AP-III-a4 (ENOblock) hydrochloride is a nonsubstrate analogue enolase inhibitor with an IC 50 of 0.576 uM. AP-III-a4 hydrochloride can be used for the research of cancer and diabetic In Vitro AP-III-a4 (ENOblock) (0-10 μM; 24 h) inhibits HCT116 cell viability in a dose-dependent manner. AP-III-a4 directly binds to enolase and inhibits its activity. AP-III-a4 (0-10 μM; 24 or 48 h) inhibits cancer cell migration and invasion, induces cancer cell apoptosis. AP-III-a4 (10 μM; 24 h) can induce glucose uptake and inhibit phosphoenolpyruvate carboxykinase (PEPCK) expression in hepatocytes and kidney cells. MCE has not independently confirmed the accuracy of these methods. They are for reference only. Cell Viability AssayCell Line: HCT116 Concentration: 1.25, 2.5, 5 and 10 μM Incubation Time: 24 h Result: Induced higher levels of HCT116 colon cancer cell death in hypoxic conditions compared to normoxia. Western Blot AnalysisCell Line: HCT116 Concentration: 1.25, 2.5, 5 and 10 μM Incubation Time: 24 h for AKT, 48 h for Bcl-Xl Result: Bound to enolase in cell lysate and bound to purified enolase. Decreased the expression of AKT and Bcl-Xl, which are negative regulators of apoptosis. Cell Invasion AssayCell Line: HCT116 Concentration: 0.156, 0.312, 0.625, 1.25 and 2.5 μM Incubation Time: 24 h Result: Significantly inhibits cancer cell invasion at a treatment concentration of 0.625 μM. Cell Migration AssayCell Line: HCT116 Concentration: 0.625, 1.25 and 2.5 μM Incubation Time: 24 h Result: Inhibited cell migration dose-dependently. RT-PCRCell Line: Huh7 and HEK Concentration: 10 μM Incubation Time: 24 h Result: Induced glucose uptake and inhibited PEPCK expression. In Vivo AP-III-a4 (ENOblock) (10 μM; 96 h) inhibits cancer cell metastasis and suppresses the gluconeogenesis regulator PEPCK in zebrafish . MCE has not independently confirmed the accuracy of these methods. They are for reference only. Animal Model: The zebrafish cancer cell HCT116 xenograft model Dosage: 10 μM Administration: 96 h Result: Reduced cancer cell dissemination. Inhibited PEPCK expression and induced glucose uptake. Inhibited adipogenesis and foam cell formation. Form:Solid IC50& Target:IC 50 : 0.576 uM (enolase)