Medical Laboratory Department, College of Applied Medical Sciences, Taibah University, Yanbu, Saudi Arabia.
Chemistry Department, Faculty of Science, Suez Canal University, Ismailia, Egypt.
J Steroid Biochem Mol Biol. 2020 Apr;198:105604. doi: 10.1016/j.jsbmb.2020.105604. Epub 2020 Jan 23.
Worldwide, cancer is still an area with high unmet medical need. Lead optimization efforts towards structure-based drug design were employed to discover newly synthesized hetero-steroid derivatives with promising anticancer effects against hepatocellular carcinoma (HCC). The aim of our study is to evaluate the anti-proliferative activity and the mechanism, a dual PI3K/mTOR inhibitor, and mechanism of action of a series of heterocylic androstane derivatives as anti-HCC agent. The cytotoxic effects of different heterocylic androstanes and 5FU as single agents, were assessed against both HepG2 cells and Non-malignant MDCK cell line to assess the toxicity. Then the underlying mechanism of compound 4 as most promising compound was evaluated using molecular docking, MTT assay, cell cycle analysis, DNA fragmentation, and real-time PCR. The results of MTT assay showed potential cytotoxic effect for compound 4 and 5 against liver cancer cell line with IC value 39.81 and 57.54 μM, respectively. Inhibition of the PI3K/AKT/mTOR pathway was achieved by compound 4, which was documented by molecular docking and augmented by gene expression analysis. Detailed mechanism revealed that compound 4 induced cell cycle arrest, DNA fragmentation, and induction of apoptosis by inhibition of anti-apoptotic genes, and upregulation of apoptotic genes. Our results shed a light on aminopyrazoloandrostane derivative 4 as an inhibitor of the PI3K/AKT/mTOR pathway, which might be acting as promising anti-liver cancer agent. Our data support further investigation of agents targeting the PI3K/AKT/mTOR.
在全球范围内，癌症仍然是一个医学需求未得到满足的领域。我们采用基于结构的药物设计方法进行先导化合物优化，旨在发现具有潜在抗肝癌（HCC）作用的新型杂甾体衍生物。本研究旨在评估一系列杂环雄甾烷衍生物作为抗 HCC 药物的抗增殖活性和作用机制，即双重 PI3K/mTOR 抑制剂及其作用机制。我们评估了不同杂环雄甾烷和 5FU 作为单一药物对 HepG2 细胞和非恶性 MDCK 细胞系的细胞毒性作用，以评估其毒性。然后，我们使用分子对接、MTT 测定、细胞周期分析、DNA 片段化和实时 PCR 评估了最有前途的化合物 4 的潜在作用机制。MTT 测定结果表明，化合物 4 和 5 对肝癌细胞系具有潜在的细胞毒性作用，IC 值分别为 39.81 和 57.54 μM。化合物 4 通过抑制 PI3K/AKT/mTOR 通路，实现了对该通路的抑制，这一结果通过分子对接得到了证实，并通过基因表达分析得到了增强。详细的机制表明，化合物 4 通过抑制抗凋亡基因和上调凋亡基因，诱导细胞周期停滞、DNA 片段化和凋亡。我们的研究结果表明，氨基吡唑并雄烷衍生物 4 可能是一种抑制 PI3K/AKT/mTOR 通路的抑制剂，可作为有前途的抗肝癌药物。我们的数据支持进一步研究针对 PI3K/AKT/mTOR 的药物。
