LCL-461 has anti-AML effect in AML mouse model and in human AML blasts. (A) Measured weights of harvested spleen at day 30 after injection of cells. (B) Measured weights of harvested liver. (C-D) FACS analysis to measure the percentage of injected MV4-11-R cells (hCD45⁺ cells) in the bone marrow. Values indicate mean ± SD of n = 6 mice. P values are generated using ANOVA. (E) MS measuring LCL-461 in sorted hCD45⁺ cells vs hCD45⁻ cells. (F) Western blot to measure ACO2 and LC3B in sorted hCD45⁺ cells. (G) Electron microscopy visualization of the morphology of sorted hCD45⁺ cells obtained from bone marrow of mice injected with vehicle or LCL-461. Values indicate mean ± SD of n = 3 mice. P values are generated using the 2-sided Student t test. (H) AML bone marrow blasts obtained from FLT3⁺ patients were treated with different doses of LCL-461 to measure percentage of viability at 24 hours. (I) AML bone marrow blasts obtained from FLT3⁺ patients were treated with 8 μM LCL-461 followed by western blot to measure ACO2 matrix protein degradation and LC3B lipidation. (J) Scheme illustrating the mechanism by which targeting mitochondria using LCL-461 results in lethal mitophagy in FLT3⁺ AML.