Cat. No. : CS-7688 CAS No. : 1426138-42-2
M. Wt. : 368.82
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  • Data Sheet

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Name: FX1; 
Cat. No. : CS-7688
CAS No. : 1426138-42-2
Formula: C14H9ClN2O4S2
M. Wt. : 368.82
Solubility: DMSO: 30 mg/mL (Need ultrasonic)


FX1 is a potent and specific BCL6 inhibitor, with IC50 around 35 μM. IC50 & Target: IC50: ~35 μM (BCL6) [1] In Vitro: DLBCL cells are exposed to 50 μM FX1 for 30 minutes. FX1 profoundly reduces recruitment of BCOR and SMRT to all 3 BCL6 target genes, but not at a negative control locus. In contrast, binding of BCL6 to these same sites is not impaired, and indeed is apparently increased (perhaps because of increased accessibility to antibody). In contrast, there is little presence of SMRT at these loci in the BCL6-negative DLBCL cell line Toledo, which is not affected by FX1. The superior potency of FX1 versus 79-6 in disrupting BCL6 binding to SMRT is evident when these small molecules where compare head to head in quantitative ChIP assays in DLBCL cells after treatment with 50 μM FX1 for 6 hours. DLBCL cells are exposed to FX1 and mRNA collect at 4 serial time points. FX1 almost invariantly induce significant derepression of these genes as compare with vehicle in 2 independent DLBCL cell lines[1]. In Vivo: Spleens in FX1-treated mice are macroscopically indistinguishable from vehicle controls. As expected, total B cell abundance measured by flow cytometry is unaffected by FX1. In contrast and similar to the BCL6 BTB mutant phenotype, GC B cells (GL7+FAS+B220+) are significantly depleted by exposure to FX1. We also examine splenic architecture by IHC. Staining with B220 antibody reveal normal B cell follicular structures, whereas staining for the GC B cell–specific marker peanut agglutinin show profound loss of GCs. The half-life is estimated to be approximately 12 hours. mRNA extract from tumor tissue from these mice is assessed for abundance of BCL6 target genes CD69, CDKN1A, and CXCR4. Maximal induction of target genes occurre at 4–6 hours and subsequently decline. Finally, we assesse whether FX1 can induce toxic effects in mice. No signs of toxicity, inflammation, or infection are evident from H&E-stained sections of lung, gastrointestinal tract, heart, kidney, liver, spleen, and bone marrow of the fixed organs from mice treate with FX1 compared with vehicle. The peripheral blood counts and serum chemistry are examined in FX1-treate mice, all of which remaine within normal parameters[1].


Cell Assay: [1]Cell viability is determined with the fluorescent redox dye CellTiter-Blue. Fluorescence is determined for 3 replicates per treatment condition or vehicle with the Synergy NEO microplate reader. Cell viability of the drug-treated cells is normalized to their vehicle-treated controls, and the results are expressed as percentage viability. The drug effect as 100-percentage viability is calculated. Through dose-effect curves the drug concentration that inhibits the growth of cell lines by 50% compared with vehicle (GI50) with software is determined. Experiments are performed in triplicate. For combination treatments, cells are exposed to a dose curve of each drug alone or their combination in constant ratio, and cell viability is determined by CellTiter-Blue. To compare different schedules of treatments, we treat the cells in triplicate as follows: FX1 and doxorubicin simultaneously and cells treat for 48 hours; FX1 first and 24 hours after we add doxorubicin and treat for an extra 48 hours; doxorubicin first and 24 hours after we add FX1 and treat for an extra 48 hours. Then, the software is used to plot dose-effect curves and calculate the dose-reduction index[1]. Animal Administration: [1]Six- to 8-week-old male SCID mice are injected s.c. with 107 low-passage human SUDHL-6, OCI-Ly7, or Toledo cells. Alternatively, 6- to 8-week-old NOD/SCID mice are injected with low-passage HBL-1 cells. When tumors reach a palpable size (approximately 100 mm3), mice are assigned in a randomized way to treatment groups and treated i.p. with 25 or 50 mg/kg/d of the drugs. Drugs are reconstituted in PEG-400 and stored at -20°C until use. Tumor size is measured 3 times a week with an electronic digital caliper in 2 dimensions, and then tumor volume is calculated with the formula: tumor volume (mm3)=(smallest diameter2 × largest diameter)/2[1].


Mariano G et al. Rationally designed BCL6 inhibitors target activated B cell diffuse large B cell lymphoma. J Clin Invest. 2016 Sep 1; 126(9): 3351–3362. doi: 10.1172/JCI85795

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