Fig. 1
Structure and in vitro activity of the Menin-MLL1 inhibitor DS-1594a·succinate. A Chemical structure of DS-1594a·succinate (succinic acid salt of DS-1594a). B Overall structure of the DS-1594a·succinate/human Menin complex showing 4 chains of Menin included in 1 asymmetric unit (left) and Fo-Fc electron density map of DS-1594a·succinate bound to chain A of Menin contoured at 3 sigma (right). C Inhibition of the Menin-MLL1 interaction by DS-1594a·succinate in a cell-free assay (AlphaLISA assay). The inhibition (%) of the Menin-MLL1 interaction by DS-1594a·succinate is shown as the mean ± SD at each concentration of DS-1594a·succinate (1000, 170, 28, 4.6, 0.77, 0.13, 0.021, and 0.0036 nM; n = 4). D Inhibition of the Menin-MLL1 interaction by DS-1594a·succinate in a cellular assay. Co-immunoprecipitation (Co-IP) was performed with an anti-HA antibody or control IgG in human embryonic kidney 293 T (HEK293T) cells transfected with HA-MLL1N1-1395 upon treatment with DMSO (0) or DS-1594a·succinate (10, 30, 100, 300, 1000 nM). The input represents lysate samples before Co-IP treatment. E In vitro growth-inhibitory effects of DS-1594a·HCl (the HCl salt of DS-1594a) against leukemia cell lines (left). Titration curves from the cell viability assay (CellTiter-Glo) were created after 7 days of treatment of human leukemic cell lines with DS-1594a·HCl. The horizontal axis is the common logarithm of the concentration of DS-1594a·HCl, and the mean (± SD) survival rates at each concentration are shown (n = 3). The GI50 concentrations of DS-1594a·HCl for each cell line are shown (right). B-ALL, B-cell acute lymphoblastic leukemia; CML, chronic myeloid leukemia; APL, acute promyelocytic leukemia; Co-IP, coimmunoprecipitation