Fig. 4: HTC affords a stereoselective degrader with enhanced in vivo properties.
From: High-throughput ligand diversification to discover chemical inducers of proximity
a, Viability of MV4;11 cells after 12-day treatments with (S)-dHTC1 or (R)-dHTC1. Cells were split 1:10 and fresh drug was added every 3 days; then, viability was measured by ATP-dependent luminescence (normalized to DMSO; n = 3 independent experiments; mean ± s.e.m.; n = 3 biological replicates). b, Mean plasma concentration of (S)-dHTC1 and SR-1114 in male C57BL/6 mice dosed with 50 mg kg−1 through intraperitoneal injection (n = 3 mice). Cellular DC50 (MV4;11 at 8 h) is depicted with and without adjustment for PPB in both plots. (S)-dHTC1 data are repeated in Extended Data Fig. 8h. c, Diagram representing the timeline of dosing and data collection for in vivo studies. d, Immunoblot analysis of mouse-cell-depleted bone marrow from MV4;11 xenotransplantation following treatment with vehicle, (S)-dHTC1 (50 mg kg−1) or (R)-dHTC1 (50 mg kg−1) through intraperitoneal injection (n = 4 mice per group). e, RNA-seq analysis of mouse-cell-depleted bone marrow lysates from MV4;11 xenograft (n = 3 individual mice per treatment condition from xenograft experiment shown in c). P values were calculated using DESeq2 (two-sided Wald’s test). Data are reproduced in Extended Data Fig. 8j. f, Box-and-whisker plot depicting the effect of (S)-dHTC1 on the mRNA expression of ENL target genes compared to vehicle control. ENL target genes, defined by typical (gray; n = 176 genes) and asymmetric (blue; n = 56 genes) ENL ChIP-seq signal strength within the gene promoter region (typical and asymmetric target genes) or by responsiveness to ENL degradation through dTAG-enabled chemically induced degradation (green; n = 150 genes). Nontargets include all other expressed transcripts (n = 14,854 genes). Center lines indicate the median, boxes indicate the first to third quartiles and whiskers represent 1.5× the interquartile range. g, Flow-cytometry-based quantification of hCD11b expression in indicated tissues (n = 3 mice; same mice as in d). Statistical analysis was conducted using an ordinary two-way analysis of variance with uncorrected Fisher’s least significant difference multiple-comparisons test (two-sided). *P < 0.05 and ****P < 0.0001 (for blood, P = 0.0182; for spleen, P < 0.0001.)
