Department of Pharmaceutical Organic Chemistry, Faculty of Pharmacy, Mansoura University, Mansoura P.O. Box 35516, Egypt.
Department of Biochemistry and Molecular Genetics, University of Colorado Denver, Denver, CO 80204, USA.
Molecules. 2022 Aug 17;27(16):5245. doi: 10.3390/molecules27165245.
Benzimidazole derivatives are known to be key players in the development of novel anticancer agents. Herein, we aimed to synthesize novel derivatives to target breast cancer. A new series of benzimidazole derivatives conjugated with either six- and five-membered heterocyclic ring or pyrazanobenzimidazoles and pyridobenzimidazole linkers were synthesized yielding compounds - and -, respectively. Structure elucidation of the newly synthesized compounds was achieved through microanalytical analyses and different spectroscopic techniques (H, C-APT and H-H COSY and IR) in addition to mass spectrometry. A biological study for the newly synthesized compounds was performed against breast cancer cell lines (MCF-7), and the most active compounds were further subjected to normal Human lung fibroblast (WI38) which indicates their safety. It was found that most of them exhibit high cytotoxic activity against breast cancer (MCF-7) and low cytotoxic activity against normal (WI38) cell lines. Compounds , , and , which possess the highest anti-breast cancer activity against the MCF-7 cell line, were selected for Pin1 inhibition assay using tannic acid as a reference drug control. Compound was examined for its effect on cell cycle progression and its ability to apoptosis induction. Mechanistic evaluation of apoptosis induction was demonstrated by triggering intrinsic apoptotic pathways via inducing ROS accumulation, increasing Bax, decreasing Bcl-2, and activation of caspases 6, 7, and 9. Binding to 15N-labeled Pin1 enzyme was performed using state-of-the-art N-H HSQC NMR experiments to describe targeting breast cancer on a molecular level. In conclusion, the NMR results demonstrated chemical shift perturbation (peak shifting or peak disappearance) upon adding compound 12 indicating potential binding. Molecular docking using 'Molecular Operating Environment' software was extremely useful to elucidate the binding mode of active derivatives via hydrogen bonding.
苯并咪唑衍生物是新型抗癌药物开发中的关键因素。在此，我们旨在合成新型衍生物以靶向乳腺癌。我们合成了一系列新的苯并咪唑衍生物，它们分别与六元和五元杂环或吡唑并苯并咪唑和吡啶并苯并咪唑连接子共轭。通过微量分析和不同的光谱技术（H、C-APT 和 H-H COSY 以及 IR）以及质谱对新合成化合物的结构进行了阐明。对新合成的化合物进行了针对乳腺癌细胞系（MCF-7）的生物学研究，最活跃的化合物进一步用正常的人肺成纤维细胞（WI38）进行了测试，表明它们是安全的。结果发现，它们中的大多数对乳腺癌（MCF-7）具有很高的细胞毒性活性，而对正常（WI38）细胞系的细胞毒性活性较低。具有最高抗乳腺癌活性的化合物 、 、和 ，被选为 Pin1 抑制测定，使用鞣酸作为参考药物对照。研究了化合物 对细胞周期进程的影响及其诱导细胞凋亡的能力。通过诱导 ROS 积累、增加 Bax、减少 Bcl-2 和激活半胱天冬酶 6、7 和 9 来诱导内在凋亡途径，证明了凋亡诱导的机制评估。使用最先进的 N-H HSQC NMR 实验进行 15N 标记的 Pin1 酶的结合，以描述在分子水平上靶向乳腺癌。总之，NMR 结果表明，添加化合物 12 后会发生化学位移扰动（峰移动或峰消失），表明可能存在结合。使用“分子操作环境”软件进行分子对接非常有助于通过氢键阐明活性衍生物的结合模式。
