HJC0416 is a potent orally bioactive STAT3 inhibitor. HJC0416 displays anti-proliferation activity and causes apoptosis. HJC0416 reduces the expression of p-STAT3 (Tyr-705), Cyclin D1 and increases the expression of cleaved caspase-3 protein. . HJC0416 displays anti-tumor effects.

Physicochemical Properties

Molecular Formula	C18H17CLN2O4S
Molecular Weight	392.856582403183
Exact Mass	392.059
CAS #	1617518-22-5
PubChem CID	77105973
Appearance	Typically exists as solid at room temperature
LogP	2.1
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	6
Heavy Atom Count	26
Complexity	634
Defined Atom Stereocenter Count	0
InChi Key	AUTAJMGNRLHARZ-UHFFFAOYSA-N
InChi Code	InChI=1S/C18H17ClN2O4S/c19-13-3-5-16(25-8-1-7-20)15(10-13)18(22)21-14-4-2-12-6-9-26(23,24)17(12)11-14/h2-6,9-11H,1,7-8,20H2,(H,21,22)
Chemical Name	2-(3-aminopropoxy)-5-chloro-N-(1,1-dioxo-1-benzothiophen-6-yl)benzamide
Synonyms	HJC 0416; HJC-0416; HJC0416
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

ln Vitro	In MDA-MB-231 cells, HJC0416 (0-10 µM; 48 h) has anti-proliferative activity and causes cellular inflammation [1]. p-STAT3 is decreased by HJC0416 (0-10 µM; 12 h) (Tyr-705).
ln Vivo	HJC0416 (10 mg/kg intraperitoneally and 100 mg/kg intravenously, once a day for seven days) decreased tumor growth in mice without a discernible drop in body weight [1].
Cell Assay	cell proliferation assay [1] Cell Types: MCF-7, MDA-MB-231, AsPC1, Panc-1 Cell Tested Concentrations: 0-100 µM culture, expression of Cyclin D1 and expression of cleaved caspase-3 protein [1]. Incubation Duration: 72 hrs (hours) Experimental Results: demonstrated anti-proliferative activity against MCF-7, MDA-MB-231, AsPC1, and Panc-1 cells, with IC50s of 1.76, 1.97, 0.04, and 1.88 µM, respectively. Apoptosis analysis[1] Cell Types: MDA-MB-231 Cell Tested Concentrations: 1, 5, 10 µM Incubation Duration: 48 hrs (hours) Experimental Results: Induced apoptosis. Western Blot Analysis[1] Cell Types: MDA-MB-231 Cell Tested Concentrations: 0, 1, 5, 10 µM Incubation Duration: 12 hrs (hours) Experimental Results: Inhibition of the expression of phosphorylated STAT3 at Tyr-705 and Cyclin D1, and increased cleaved caspase -3.
Animal Protocol	Animal/Disease Models: 6 weeks, female nude mice (MDA) -MB-231 cells) [1] Doses: 10 mg/kg intraperitoneally (ip) (ip); 100 mg/kg orally. Route of Administration: intraperitoneal (ip) injection or oral administration; one time/day for 7 days Experimental Results: Compared with intraperitoneally (ip) (ip) injected control mice, tumor volume was diminished by 67%; at a dose of 100 mg/kg, mouse xenograft tumor growth was also Dramatically diminished by 46%.
References	[1]. Discovery of potent anticancer agent HJC0416, an orally bioavailable small molecule inhibitor of signal transducer and activator of transcription 3 (STAT3). Eur J Med Chem. 2014 Jul 23;82:195-203.

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	2.5454 mL	12.7272 mL	25.4544 mL
	5 mM	0.5091 mL	2.5454 mL	5.0909 mL
	10 mM	0.2545 mL	1.2727 mL	2.5454 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.