HDAC-IN-50 is a potent, orally bioactive dual (bifunctional) inhibitor of FGFR and HDAC, with IC50s of 0.18, 1.2, 0.46, and 1.4 for FGFR1, FGFR2, FGFR3, FGFR4, HDAC1, HDAC2, HDAC6, and HDAC8 respectively. , 1.3, 1.6, 2.6, 13 nM. HDAC-IN-50 causes apoptosis (Apoptosis) and cell cycle arrest in the G0/G1 phase. HDAC-IN-50 reduces the expression of pFGFR1, pERK, and pSTAT3. HDAC-IN-50 has anticancer effect.

Physicochemical Properties

Molecular Formula	C31H41N7O4
Molecular Weight	575.701746702194
Exact Mass	575.322
CAS #	2653339-26-3
PubChem CID	166642476
Appearance	Typically exists as solid at room temperature
LogP	3.5
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	16
Heavy Atom Count	42
Complexity	780
Defined Atom Stereocenter Count	0
InChi Key	WWGLOQFMJUZFTJ-UHFFFAOYSA-N
InChi Code	InChI=1S/C31H41N7O4/c1-22(2)32-12-14-38(25-15-26(41-3)18-27(16-25)42-4)24-10-11-28-29(17-24)35-30(20-33-28)23-19-34-37(21-23)13-8-6-5-7-9-31(39)36-40/h10-11,15-22,32,40H,5-9,12-14H2,1-4H3,(H,36,39)
Chemical Name	7-[4-[7-[3,5-dimethoxy-N-[2-(propan-2-ylamino)ethyl]anilino]quinoxalin-2-yl]pyrazol-1-yl]-N-hydroxyheptanamide
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	FGFR1 0.18 nM (IC50) FGFR2 1.2 nM (IC50) FGFR3 0.46 nM (IC50) FGFR4 1.4 nM (IC50) HDAC1 1.3 nM (IC50) HDAC2 1.6 nM (IC50) HDAC6 2.6 nM (IC50) HDAC8 13 nM (IC50)
ln Vitro	Compound 10e, HDAC-IN-50 (0.1, 1, 10, 100 nM; 12-84 h) promotes time- and dose-dependent apoptosis and cell cycle arrest in the G0/G1 phase[1]< /sup >. In a dose-dependent manner, HDAC-IN-50 (0, 1.25, 2.5, 5 µM for HCT116 cells, 0, 1, 10, 100 nM for SNU-16 cells; 36 h) decreases the expression of pFGFR1, pERK, and pSTAT3.
ln Vivo	Mice treated orally with HDAC-IN-50 (15, 30 mg/kg; once daily for 18 days) exhibit anti-tumor action [1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: HCT116, SNU-16, KATO III, A2780, K562, Jurkat cells Tested Concentrations: 0-30 µM Incubation Duration: 72 h Experimental Results: demonstrated antiproliferative activities with IC50s of 0.82, 0.0007, 0.0008, 0.04, 2.46, 15.14 µM for HCT116, SNU-16, KATO III, A2780, K562, Jurkat cells, respectively. Cell Cycle Analysis[1] Cell Types: SNU-16 cells Tested Concentrations: 0.1, 1, 10, 100 nM Incubation Duration: 12, 24, 36 h Experimental Results: Induced cell cycle arrest at G0/G1 phase in a time and dose-dependent manner. Apoptosis Analysis[1] Cell Types: SNU-16 cells Tested Concentrations: 0.1, 1, 10, 100 nM Incubation Duration: 36, 48, 60, 72, 84 h Experimental Results: Induced apoptosis with the apoptotic rate increased 30.8% and 49.6% at 10, 100 nM, respectively. Western Blot Analysis[1] Cell Types: HCT116, SNU-16 cells Tested Concentrations: 0, 1.25, 2.5, 5 µM for HCT116 cells, 0, 1, 10, 100 nM for SNU-16 cells Incubation Duration: 36 h Experimental Results: decreased the expression of pFGFR1, pERK, pSTAT3 in a dose-dependent manner.
Animal Protocol	Animal/Disease Models: BALB/c nude mice (HCT116 xenograft model)[1] Doses: 15, 30 mg/kg Route of Administration: Po; daily for 18 days Experimental Results: Inhibited the tumor growth and downregulated the expression of pSTAT3, pFGFR1, increased the expression of Ac-H3.
References	[1]. Design and Synthesis of Fibroblast Growth Factor Receptor (FGFR) and Histone Deacetylase (HDAC) Dual Inhibitors for the Treatment of Cancer. J Med Chem. 2022 Dec 22;65(24):16541-16569.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.7370 mL	8.6851 mL	17.3702 mL
	5 mM	0.3474 mL	1.7370 mL	3.4740 mL
	10 mM	0.1737 mL	0.8685 mL	1.7370 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.