Extended Data Fig. 8: Characterization of dHTC1 and SR-1114 activity.

a, Top, a two-dimensional interaction map of dHTC1 bound to CRBN and ENL and, bottom, dHTC1 bound to ENL YEATS with key interactions highlighted. b, Left, an overlay of dHTC1 binding pose with CRBN in binary and ternary complex structures and, right, binding pose of dHTC1 with CRBN in the ternary complex with key amino acid residues highlighted. c, Peripheral CRBN loop (aa146-153) and its density (silver, threshold). d, MV4;11 ENL-TagBFP-P2A-mCherry cells (wild type or Q41A) treated with 1 µM of the indicated compounds or DMSO. BFP percentage normalized to DMSO (mean, 3 biological replicates ± S.E.M.). e, Superposition of the apo ENL YEATS domain crystal structure (6HQ0¹¹⁵, dark gray) to the dHTC1-bound ENL YEATS-CRBN/DDB1 structure showing ENL density (silver, threshold 0.03). Red arrow indicates movement of the ENL loop relative to the apo structure (aa 37-42). f, Viability of HL60 cells after 12-day treatment (cells were split 1:10 and fresh drug was added every 3 days) with (S)-dHTC1 or (R)-dHTC1 measured by ATP-dependent luminescence (normalized to DMSO, 3 independent experiments, mean of 3 biological replicates ± S.E.M.). g, Time-dependent and dose-responsive loss of HiBiT-ENL signal in MV4;11 cells treated with SR-1114. Luminescence normalized to DMSO vehicle control (mean, 4 biological replicates ± S.E.M.). h, Mean ± S.E.M. plasma concentration and pharmacokinetic properties of male C57BL/6 mice dosed with (S)-dHTC1 (25, 50 or 75 mg/kg) via intraperitoneal injection (n = 3 mice). 50 mg/kg data is plotted in Fig. 4b. i, Mean ± S.E.M of plasma concentration of male C57BL/6 mice dosed with rac-dHTC1 at 10 mg/kg via intraperitoneal (IP) injection, 2 mg/kg via intravenous (IV) injection or 2 mg/kg per os (PO) (n = 9 mice). j, Scatter plot of RNA-seq analysis of mouse-cell-depleted bone marrow lysates from MV4;11 xenograft (same experiment as data from Fig. 4e).

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