BRD4/NAMPT-IN-1 (Compound A2) has a strong inhibitory effect on NAMPT and BRD4 (IC50=35 nM (NAMPT) and 58 nM (BRD4)). BRD4/NAMPT-IN-1 significantly inhibits the growth and migration of liver cancer cells and promotes cell apoptosis. BRD4/NAMPT-IN-1 also exhibited potent anticancer effects in HCCLM3 xenograft mouse models without obvious toxic effects.

Physicochemical Properties

Molecular Formula	C30H30CLN7O2S
Molecular Weight	588.12
Appearance	Solid powder
HS Tariff Code	2934.99.9001
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(BD1BD2) 58 nM (IC50) BRD4 (BD1) 12 nM (IC50) BRD4 (BD2) 41 nM (IC50)
ln Vitro	The IC50 of BRD4/NAMPT-IN-1 against other members of the BET family is 12 nM (BRD4(BD1)) and 41 nM (BRD4(BD2))[1]. The IC50 of BRD4/NAMPT-IN-1 in inhibiting cancer cell proliferation is 2.37 μM (Hep3B), 6.49 μM (Huh7), 5.44 μM (HCCLM3) and 9.51 μM (LX-2)[1]. Treatment of Hep3B cells with BRD4/NAMPT-IN-1 (1-10 μM; 72 h) revealed that: it can inhibit the expression of oncogenes upregulated by BRD4, while reducing the levels of NAPRT and NAMPT; significantly increase the arrest of cells in the G0/G1 phase; induce cell apoptosis in a dose-dependent manner; and inhibit cell migration in a dose-dependent manner[1]. BRD4/NAMPT-IN-1 (1-10 μM; 72 h) dose-dependently reduces the concentration of NAD + in Hep3B cells and HCCLM3 cells [1]. Apoptosis Analysis[1] Cell Line: Hep3B cells Concentration: 1; 5; 10 μM Incubation Time: 72 h Result: The apoptosis rate induced was significantly higher than that of the control FK866 (HY-50876) and JQ1 (HY-13030) at the same dose. Cell Cycle Analysis[1] Cell Line: Hep3B cells Concentration: 1; 5; 10 μM Incubation Time : 72 h Result: Significantly increased the accumulation of Hep3B cells at the G0/G1 stage over the commonly used hepatocellular carcinoma therapeutic agents FK866 (HY-50876) and JQ1 (HY-13030).
ln Vivo	BRD4/NAMPT-IN-1 (ip; 40 mg/kg/day and 80 mg/kg/day; for 27 days) exhibited a dose-dependent tumor inhibitory effect in nude mice bearing HCCLM3 xenografts [1].
Animal Protocol	Animal/Disease Models:HCCLM3 xenograft nude mice [1] Doses: 40 mg/kg/day and 80 mg/kg/day Route of Administration: i.p.; 27day Experimental Results: Inhibited the growth of HCCLM3 tumors significantly in both groups at two different doses, with significant decreases in tumor volume and weight. In the 40 mg/kg dose group, the tumor growth inhibition rate reached 37.20%, and in the 80 mg/kg dose group, the tumor growth inhibition rate reached 58.17%. Showed no significant weight loss or other significant toxic side effects.
References	[1]. Discovery of potent and novel dual NAMPT/BRD4 inhibitors for efficient treatment of hepatocellular carcinoma. Eur J Med Chem. 2024 May 5;271:116444.

Solubility Data

Solubility (In Vitro)

DMSO : 100 mg/mL (170.03 mM; with sonication)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.25 mM)(Saturation unknown) in 10% DMSO 40% PEG300 5% Tween-80 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution, add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix well; then add 50 μL Tween-80 to the above system and mix well; then continue to add 450 μL saline to make up to 1 mL. *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (4.25 mM)(Saturation unknown) in 10% DMSO 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution, add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD in saline and mix well. *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 3: ≥ 2.5 mg/mL (4.25 mM)(Saturation unknown) in 10% DMSO 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution, add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL corn oil and mix well.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.7003 mL	8.5017 mL	17.0033 mL
	5 mM	0.3401 mL	1.7003 mL	3.4007 mL
	10 mM	0.1700 mL	0.8502 mL	1.7003 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.