Franco Catarina, Kausar Samina, Silva Margarida F B, Guedes Rita C, Falcao Andre O, Brito Maria Alexandra
LASIGE, Department of Informatics, Faculty of Sciences, Universidade de Lisboa, Campo Grande, 1749-016 Lisboa, Portugal.
Research Institute for Medicines, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisboa, Portugal.
Cancers (Basel). 2022 Jul 19;14(14):3506. doi: 10.3390/cancers14143506.
The epidermal growth factor receptor (EGFR) is upregulated in glioblastoma, becoming an attractive therapeutic target. However, activation of compensatory pathways generates inputs to downstream PI3Kp110β signaling, leading to anti-EGFR therapeutic resistance. Moreover, the blood-brain barrier (BBB) limits drugs' brain penetration. We aimed to discover EGFR/PI3Kp110β pathway inhibitors for a multi-targeting approach, with favorable ADMET and BBB-permeant properties. We used quantitative structure-activity relationship models and structure-based virtual screening, and assessed ADMET properties, to identify BBB-permeant drug candidates. Predictions were validated in in vitro models of the human BBB and BBB-glioma co-cultures. The results disclosed 27 molecules (18 EGFR, 6 PI3Kp110β, and 3 dual inhibitors) for biological validation, performed in two glioblastoma cell lines (U87MG and U87MG overexpressing EGFR). Six molecules (two EGFR, two PI3Kp110β, and two dual inhibitors) decreased cell viability by 40-99%, with the greatest effect observed for the dual inhibitors. The glioma cytotoxicity was confirmed by analysis of targets' downregulation and increased apoptosis (15-85%). Safety to BBB endothelial cells was confirmed for three of those molecules (one EGFR and two PI3Kp110β inhibitors). These molecules crossed the endothelial monolayer in the BBB in vitro model and in the BBB-glioblastoma co-culture system. These results revealed novel drug candidates for glioblastoma treatment.
表皮生长因子受体（EGFR）在胶质母细胞瘤中上调，成为一个有吸引力的治疗靶点。然而，补偿途径的激活会产生向下游PI3Kp110β信号传导的输入，导致抗EGFR治疗耐药。此外，血脑屏障（BBB）限制了药物的脑渗透。我们旨在发现用于多靶点方法的EGFR/PI3Kp110β途径抑制剂，具有良好的药物代谢动力学、药效学及毒性性质（ADMET）和血脑屏障穿透特性。我们使用定量构效关系模型和基于结构的虚拟筛选，并评估ADMET性质，以识别具有血脑屏障穿透性的候选药物。预测结果在人血脑屏障体外模型和血脑屏障-胶质瘤共培养模型中得到验证。结果揭示了27种分子（18种EGFR抑制剂、6种PI3Kp110β抑制剂和3种双重抑制剂）用于生物学验证，在两种胶质母细胞瘤细胞系（U87MG和过表达EGFR的U87MG）中进行。六种分子（两种EGFR抑制剂、两种PI3Kp110β抑制剂和两种双重抑制剂）使细胞活力降低了40-99%，其中双重抑制剂的效果最为显著。通过分析靶点下调和凋亡增加（15-85%）证实了对胶质瘤的细胞毒性。其中三种分子（一种EGFR抑制剂和两种PI3Kp110β抑制剂）对血脑屏障内皮细胞的安全性得到证实。这些分子在体外血脑屏障模型和血脑屏障-胶质母细胞瘤共培养系统中穿过内皮单层。这些结果揭示了用于胶质母细胞瘤治疗的新型候选药物。
