新型表皮生长因子受体抑制剂的计算评估与体外验证。
Computational Evaluation and In Vitro Validation of New Epidermal Growth Factor Receptor Inhibitors.
机构信息
ProtoQSAR SL, European Center for Innovative Companies (CEEI), Valencia Technology Park, Avenida Benjamin Franklin 12, 46980 Paterna, Valencia, Spain.
Department of Medical Oncology, Institute of Biomedical Research INCLIVA, University of Valencia, Valencia, Spain.
出版信息
Curr Top Med Chem. 2020;20(18):1628-1639. doi: 10.2174/1568026620666200603122726.
BACKGROUND
The Epidermal Growth Factor Receptor (EGFR) is a transmembrane protein that acts as a receptor of extracellular protein ligands of the epidermal growth factor (EGF/ErbB) family. It has been shown that EGFR is overexpressed by many tumours and correlates with poor prognosis. Therefore, EGFR can be considered as a very interesting therapeutic target for the treatment of a large variety of cancers such as lung, ovarian, endometrial, gastric, bladder and breast cancers, cervical adenocarcinoma, malignant melanoma and glioblastoma.
METHODS
We have followed a structure-based virtual screening (SBVS) procedure with a library composed of several commercial collections of chemicals (615,462 compounds in total) and the 3D structure of EGFR obtained from the Protein Data Bank (PDB code: 1M17). The docking results from this campaign were then ranked according to the theoretical binding affinity of these molecules to EGFR, and compared with the binding affinity of erlotinib, a well-known EGFR inhibitor. A total of 23 top-rated commercial compounds displaying potential binding affinities similar or even better than erlotinib were selected for experimental evaluation. In vitro assays in different cell lines were performed. A preliminary test was carried out with a simple and standard quick cell proliferation assay kit, and six compounds showed significant activity when compared to positive control. Then, viability and cell proliferation of these compounds were further tested using a protocol based on propidium iodide (PI) and flow cytometry in HCT116, Caco-2 and H358 cell lines.
RESULTS
The whole six compounds displayed good effects when compared with erlotinib at 30 μM. When reducing the concentration to 10μM, the activity of the 6 compounds depends on the cell line used: the six compounds showed inhibitory activity with HCT116, two compounds showed inhibition with Caco-2, and three compounds showed inhibitory effects with H358. At 2 μM, one compound showed inhibiting effects close to those from erlotinib.
CONCLUSION
Therefore, these compounds could be considered as potential primary hits, acting as promising starting points to expand the therapeutic options against a wide range of cancers.
背景
表皮生长因子受体（EGFR）是一种跨膜蛋白，作为表皮生长因子（EGF/ErbB）家族的细胞外蛋白配体的受体。已经表明，许多肿瘤过度表达 EGFR，并且与预后不良相关。因此，EGFR 可以被认为是治疗多种癌症的非常有前途的治疗靶点，如肺癌、卵巢癌、子宫内膜癌、胃癌、膀胱癌和乳腺癌、宫颈腺癌、恶性黑色素瘤和胶质母细胞瘤。
方法
我们采用基于结构的虚拟筛选（SBVS）程序，该程序使用了由多个商业化学库组成的库（总共 615462 种化合物）和从蛋白质数据银行（PDB 代码：1M17）获得的 EGFR 的 3D 结构。根据这些分子与 EGFR 的理论结合亲和力对该运动的对接结果进行排序，并与 EGFR 的已知抑制剂厄洛替尼的结合亲和力进行比较。选择了总共 23 种评分最高的商业化合物，这些化合物显示出与厄洛替尼相似甚至更好的潜在结合亲和力，用于实验评估。在不同的细胞系中进行了体外测定。使用简单和标准的快速细胞增殖测定试剂盒进行了初步测试，其中六种化合物与阳性对照相比表现出显著的活性。然后，使用基于碘化丙啶（PI）和在 HCT116、Caco-2 和 H358 细胞系中使用流式细胞术的方案进一步测试了这些化合物的细胞活力和增殖。
结果
与 30μM 时的厄洛替尼相比，这六种化合物整体均显示出良好的效果。当浓度降低至 10μM 时，六种化合物的活性取决于所用的细胞系：六种化合物对 HCT116 具有抑制活性，两种化合物对 Caco-2 具有抑制活性，三种化合物对 H358 具有抑制活性。在 2μM 时，一种化合物表现出接近厄洛替尼的抑制作用。
结论
因此，这些化合物可被视为潜在的主要命中物，可作为针对广泛癌症的治疗选择的有前途的起点。
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