Department of Pharmaceutical Chemistry, Institute of Pharmacy, Nirma University, Ahmedabad 382 481, India.
Department of Life Sciences, Gujarat University, Ahmedabad 380 009, India.
Bioorg Chem. 2021 Jan;106:104501. doi: 10.1016/j.bioorg.2020.104501. Epub 2020 Nov 24.
Lung cancer is one of the most prevailed cancer worldwide. Many genes get mutated in lung cancer but the involvement of EGFR, KRAS, PTEN and PIK3CA are more common. Unavailability of potent drugs and resistance to the available drugs are major concern in the treatment of lung cancer. In the present research, mTOR was selected as an important alternative target for the treatment of lung cancer which involves the PI3K/AKT/mTOR pathway. We studied binding interactions of AZD-2014 with the mTOR protein to identify important interactions required to design potent mTOR inhibitors which was supported by QSAR studies. Pharmacophore based virtual screening studies provided core scaffold, THQ. Based on molecular docking interactions, 31 THQ derivatives were synthesized and characterized. All compounds were screened for cellular mTOR enzyme assay along with antiproliferative activity against the panel of cancerous cell lines, from which 6 compounds were further screened for colony forming assay. Two most potent compounds, HB-UC-1 and HB-UC-5, were further screened for flow cytometry analysis, gene expression study and western blot analysis. Gene expression study revealed the efficiency of compound HB-UC-1 against both mTORC1 and mTORC2 by affecting downstream regulators of mTORC1 (EBP, eIFEBP) and mTORC2 (PCK1), respectively. In western blot analysis, both compounds, inhibited phosphorylation of AKT S473 which proved the efficiency these compounds against the mTORC2. These two compounds were further screened for in-vivo biological evaluation. Both compounds increased lifespan of cancer-bearing animals with improvement in mean survival time. Further, in bezopyrene induced lung cancer animal model, both compounds showed effectiveness through the biochemical parameters and histopathological evaluation of the lung tissue. In future, potent hit compound from this series could be modified to develop lead mTOR inhibitors for the treatment of lung cancer.
肺癌是全球最常见的癌症之一。许多基因在肺癌中发生突变，但 EGFR、KRAS、PTEN 和 PIK3CA 的参与更为常见。缺乏有效的药物和对现有药物的耐药性是肺癌治疗的主要关注点。在目前的研究中，mTOR 被选为治疗肺癌的重要替代靶标，涉及 PI3K/AKT/mTOR 途径。我们研究了 AZD-2014 与 mTOR 蛋白的结合相互作用，以确定设计有效的 mTOR 抑制剂所需的重要相互作用，这得到了 QSAR 研究的支持。基于药效基团的虚拟筛选研究提供了核心支架 THQ。基于分子对接相互作用，合成并表征了 31 个 THQ 衍生物。所有化合物均进行了细胞 mTOR 酶测定筛选，并对一组癌细胞系进行了抗增殖活性筛选，从中筛选出 6 种化合物进行集落形成测定。两种最有效的化合物 HB-UC-1 和 HB-UC-5 进一步进行了流式细胞术分析、基因表达研究和 Western blot 分析。基因表达研究表明，化合物 HB-UC-1 通过影响 mTORC1 的下游调节剂（EBP、eIFEBP）和 mTORC2 的下游调节剂（PCK1），对 mTORC1 和 mTORC2 均有效。在 Western blot 分析中，两种化合物均抑制了 AKT S473 的磷酸化，证明了这些化合物对 mTORC2 的有效性。这两种化合物进一步进行了体内生物学评价。这两种化合物都增加了荷瘤动物的寿命，提高了平均生存时间。此外，在苯并芘诱导的肺癌动物模型中，两种化合物均通过肺组织的生化参数和组织病理学评估显示出有效性。在未来，该系列中的有效化合物可以进行修饰，以开发用于治疗肺癌的有效的 mTOR 抑制剂。
