XY221 (Compound 16o) selectively inhibits BRD4 BD2 with IC50 of 5.8 nM. XY221 exhibits high selectivity for pan-BD2 and BRD4 BD2 domains, 667-fold higher than BRD4 BD1 and 9-32-fold higher than BRD2/3/T BD2, respectively. XY221 induces apoptosis in MV4-11 cells and has anticancer activity.

Physicochemical Properties

Molecular Formula	C32H34FN3O5
Molecular Weight	559.63
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

Targets	BRD4 BD1 3869 nM (IC50); BRD4 BD2 5.8 nM (IC50); BRD3 BD2 70.98 nM (Ki)
ln Vitro	XY221 inhibits BRD4 BD2, BRD3 BD2, BRD2 BD2, BRDT BD2 and BRD4 BD1 with IC50 values of 5.8, 70.98, 53.47, 183, 3869 nM, respectively[1]. XY221 inhibits the proliferation of MV4−11, LNCaP, and HL-7702 cells with IC50 values of 0.51, 0.68, and 47.7 μM, respectively[1]. XY221 (500-2000 nM, 72 h) can significantly inhibit the proliferation of MV4-11 cells and induce apoptosis of MV4-11 cells[1]. The half-life of XY221 (1-10 μM, 2 h) is more than 120 min in RLM (rat liver mitochondria) and more than 120 min in HLM (human liver mitochondria) [1]. XY221 (500-2000 nM, 72h) inhibits the phosphorylation of target genes p21, ODC1, BRD4, AKT and AMPK in MV4−11 cells [1]. XY221 (2 μM, 72h) inhibits the expression of the oncogene MYC and its target gene p21 and the cell cycle-related gene BCL-2 in MV4−11 cells at the mRNA level[1].
ln Vivo	Pharmacokinetic parameters of XY221 in rats after intravenous (iv) and oral (po) administration route Cmax (μg/L) Tmax (h) AUC (0-t) (μg/L・h) AUC (0-∞) (μg/L・h) T1/2 (h) CI (L/h/kg) F (%) XY221 iv (5 mg/kg) 9790 0.03 4291 4294 1.35 1.21 XY221 po (25 mg/kg) 567 3.67 2811 2816 2.13 13.1
Cell Assay	Western Blot Analysis[1] Cell Types: MV4−11 cells Tested Tested Concentrations: 500, 2000 nM Incubation Duration: 24-72 h Experimental Results: Inhibited target genes p21 ,ODC1, BRD4 and the phosphorylation of AKT and AMPK. Real Time qPCR[1] Cell Types: MV4−11 cells Tested Tested Concentrations: 2 μM Incubation Duration: 72 h Experimental Results: Inhibited target genes p21 ,ODC1, BRD4 and the phosphorylation of AKT and AMPK. Suppressed oncogene MYC and its target gene p21 and cell-cycle-related gene BCL-2 at mRNA level.
References	[1]. Discovery of novel BRD4-BD2 inhibitors via in silico approaches: QSAR techniques, molecular docking, and molecular dynamics simulations. Mol Divers. 2024 Apr;28(2):671-692.

Solubility Data

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.7869 mL	8.9345 mL	17.8689 mL
	5 mM	0.3574 mL	1.7869 mL	3.5738 mL
	10 mM	0.1787 mL	0.8934 mL	1.7869 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.