Department of Biological Sciences, Sanghuh College of Life Sciences, Konkuk University, Seoul, 05029, Republic of Korea.
Cancer and Metabolism Institute, Konkuk University, Seoul, 05029, Republic of Korea.
Daru. 2019 Jun;27(1):265-281. doi: 10.1007/s40199-019-00272-5. Epub 2019 Jun 1.
Several 4,6-diarylpyrimidin-2-amine derivatives show anticancer properties. However, their mode of action is not fully characterized. To develop potent anticancer chemotherapeutic agents, we designed and synthesized 25 4,6-diphenylpyrimidin-2-amine derivatives containing a guanidine moiety.
Clonogenic long-term survival assays were performed to screen anticancer compounds. To derive the structural conditions showing good cytotoxicities against cancer cells, quantitative structure-activity relationships (QSAR) were calculated. Biological activities were determined by flow cytometry for cell cycle analysis and by immunoblot analysis for the detection of Aurora kinase A (AURKA) activity. Because 2-(2-Amino-6-(2,4-dimethoxyphenyl)pyrimidin-4-yl) phenol (derivative 12) selectively inhibited AURKA activity from the kinome assay, in silico docking experiments were performed to elucidate the molecular binding mode between derivative 12 and AURKA.
The pharmacophores were derived based on the QSAR calculations. Derivative 12 inhibited AURKA activity and reduced phosphorylation of AURKA at Thr283 in HCT116 human colon cancer cells. Derivative 12 caused the accumulation of the G2/M phase of the cell cycle and triggered the cleavages of caspase-3, caspase -7, and poly(ADP-ribose) polymerase. The binding energies of 30 apo-AURKA - derivative 12 complexes obtained from in silico docking ranged from -16.72 to -11.63 kcal/mol.
Derivative 12 is an AURKA inhibitor, which reduces clonogenicity, arrests the cell cycle at the G2/M phase, and induces caspase-mediated apoptotic cell death in HCT116 human colon cancer cells. In silico docking demonstrated that derivative 12 binds to AURKA well. The structure-activity relationship calculations showed hydrophobic substituents and 1-naphthalenyl group at the R2 position increased the activity. The existence of an H-bond acceptor at C-2 of the R1 position increased the activity, too. Graphical abstract Derivative 12 inhibits Aurora kinase A activity and causes the G2/M phase arrest of the cell cycle.
几种 4,6-二芳基嘧啶-2-胺衍生物具有抗癌特性。然而，它们的作用模式尚未完全阐明。为了开发有效的抗癌化学治疗药物，我们设计并合成了 25 种含有胍基部分的 4,6-二苯基嘧啶-2-胺衍生物。
采用集落形成长期存活实验筛选抗癌化合物。为了推导出对癌细胞具有良好细胞毒性的结构条件，进行了定量构效关系（QSAR）计算。通过流式细胞术进行细胞周期分析和免疫印迹分析检测 Aurora 激酶 A（AURKA）活性来确定生物活性。由于 2-(2-氨基-6-(2,4-二甲氧基苯基)嘧啶-4-基)苯酚（衍生物 12）在激酶组实验中选择性抑制 AURKA 活性，因此进行了计算机对接实验以阐明衍生物 12 与 AURKA 之间的分子结合模式。
根据 QSAR 计算推导出药效团。衍生物 12 抑制 AURKA 活性并降低 HCT116 人结肠癌细胞中 AURKA 的 Thr283 磷酸化。衍生物 12 导致细胞周期 G2/M 期的积累，并触发 caspase-3、caspase-7 和聚（ADP-核糖）聚合酶的裂解。从计算机对接获得的 30 个 apo-AURKA-衍生物 12 复合物的结合能范围为-16.72 至-11.63 kcal/mol。
衍生物 12 是一种 AURKA 抑制剂，可降低集落形成能力，使细胞周期在 G2/M 期停滞，并诱导 HCT116 人结肠癌细胞中 caspase 介导的凋亡细胞死亡。计算机对接表明，衍生物 12 与 AURKA 结合良好。构效关系计算表明，R2 位的疏水性取代基和 1-萘基增加了活性。R1 位 C-2 上存在氢键受体也增加了活性。
