Novel anti-proliferative agents possessing pyrimidine scaffolds were designed, synthesized and evaluated for their IC50 values using MTT assay. Most compounds displayed good to excellent activity against the two tested breast cancer lines (MCF-7 and MDA-MB-231) as compared to 5-FU. The observed IC50 values for active compounds ranged from 0.27 to 10.57 µM in MCF-7 compared to the reference drug 5-FU (IC50 = 10.80 µM) and from 0.73 to 29.07 µM in MDA-MB-231 (IC50 for 5-FU = 11.40 µM). SAR analysis indicated that compounds 2c, 3b with hydrazone functionalities and compound 12 possessing pyrazolone ring exhibited superior activities. The most promising compounds were evaluated for their inhibitory activity against epidermal growth factor receptor (EGFR) and aromatase (ARO) enzymes and were further tested for caspase-9 activation, apoptosis and Annexin V/PI staining. Results of enzyme-based experiments indicated that the tested compounds 2c and 12 exert their activities through EGFR inhibition while compound 3b exhibited remarkable ARO inhibition activity. Furthermore, they remarkably induce caspase-9 activation and showed pre G1 apoptosis and cell cycle arrest at G2/M phase. In addition, docked compounds displayed good binding affinities to the target enzymes. Binding interaction details for the most promising inhibitors with the active site of the target enzymes EGFR and ARO utilizing MOE-dock method are also reported.

中文翻译：

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2-苯胺基嘧啶衍生物：设计，合成，体外抗增殖活性，EGFR和ARO抑制活性，细胞周期分析和分子对接研究。

设计，合成具有嘧啶支架的新型抗增殖剂，并使用MTT分析评估其IC50值。与5-FU相比，大多数化合物对两种测试的乳腺癌细胞系（MCF-7和MDA-MB-231）表现出良好或优异的活性。与参比药物5-FU（IC50 = 10.80 µM）相比，在MCF-7中观察到的活性化合物的IC50值范围为0.27至10.57 µM，在MDA-MB-231中对于5-FU = IC50值则为0.73至29.07 µM。 11.40 µM）。SAR分析表明，具有官能团的化合物2c，3b和具有吡唑啉酮环的化合物12均表现出优异的活性。评估了最有前途的化合物对表皮生长因子受体（EGFR）和芳香化酶（ARO）酶的抑制活性，并进一步测试了caspase-9的激活，细胞凋亡和膜联蛋白V / PI染色。基于酶的实验结果表明，受试化合物2c和12通过EGFR抑制发挥其活性，而化合物3b则表现出显着的ARO抑制活性。此外，它们显着诱导caspase-9激活并显示前G1凋亡和细胞周期停滞在G2 / M期。另外，对接的化合物对靶酶表现出良好的结合亲和力。还报道了最有希望的抑制剂与利用MOE-dock方法的目标酶EGFR和ARO的活性位点的结合相互作用的详细信息。它们显着诱导caspase-9活化，并显示前G1凋亡和细胞周期停滞在G2 / M期。另外，对接的化合物对靶酶表现出良好的结合亲和力。还报道了最有希望的抑制剂与利用MOE-dock方法的目标酶EGFR和ARO的活性位点的结合相互作用的详细信息。它们显着诱导caspase-9活化，并显示前G1凋亡和细胞周期停滞在G2 / M期。另外，对接的化合物对靶酶表现出良好的结合亲和力。还报道了最有希望的抑制剂与利用MOE-dock方法的目标酶EGFR和ARO的活性位点的结合相互作用的详细信息。