Chlorogenic Acid Derivative B11 Inhibits Glioma Proliferation by Targeting MST4 to Regulate Oxidative Stress

ABSTRACT This study focuses on the mechanism of action of chlorogenic acid derivative B11 on glioma. Although chlorogenic acid exhibits certain anti‐tumor activity, it has limitations such as low bioavailability, broad action targets, and insufficient specificity, which restrict its practical application in tumor treatment. Therefore, chlorogenic acid derivative B11 was obtained through structural modification for investigation. Using cell experiments, animal experiments, and molecular techniques, it was found that in cell experiments, the CCK‐8 (Cell Counting Kit‐8) assay confirmed that B11 has a strong inhibitory effect on the proliferation of U118MG and LN229 glioma cells, with its IC 50 being significantly lower than that of chlorogenic acid. Growth curve and colony formation experiments further confirmed that B11 can effectively inhibit cell growth and clonogenic ability. Animal tumorigenesis experiments showed that B11 can significantly inhibit tumor growth in vivo with little impact on mouse body weight, demonstrating good safety. At the molecular mechanism level, kinase activity assays, SPR (Surface Plasmon Resonance), ITC (Isothermal Titration Calorimetry), and molecular docking techniques revealed that B11 can specifically bind to and inhibit MST4 (Mammalian Sterile 20‐like Kinase 4) kinase. RNA‐seq combined with GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analyses clarified the regulation of gene expression profiles by B11 and MST4 interference, involving pathways such as oxidative stress. Oxidative stress‐related detection found that B11 can alter mitochondrial morphology, regulate the expression of proteins such as Cyt.C (Cytochrome C) and OPA1 (Optic Atrophy 1), as well as genes such as SOD1 (Superoxide Dismutase 1), and increase ROS (Reactive Oxygen Species) levels. Immunohistochemical results showed that B11 can affect the expression of proteins such as MST4 and Nrf2 (Nuclear Factor Erythroid 2‐Related Factor 2) in tumor tissues. In conclusion, B11 can inhibit glioma cell proliferation and in vivo tumor growth by targeting MST4, regulating oxidative stress pathways, and gene expression profiles, providing a new potential drug and theoretical basis for glioma treatment.