A 77-01 is a potent inhibitor of the TGF-β type I receptor family ALK5 with IC50 of 25 nM.

Physicochemical Properties

Molecular Formula	C18H14N4
Molecular Weight	286.33
Exact Mass	286.122
CAS #	607737-87-1
PubChem CID	10469233
Appearance	Off-white to light yellow solid powder
Density	1.254g/cm3
Boiling Point	481.63ºC at 760 mmHg
Flash Point	256.391ºC
Index of Refraction	1.685
LogP	3.995
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	3
Rotatable Bond Count	2
Heavy Atom Count	22
Complexity	374
Defined Atom Stereocenter Count	0
InChi Key	KJTYZDORHCDZPS-UHFFFAOYSA-N
InChi Code	InChI=1S/C18H14N4/c1-12-5-4-8-17(21-12)18-15(11-20-22-18)13-9-10-19-16-7-3-2-6-14(13)16/h2-11H,1H3,(H,20,22)
Chemical Name	4-[5-(6-methylpyridin-2-yl)-1H-pyrazol-4-yl]quinoline
Synonyms	A77-01 A-77-01 A 77-01
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	Treatment with A-77-01 (0.01-10 µM; 25 hours; Mv1Lu cells) potently suppresses the concentration-dependent transcriptional activation elicited by TGF-β [1]. The growth-inhibitory effects of TGF-β are successfully prevented when Mv1Lu cells are treated with A-77-01 (1 µM) for 24-72 hours [1]. TGF-β-induced Smad2 phosphorylation is inhibited when HaCaT cells are treated with A-77-01 (1 µM) for 30 minutes [1].
Cell Assay	RT-PCR[1] Cell Types: Mv1Lu Cell Tested Concentrations: 0.01 µM, 0.03 µM, 0.1 µM, 0.3 µM, 1 µM, 3 µM, 10 µM Incubation Duration: 25 hrs (hours) Experimental Results: Inhibition of TGF-β-induced luciferase activity Concentration-dependent fashion. Cell viability assay [1] Cell Types: Mv1Lu Cell Tested Concentrations: 1 µM Incubation Duration: 24 hrs (hours), 48 hrs (hours), 72 hrs (hours) Experimental Results: Effectively prevents the growth inhibitory effect of TGF-β. Western Blot Analysis[1] Cell Types: HaCaT cells Tested Concentrations: 1 µM Incubation Duration: 30 minutes Experimental Results: Inhibition of TGF-β-induced Smad2 phosphorylation.
References	[1]. The ALK-5 inhibitor A-83-01 inhibits Smad signaling and epithelial-to-mesenchymal transition by transforming growth factor-beta. Cancer Sci. 2005 Nov;96(11):791-800.

Solubility Data

Solubility (In Vitro)

DMSO : ~25 mg/mL (~87.31 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (8.73 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 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 is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume 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 (8.73 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 is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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 (8.73 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 is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.4925 mL	17.4624 mL	34.9247 mL
	5 mM	0.6985 mL	3.4925 mL	6.9849 mL
	10 mM	0.3492 mL	1.7462 mL	3.4925 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.