ZXH-3-26

ZXH-3-26 is a CRBN-recruiting BRD4 PROTAC degrader with a DC_50/5h of approximately 5 nM. ZXH-3-26 drives the ubiquitination and degradation of BRD4 by recruiting BRD4_BD1 to the CRL4^CRBN E3 ubiquitin ligase complex, thereby inhibiting the dynamic distribution of super-enhancers. ZXH-3-26 reverses TNF-α-induced impairment of the immunosuppressive function of mesenchymal stem cells, and affects RNA synthesis and the transcriptional elongation process of Pol II. ZXH-3-26 can be used in studies related to inflammatory arthritis, malignant tumors, and HIV latency^{[1][2][3][4][5][6][7]}. In Vitro:ZXH-3-26 restores the immunosuppressive function of human bone marrow mesenchymal stem cells (MSCs) pretreated with TNF-α, and reduces the proliferation level of CD³⁺ T cells to a level comparable to that of untreated MSCs^[6].
ZXH-3-26 (48 h) reverses the TNF-α-induced upregulation of LIF mRNA, intracellular protein, and secreted protein levels in human bone marrow mesenchymal stem cells (MSCs)^[6].
ZXH-3-26 (100 nM; 3 h) efficiently depletes BRD4 in Tir1+ WT26 mouse embryonic cells expressing the BRD2 degrader, triggering severe defects in RNA synthesis and Pol II pausing near promoters; when used in combination with auxin to co-deplete BRD2, it further inhibits RNA synthesis, reverses the accumulation of paused Pol II, and reduces transcription initiation^[7].
ZXH-3-26 (1 nM-10 μM; 5 h) potently degrades BRD4BD1 in Flip-In 293 reporter cells, with a DC_50/5h value of approximately 5 nM; moreover, it exhibits no detectable activity against BRD2BD1, BRD3BD1 and BRD4BD2 at concentrations > 10 μM^[2].
ZXH-3-26 (1 nM-10 μM; 5 h) effectively degrades endogenous full-length BRD4 in HEK293T cells without affecting the levels of endogenous BRD2 and BRD3 ^[2].
ZXH-3-26 (0.1 μM; 5 h) selectively downregulates BRD4 in MM.1s cells after treatment at a concentration of 0.1 μM for 5 h, while exerting no significant effects on other proteins in the proteome^[2].
ZXH-3-26 (0.001-10 μM; 0.5-24 h) degrades both BRD4 isoforms in HeLa cells in a dose- and time-dependent manner, achieving complete depletion of BRD4 after 4 h of treatment at a concentration of 100 nM^[3].
ZXH-3-26 (100 nM; 2 h) reduces the number of BRD4 condensates and the fluorescence intensity of EGFP-BRD4 in HeLa cells expressing EGFP-BRD4^[3].
After washout of ZXH-3-26 (100 nM; 6 h treatment followed by 18 h recovery), BRD4 condensates in HeLa cells preferentially and fully recover within 18 h, despite only partial restoration of BRD4 protein levels^[3].
ZXH-3-26 (100 nM; 6 h treatment, 18-42 h wash-out) reduces the BRD4 binding level at SE regions in HeLa cells; after wash-out, BRD4 condensates preferentially recover at SE regions, which is correlated with the restoration of gene expression to the levels observed in the control group^[3].
ZXH-3-26 (100 nM; 6 h) reduces the number of CYCT1 and MED1 condensates in HeLa cells^[3].
ZXH-3-26 (0.05 μM; 24 h) disrupts the BRD4-CyclinT1 interaction, but does not induce the upregulation of HEXIM1 protein in Jurkat cells treated with 0.05 μM for 24 h^[4].
ZXH-3-26 (0.01-0.1 μM; 24 h) does not induce the upregulation of HEXIM1 protein in JLatA2 cells^[4].
ZXH-3-26 (0.05-0.25 μM; 24 h) does not induce HEXIM1 promoter-driven luciferase activity in 293T-HEXIMpr104 cells, and only a weak inductive effect is observed after incubation for 24 h at the non-selective concentration of 0.25 μM^[4].
ZXH-3-26 (0.001-0.1 μM; 8-24 h) efficiently and selectively degrades BRD4L in primary CD4⁺ T cells from healthy donors at a concentration of 0.005 μM, while more extensive degradation of the BET family occurs at concentrations ≥ 0.05 μM^[4].
ZXH-3-26 (0.01 μM; 24 h) efficiently degrades BRD4L at a concentration of 0.01 μM, but does not induce upregulation of HEXIM1 protein after 24 h of incubation with resting and total primary CD4⁺ T cells from healthy donors^[4].
ZXH-3-26 (0.005 μM; 24 h) at the BRD4-selective concentration of 0.005 μM fails to induce HIV latency reversal in primary CD4⁺ T cells derived from ART-suppressed donors, either when used alone or in combination with AZD5582 (HY-12600) ^[4].
ZXH-3-26 (0.001-20 μM; 24 h) does not induce HIV latency reversal in JLatA2 cells at BRD4-selective concentrations (0.005-0.05 μM), and GFP induction occurs only at higher non-selective concentrations^[4].
ZXH-3-26 (0.001-20 μM; 24 h) does not induce HIV latency reversal in JLat10.6 cells at any of the tested concentrations^[4].
ZXH-3-26 (0.005-0.5 μM; 24 h) does not exhibit synergistic latency-reversing activity against HIV at any of the tested concentration combinations when used in combination with AZD5582 in JLat10.6 cells^[4].