In Vitro and In Silico Evaluation of Bikaverin as a Potent Inhibitor of Human Protein Kinase CK2
In Vitro and In Silico Evaluation of Bikaverin as a Potent Inhibitor of Human Protein Kinase CK2
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DOI：
10.3390/molecules24071380
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发表时间：
2019-04-08
期刊：
影响因子：
4.6
通讯作者：
Jose, Joachim
中科院分区：
文献类型：
--
作者：
Haidar, Samer;Aichele, Dagmar;Jose, Joachim
蛋白激酶 CK2 是人类疾病（特别是癌症）治疗干预的新兴靶点。该酶的抑制剂目前正在进行临床试验，表明人类 CK2 的成药性。通过对ZINC数据库的虚拟筛选，我们发现天然化合物比卡维林能够很好地契合于靶酶CK2的ATP结合位点。通过使用 CK2 全酶进行进一步的体外评估，比卡维林成为一种有效的抑制剂，IC50 值为 1.24 μM。在这项工作中，使用 Caco-2 细胞渗透性测定法测定比卡维林的细胞渗透性，作为细胞评估的先决条件，结果表明该化合物具有细胞渗透性，P-app 值为 4.46 x 10(-6) cm/s。使用 MTT 测定法测试比卡维林对细胞活力的影响，并使用 EdU 测定法在不同的癌细胞系（MCF7、A427 和 A431 细胞）中测试其对细胞增殖的影响。用10μM比卡维林处理24小时后，细胞活力和细胞增殖显着降低。此外，IncuCyteA (R) 活细胞成像系统用于监测比卡维林在三种测试的癌细胞系中的细胞毒性。最后，进行分子动力学研究以阐明比卡维林在 CK2 ATP 结合位点的配体结合模式并鉴定所涉及的氨基酸。
Protein kinase CK2 is an emerging target for therapeutic intervention in human diseases, particularly in cancer. Inhibitors of this enzyme are currently in clinical trials, indicating the druggability of human CK2. By virtual screening of the ZINC database, we found that the natural compound bikaverin can fit well in the ATP binding site of the target enzyme CK2. By further in vitro evaluation using CK2 holoenzyme, bikaverin turned to be a potent inhibitor with an IC50 value of 1.24 mu M. In this work, the cell permeability of bikaverin was determined using a Caco-2 cell permeability assay as a prerequisite for cellular evaluation and the compound turned out to be cell permeable with a P-app- value of 4.46 x 10(-6) cm/s. Bikaverin was tested for its effect on cell viability using a MTT assay and cell proliferation using an EdU assay in different cancer cell lines (MCF7, A427 and A431 cells). Cell viability and cell proliferation were reduced dramatically after treatment with 10 mu M bikaverin for 24 h. Additionally the IncuCyteA (R) live-cell imaging system was applied for monitoring the cytotoxicity of bikaverin in the three tested cancer cell lines. Finally, molecular dynamic studies were performed to clarify the ligand binding mode of bikaverin at the ATP binding site of CK2 and to identify the amino acids involved.