027-1042. doi: 10.1007/s00280-017-3414-6. Epub 2017 Aug 16.
Cancer has become one of the global health issues and it is the life-threatening disease characterized by unrestrained growth of cells. Despite various advances being adopted by chemotherapeutic management, the use of the current anticancer drugs such as Doxorubicin, Asparginase, Methotrexate, Vincristine remains limited due to high toxicity, side effects and developing drug resistance. Apoptosis is a crucial cellular process and improper regulation of apoptotic signaling pathways may lead to cancer formation. Subsequently, the synthesis of effective chemotherapeutic agents that can induce apoptosis in tumor cell has emerged as a significant approach in cancer drug discovery.
The goal of this work is to develop a potential antitumor agent exerting significant inhibitory effects on cancer cell and low cytotoxicity, for which we focused on the structural features of 1,3,4-oxadiazoles as it a privileged scaffold in modern medicinal chemistry and have the ability to inhibit growth factors, enzymes and kinases potentially involved in the attainment of cellular immortality and carcinogenesis.
In vitro MTT screening assay showed the compound 5-aminophenyl-2-butylthio-1,3,4-oxadiazole (5e) showing the highest inhibitory effect against MCF-7 cancer cell with IC value 10.05 ± 1.08 µM while it is much safer and less toxic on normal cell line (HEK-293). The dose-dependent treatment of MCF-7 cells with 5e resulted in inhibition of cell migration in the wound healing assay. The flow-cytometry analysis showed the cells arrested in G0/G1 phase of the cell cycle. Compound 5e induced apoptosis of MCF-7 cells was characterized using DAPI staining and Annexin V-PE/7-AAD dual binding assay. Reduction of NBT by compound 5e showed a reduced generation of ROS. Western blotting studies showed high activation of apoptotic protein Caspase3 and decrease in expression of anti-apoptotic protein BCL-2.
Based on the results of in vitro studies, it could be concluded that compound 5e showed a significant inhibitory growth effect on MCF-7 cells and have the potential to be developed as lead molecule and further structural modifications may result in promising new anticancer agents.
癌症已成为全球健康问题之一，是一种以细胞无节制生长为特征的危及生命的疾病。尽管化疗管理取得了各种进展，但由于毒性高、副作用大以及产生耐药性，目前使用的抗癌药物如多柔比星、天冬酰胺酶、甲氨蝶呤、长春新碱的应用仍然有限。细胞凋亡是一个关键的细胞过程，凋亡信号通路的不当调节可能导致癌症形成。随后，能够诱导肿瘤细胞凋亡的有效化疗药物的合成已成为癌症药物发现中的一种重要方法。
这项工作的目标是开发一种对癌细胞具有显著抑制作用且细胞毒性低的潜在抗肿瘤药物，为此我们关注了1,3,4-恶二唑的结构特征，因为它是现代药物化学中的一个优势骨架，并且有能力抑制可能参与细胞永生化和致癌过程的生长因子、酶和激酶。
体外MTT筛选试验表明，化合物5-氨基苯基-2-丁硫基-1,3,4-恶二唑（5e）对MCF-7癌细胞的抑制作用最高，IC值为10.05±1.08μM，而对正常细胞系（HEK-293）则更安全、毒性更小。用5e对MCF-7细胞进行剂量依赖性处理导致伤口愈合试验中细胞迁移受到抑制。流式细胞术分析表明细胞停滞在细胞周期的G0/G1期。使用DAPI染色和膜联蛋白V-PE/7-AAD双重结合试验对化合物5e诱导MCF-7细胞凋亡进行了表征。化合物5e使NBT还原表明活性氧的产生减少。蛋白质印迹研究表明凋亡蛋白Caspase3高度活化，抗凋亡蛋白BCL-2的表达降低。
基于体外研究结果，可以得出结论，化合物5e对MCF-7细胞显示出显著的抑制生长作用，并有可能被开发为先导分子，进一步的结构修饰可能会产生有前景的新型抗癌药物。
