KC7F2 is a potent HIF-1 pathway inhibitor with potential anti-cancer activity.

HIF1α:20 μM

KC7F2在LN229-HRE-AP细胞中抑制HRE驱动的转录并降低HIF-1α蛋白水平。对于MCF7、LNZ308、A549、U251 mg和LN229癌细胞，KC7F2表现出剂量响应性细胞毒性，IC50约为15至25μM。在D54 mg胶质瘤细胞中，尤其在缺氧条件下，KC7F2抑制了集落形成。[1] 在缺氧的小胶质细胞培养中，KC7F2下调TfR和DMT的表达，并减少了HIF-1α介导的铁积累。[2]

KC7F2在戊四氮癫痫模型大鼠中显著缩短潜伏期，并在慢性阶段增加了锂-毛果芸香碱大鼠模型中自发性复发性癫痫的发作率。[3]

HIF transcriptional activity assay: Cells are incubated at 37 in a humidified atmosphere containing 5% CO2 and 21% O2 (normoxia) or 1% O2 (hypoxia) in a hypoxia workstation. The LN229-HRE-AP reporter cell line for HIF transcriptional activity is created by stably transfecting LN229 cells with the pACN188 plasmid, which contains an alkaline phosphatase gene driven by six HREs derived from the VEGF gene.

Cells are seeded onto 96-well plates (4 × 103/well) and cultured under normoxic (21% O2) and hypoxic (1% O2) conditions with different concentrations of KC7F2 for 72 h or treated for various times with 20 μM KC7F2. For proliferation analysis, cells are fixed with 50% trichloroacetic acid for 1 h at 4°C, followed by staining with 0.4% sulforhodamine B dissolved in 1% acetic acid for 30 min at room temperature. Plates are washed five times with 1% acetic acid to remove unbound dye. Bound dye is dissolved by adding 10 mM unbuffered Tris base. Cell proliferation is calculated by measuring OD values at 564 nm using a spectrophotometer.(Only for Reference)