A-485 has a robust antileukemic effect on primary AML cells. (A) The effect of A-485 on the growth of human MLLr AML cell lines. Cell lines were exposed to A-485 at 1 μM for 48 hours (n = 3; mean ± SD). (B) Cell-cycle profile in THP-1 with A-485 (1uM) treatment for 48 hours (n = 3; mean ± SD). (C) Surface expression of CD11b and CD11c in THP-1 cells with A-485 (1 μM) treatment for 48 hours. (D) HOXA9 and MEIS1 expression in THP-1 cells treated with A-485 (1 μM) treatment for 48 hours (n = 3; mean ± SD). (E) Representative bioluminescent imaging of NSG recipient mice that underwent transplantation with THP-1 cells at indicated A-485 treatment time. (F) Frequencies of leukemia cells (hCD45⁺) in the peripheral blood. (n = 5; mean ± SD). (G) Kaplan-Meier plot of A-485– and vehicle-treated mice that underwent transplantation with THP-1 cells. P value was generated using Mantel-Cox log-rank test. (H) Dose dependent effect of A-485 inhibitor on CRISPR-engineered acute leukemias bearing MLL-AF9 translocation. Cells were exposed to the indicated concentrations of A-485 for 48 hours (n = 3; mean ± SD). (I) Clonogenic assays with primary MLLr AML cells from patients treated with A-485. (n = 8; mean ± SD). ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001.