Phenylpyrazolo[3,4-d]pyrimidine is considered a milestone scaffold known to possess various biological activities such as antiparasitic, antifungal, antimicrobial, and antiproliferative activities. In addition, the urgent need for selective and potent novel anticancer agents represents a major route in the drug discovery process. Herein, new aryl analogs were synthesized and evaluated for their anticancer effects on a panel of cancer cell lines: MCF-7, HCT116, and HePG-2. Some of these compounds showed potent cytotoxicity, with variable degrees of potency and cell line selectivity in antiproliferative assays with low resistance. As the analogs carry the pyrazolopyrimidine scaffold, which looks structurally very similar to tyrosine and receptor kinase inhibitors, the potent compounds were evaluated for their inhibitory effects on three essential cancer targets: EGFRWT, EGFRT790M, VGFR2, and Top-II. The data obtained revealed that most of these compounds were potent, with variable degrees of target selectivity and dual EGFR/VGFR2 inhibitors at the IC50 value range, i.e., 0.3–24 µM. Among these, compound 5i was the most potent non-selective dual EGFR/VGFR2 inhibitor, with inhibitory concentrations of 0.3 and 7.60 µM, respectively. When 5i was tested in an MCF-7 model, it effectively inhibited tumor growth, strongly induced cancer cell apoptosis, inhibited cell migration, and suppressed cell cycle progression leading to DNA fragmentation. Molecular docking studies were performed to explore the binding mode and mechanism of such compounds on protein targets and mapped with reference ligands. The results of our studies indicate that the newly discovered phenylpyrazolo[3,4-d]pyrimidine-based multitarget inhibitors have significant potential for anticancer treatment.

中文翻译：

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对接研究支持开发具有有效抗癌活性和多靶点酶抑制作用的新型苯基吡唑并[3,4-d]嘧啶类似物

苯基吡唑并[3,4-d]嘧啶被认为是一种里程碑式的支架，已知具有多种生物活性，例如抗寄生虫、抗真菌、抗菌和抗增殖活性。此外，对选择性和有效的新型抗癌药物的迫切需求是药物发现过程中的一个主要途径。在此，合成了新的芳基类似物，并评估了它们对一组癌细胞系：MCF-7、HCT116 和 HePG-2 的抗癌作用。其中一些化合物表现出强大的细胞毒性，在低耐药性的抗增殖测定中具有不同程度的效力和细胞系选择性。由于这些类似物带有吡唑并嘧啶支架，其结构看起来与酪氨酸和受体激酶抑制剂非常相似，因此评估了这些有效化合物对三个重要癌症靶点的抑制作用：EGFRWT、EGFRT790M、VGFR2 和 Top-II。获得的数据显示，大多数这些化合物都是有效的，具有不同程度的靶点选择性，并且是 IC50 值范围（即 0.3–24 µM）的双重 EGFR/VGFR2 抑制剂。其中，化合物5i是最有效的非选择性EGFR/VGFR2双重抑制剂，抑制浓度分别为0.3和7.60 µM。当 5i 在 MCF-7 模型中进行测试时，它有效抑制肿瘤生长，强烈诱导癌细胞凋亡，抑制细胞迁移，并抑制导致 DNA 断裂的细胞周期进程。进行分子对接研究以探索此类化合物与蛋白质靶标的结合模式和机制，并与参考配体进行定位。我们的研究结果表明，新发现的基于苯基吡唑并[3,4-d]嘧啶的多靶点抑制剂具有显着的抗癌治疗潜力。