Discovery, Optimization, and Biological Evaluation of 2‑Cyano-2-(9H‑xanthen-9-ylidene)acetamide Derivatives as ZNF207 Inhibitors for Anti-Glioma Therapy

Zinc finger protein 207 (ZNF207) is highly expressed in glioma and represents a promising therapeutic target. Building on the reported inhibitor C16, we developed a novel derivative, TMLZ-G1, with improved stability and a removed chiral center. Subsequent optimization identified TMLZ-G46, which showed high affinity for ZNF207 (KD = 68 nM), potent antiproliferative activity against ZNF207-high glioma cells (IC50 = 0.93–2.07 μM), and strong inhibition of stemness (IC50 = 0.34–0.58 μM). TMLZ-G46 suppressed colony formation, migration, and invasion; induced cell cycle arrest and apoptosis; and displayed favorable pharmacokinetics with 68.1% oral bioavailability, brain penetration, and no P-glycoprotein efflux. In vivo, TMLZ-G46 achieved 83.7% tumor growth inhibition in subcutaneous glioblastoma (GBM) (grade IV) cell line-derived xenografts without detectable toxicity and significantly prolonged survival in an orthotopic GBM xenograft model, with efficacy comparable to temozolomide. These findings highlight TMLZ-G46 as a promising candidate for glioma therapy.