Discovery, Optimization, and Biological Evaluation of 2-Cyano-2-(9 <i>H</i> -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 <b>C16</b>, we developed a novel derivative, <b>TMLZ-G1</b>, with improved stability and a removed chiral center. Subsequent optimization identified <b>TMLZ-G46</b>, which showed high affinity for ZNF207 (<i>K</i>_D = 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). <b>TMLZ-G46</b> 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, <b>TMLZ-G46</b> 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 <b>TMLZ-G46</b> as a promising candidate for glioma therapy.