Design and discovery of new antiproliferative 1,2,4-triazin-3(2H)-ones as tubulin polymerization inhibitors targeting colchicine binding site
Design and discovery of new antiproliferative 1,2,4-triazin-3(2H)-ones as tubulin polymerization inhibitors targeting colchicine binding site
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DOI：
10.1016/j.bioorg.2021.104965
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发表时间：
2021-05-18
影响因子：
5.1
通讯作者：
Beauchemin, Andre M.
中科院分区：
文献类型：
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作者：
Eissa, Ibrahim H.;Dahab, Mohammed A.;Beauchemin, Andre M.
以1，2，4-三嗪-3（2 H）-酮为基础，设计并合成了35个新的秋水仙碱结合位点抑制剂。这些分子是通过容易获得的α-氨基酮和作为掩蔽的N-异氰酸酯前体的肼基甲酸苯酯之间的级联反应合成的。合成的衍生物是顺式限制的考布他汀A4类似物，含有1，2，4-三嗪-3（2 H）-酮代替烯键，并且它们具有秋水仙碱结合位点抑制剂的基本药效学特征。使用秋水仙碱作为阳性对照，在体外评价合成的化合物对一组三种人癌细胞系（MCF-7、HepG-2和HCT-116）的抗增殖活性。其中，两个化合物5i和6 i表现出显着的抗增殖作用对所有细胞系的IC 50范围从8.2 - 18.2 μ M。对作为微管蛋白聚合抑制剂的最具活性的细胞毒性剂进行了进一步的研究。化合物5i和6 i有效地抑制微管组装，IC 50值为3.9 - 7.8 μ M。发现微管蛋白聚合测定结果与细胞毒性结果相当。细胞周期分析显示类似物5i在HepG-2细胞中显著的G2/M细胞周期阻滞。最具活性的化合物4 i、4j、5g、5i和6 i在正常人肺细胞W1 -38中不诱导显著的细胞死亡，表明它们对癌细胞的选择性。此外，与对照相比，这些化合物上调了活性半胱天冬酶-3的水平，并使促凋亡蛋白Bax的水平提高了5至7倍。此外，对化合物5i进行细胞凋亡分析以评估其细胞凋亡潜力。最后，进行计算机模拟研究以揭示与秋水仙碱结合位点的可能相互作用。对所设计的化合物进行了ADME预测研究，结果表明，所设计的化合物不仅具有良好的微管蛋白聚合抑制活性，而且具有良好的药代动力学和药物相似性。
Thirty-five new colchicine binding site inhibitors have been designed and synthesized based on the 1,2,4-triazin3(2H)-one nucleus. Such molecules were synthesized through a cascade reaction between readily accessible alpha-amino ketones and phenyl carbazate as a masked N-isocyanate precursor. The synthesized derivatives are cisoid restricted combretastatin A4 analogues containing 1,2,4-triazin-3(2H)-one in place of the olefinic bond, and they have the same essential pharmacophoric features of colchicine binding site inhibitors. The synthesized compounds were evaluated in vitro for their antiproliferative activities against a panel of three human cancer cell lines (MCF-7, HepG-2, and HCT-116), using colchicine as a positive control. Among them, two compounds 5i and 6i demonstrated a significant antiproliferative effect against all cell lines with IC50 ranging from 8.2 18.2 mu M. Further investigation was carried out for the most active cytotoxic agents as tubulin polymerization inhibitors. Compounds 5i and 6i effectively inhibited microtubule assembly with IC50 values ranging from 3.9 to 7.8 mu M. Tubulin polymerization assay results were found to be comparable with the cytotoxicity results. The cell cycle analysis revealed significant G2/M cell cycle arrest of the analogue 5i in HepG-2 cells. The most active compounds 4i, 4j, 5 g, 5i and 6i did not induce significant cell death in normal human lung cells Wl-38, suggesting their selectivity against cancer cells. Also, These compounds upregulated the level of active caspase-3 and boosted the levels of the pro-apoptotic protein Bax by five to seven folds in comparison to the control. Moreover, apoptosis analyses were conducted for compound 5i to evaluate its apoptotic potential. Finally, in silico studies were conducted to reveal the probable interaction with the colchicine binding site. ADME prediction study of the designed compounds showed that they are not only with promising tubulin polymerization inhibitory activity but also with favorable pharmacokinetic and drug-likeness properties.