J-1063 is a potent, selective and orally bioactive ALK5 inhibitor (antagonist) with IC50 of 0.039 µM. J-1063 exerts anti-liver fibrosis effects by inhibiting inflammatory infiltration, collagen deposition, and hepatocyte necrosis. J-1063 may be used for studying liver fibrosis.

Physicochemical Properties

Molecular Formula	C24H19N5OS
Molecular Weight	425.505563020706
Exact Mass	425.131
CAS #	2374772-46-8
PubChem CID	155553747
Appearance	Typically exists as solid at room temperature
LogP	3.9
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	5
Heavy Atom Count	31
Complexity	615
Defined Atom Stereocenter Count	0
InChi Key	YVABZJGVIULAAE-UHFFFAOYSA-N
InChi Code	InChI=1S/C24H19N5OS/c1-16-6-5-9-20(26-16)24-19(22-12-17-13-25-11-10-21(17)31-22)14-29(28-24)15-23(30)27-18-7-3-2-4-8-18/h2-14H,15H2,1H3,(H,27,30)
Chemical Name	2-[3-(6-methylpyridin-2-yl)-4-thieno[3,2-c]pyridin-2-ylpyrazol-1-yl]-N-phenylacetamide
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	J-1063 (compound 4) exhibits notable inhibitory effect against ALK5 at a concentration of 10 mM (IC50 values for ALK5 and p38α MAP kinase are 8.12 and 0.039 µM, respectively) [1]. In LX-2 cells, TGF-β-induced fibroblast differentiation is improved by J-1063 (2.5, 5, 10 µM; 1 hour) [1]. J-1063 suppresses the inflammatory response generated by TGF-β at 2.5, 5, and 10 µM for one hour [1]. By controlling TGF-β/Smad signaling and preventing the activation of the NLPR3-Caspase-1 inflammasome, J-1063 prevents liver fibrosis [1]. In liver fibrosis, J-1063 (12.5 mg/kg; Cxcl1, Cxcl2 cells) prevents neutrophil and macrophage infiltration [1].
ln Vivo	J-1063 (12.5, 25, 50 mg/kg; ig, once a day for two weeks) has no harmful side effects in rats at low doses and is acceptable for therapeutic treatment [1]. J-1063 (12.5 mg/kg; orally, once daily for two weeks) was proven to be protective against TAA-induced liver fibrosis in mice [1].
Cell Assay	Western Blot analysis [1] Cell Types: LX-2 cells Tested Concentrations: 2.5, 5, 10 µM Incubation Duration: 1 h Experimental Results: Inhibition of TGF-β-induced inflammatory response.
Animal Protocol	Animal/Disease Models: Male C57BL/6 mice [1] Doses: 12.5, 25, 50 mg/kg Route of Administration: ig, one time/day, two weeks Experimental Results: Low doses have no toxic side effects on mice and are suitable for therapeutic administration . Animal/Disease Models: Male C57BL/6 mice [1] Doses: 12.5 mg/kg Route of Administration: Orally, one time/day for two weeks Experimental Results: It is beneficial to TAA-induced liver fibrosis in mice.
References	[1]. The in vitro and in vivo study of a pyrazole derivative, J-1063, as a novel anti-liver fibrosis agent: Synthesis, biological evaluation, and mechanistic analysis. Bioorg Chem. 2022; 122:105715.

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	2.3501 mL	11.7506 mL	23.5012 mL
	5 mM	0.4700 mL	2.3501 mL	4.7002 mL
	10 mM	0.2350 mL	1.1751 mL	2.3501 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.