KZL204, a Trifluoromethylated Quinazoline Derivative, Exhibits High Potent Anti-Tumor Effects on Glioblastoma Multiforme U251MG Cells.

Abstract

Objective: Recently, there has been much interest in quinazoline derivatives due to their unique anti-tumor effects. In this study, we aimed to investigate the effects of KZL204, an active trifluoromethylated quinazoline derivative, on a human glioblastoma multiforme (GBM) cell line U251MG. Additionally, we tried to identify the potential target of KZL204 for treating GBM.

Methods: Cell counting kit-8 (CCK-8) assay for cytotoxicity, 5-ethynyl-2-deoxyuridine (EdU) staining for cell proliferation, flow cytometry for cell apoptosis and cell cycle, wound scratch test for cell migration, and transwell assay for cell invasion were carried out on U251MG cells after exposing them to different concentrations of KZL204. In addition, western blot analysis, network pharmacology-based analysis, molecular docking assay, cellular thermal shift assay (CETSA), and cycloheximide chase assay were performed.

Results: Our results showed that KZL204 concentration-dependently inhibited U251MG cell proliferation, induced apoptosis, arrested cell cycle in the G2/M phase, and inhibited cell invasion and migration capacity. Further network pharmacology-based analysis revealed that epidermal growth factor receptor (EGFR), FYN, YES1, LYN, ephrin type-A receptor 2 (EPHA2), and EPHA4 are the top 6 core targets for inhibiting cell growth, apoptosis, cell cycle, and metastasis of the GBM cells. Molecular docking and CETSA showed that KZL204 had a strong targeting binding affinity with EPHA2. Cycloheximide chase assay and western blot results demonstrated that KZL204 could down-regulate the protein level of EPHA2.

Conclusions: KZL204 exhibits potent inhibitory activity for glioblastoma multiforme cells, which may be related to its role in promoting the degradation of EPHA2.