The strategic planning of this study is based upon using the nanoformulation method to prepare nanoparticles 4-SLNs and 4-LPHNPs of the previously prepared 4,5-diphenyl-1H-pyrazolo[3,4-c]pyridazin-3-amine (4) after confirming its structure with single crystal X-ray analysis. These nanoparticles exhibited promising cytotoxic activity against HepG-2, HCT-116 and MCF-7 cancer cell lines in comparison with the reference doxorubicin and the original derivative 4. Moreover, their inhibitory assessment against EGFR and CDK-2/cyclin A2 displayed improved and more favorable impact than the parent 4 and the references. Detection of their influence upon cancer biomarkers revealed upregulation of Bax, p53 and caspase-3 levels and downregulation of Bcl-2 levels. The docking simulation demonstrated that the presence of the pyrazolo[3,4-c]pyridazin-3-amine scaffold is amenable to enclosure and binding well within EGFR and CDK-2 receptors through different hydrophilic interactions. The pharmacokinetic and physicochemical properties of target 4 were also assessed with ADME investigation, and the outcome indicated good drug-like characteristics.

中文翻译：

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作为潜在 EGFR 和 CDK-2 抑制剂的吡唑并哒嗪衍生物纳米颗粒：设计、结构测定、抗癌评估和计算机研究。

本研究的战略规划是基于使用纳米制剂方法制备先前制备的4,5-二苯基-1H-吡唑并[3,4-c]哒嗪-3-胺（4 ）通过单晶X射线分析确认其结构后。与参考阿霉素和原始衍生物 4 相比，这些纳米颗粒对 HepG-2、HCT-116 和 MCF-7 癌细胞系表现出有希望的细胞毒活性。此外，它们对 EGFR 和 CDK-2/cyclin A2 的抑制评估显示出改善和改善。比母体4和参考文献产生更有利的影响。检测它们对癌症生物标志物的影响揭示了 Bax、p53 和 caspase-3 水平的上调以及 Bcl-2 水平的下调。对接模拟表明，吡唑并[3,4-c]哒嗪-3-胺支架的存在易于通过不同的亲水相互作用在EGFR和CDK-2受体内封装和结合。还通过 ADME 研究评估了靶标 4 的药代动力学和理化性质，结果表明具有良好的药物样特性。