Multi-Targeting Approach in Glioblastoma Using Computer-Assisted Drug Discovery Tools to Overcome the Blood–Brain Barrier and Target EGFR/PI3Kp110β Signaling

Abstract

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.

In vitro study of the blood–brain barrier transport of bioactives from Mediterranean foods

“…Despite the complexity of the BBB in vivo , where endothelial cells are entreated by a thick basement membrane and by pericytes and astrocytes end-feet, most BBB in vitro models rely on the use of BMEC, considered to form the anatomic basis of the BBB. 43 Particularly, the cell line here used, HBMEC, has been employed in multiple studies of BBB permeation of natural compounds and of drug candidates, 6,72,73 which attests its scientific recognition and value as a simplified in vitro model of the BBB to predict the potential BBB transport of bioactives.…”

A Drug Screening Reveals Minocycline Hydrochloride as a Therapeutic Option to Prevent Breast Cancer Cells Extravasation across the Blood–Brain Barrier

“…In the present work, a concentration of 0.1 µM decreased the number of cells per field and reduced the size of cell clusters. Moreover, Mol6 and KW-2478, predicted PI3K [ 39 ] and HSP90 inhibitors, respectively, revealed a significant increase in the percentage of cells with aberrant nuclei, at a concentration of 1 µM. Interestingly, the concentrations of KW-2478 effective in other studies (e.g., in reducing tumour cell viability and/or proliferation) were slightly higher (3 to 5 µM) [ 23 , 24 ], compared with the ones used in this study (1 µM).…”

“…In this work, we aimed to disclose a pharmacological modulator to prevent BCCs extravasation by improving BBB properties and counteracting malignant cell features, ultimately hindering BCBM formation. To this end, we performed a high-throughput screening (HTS) of potential drug candidates, including several PI3K, HSP90, and EGFR inhibitors discovered based on a computer-assisted drug discovery campaign [ 39 ], and clinically approved drugs, such as fingolimod and some tetracyclines. Using an improved in vitro model of the BBB formed by confluent monolayers of mouse BMECs (b.End5) in conditions mimicking physiologic shear stress [ 40 ], we performed automated fluorescence microscopy to monitor BBB alterations upon exposure to TNBC cells (4T1), and their prevention by the selected drugs.…”

Personalized Treatment of Glioblastoma: Current State and Future Perspective

“…As previously stated, potential therapeutic options for GBM need to cross the BBB to reach therapeutically effective concentrations within the tumor region. Therefore, tools for drug discovery have been focused on candidates with special physicochemical characteristics which would permit crossing of the BBB [ 135 ]. Additionally, novel drug-delivery options, such as nanoparticles and focused ultrasound sonification, have been utilized to improve drug penetration within the tumor [ 136 , 137 ].…”