HIF-1/2α-IN-2 is a potent inhibitor of HIF-1/2α that effectively reduces HIF-1/2α levels and elicits an iron starvation response by specifically targeting ISCA2, a key protein in Iron-Sulfur Cluster Assembly 2. [1]

HIF-1/2α-IN-2 (compound #1), at concentrations of 0 to 25 μM over 24 hours, primarily reduces HIF-2α translation rather than transcription, leading to decreased VEGFA and POU5F1 transcription without significantly affecting EPAS1 (HIF-2α) [1]. At 0 to 100 μM, the same compound inhibits luciferase production in an Iron-Responsive Element (IRE) luciferase reporter driven by HIF-2α, with an IC 50 value of 3.9 μM, thus effectively blocking IRE-dependent HIF-2α translation [1]. Additionally, at 0, 10, and 50 μM, HIF-1/2α-IN-2 protects ISCA2 from Pronase-mediated degradation at 4 μg/mL, and at 1.5 μM, it causes accumulation of iron and metals in normoxia 786-0 cells by targeting ISCA2 [1]. At concentrations up to 100 μM over 24 hours, it inhibits ISCA2 and induces ferroptosis [1]. Western blot analysis with normoxia 786-0 cells and hypoxic ACHN cells, exposed to 0, 1, 5, 10, and 25 μM for 24 hours, reveals decreased protein levels of HIF-2α, FTH1, ISCA2, and increased IRP2 protein levels at concentrations over 10 μM in 786-0 cells, and decreased HIF-2α and FTH1 levels at 1 μM in ACHN cells [1]. Cell viability assays indicate that HIF-1/2α-IN-2 inhibits 786-0 cell viability by inducing ferroptosis, with an IC 50 value of 22.0 μM [1].