ASK1-IN-2 is a potent, orally bioactive inhibitor of apoptosis signal-regulated kinase 1 (ASK1) with IC50 of 32.8 nM. ASK1-IN-2 may be used in ulcerative colitis research.

Physicochemical Properties

Molecular Formula	C19H17FN6O
Molecular Weight	364.3763
Exact Mass	364.144
CAS #	2541792-70-3
PubChem CID	155671000
Appearance	White to off-white solid powder
LogP	2.5
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	4
Heavy Atom Count	27
Complexity	534
Defined Atom Stereocenter Count	0
SMILES	FC1C([H])=C([H])C2C([H])=C(C(N([H])C3=C([H])C([H])=C([H])C(C4=NN=C([H])N4C([H])(C([H])([H])[H])C([H])([H])[H])=N3)=O)N([H])C=2C=1[H]
InChi Key	LYUQNMNUBOUCBX-UHFFFAOYSA-N
InChi Code	InChI=1S/C19H17FN6O/c1-11(2)26-10-21-25-18(26)14-4-3-5-17(23-14)24-19(27)16-8-12-6-7-13(20)9-15(12)22-16/h3-11,22H,1-2H3,(H,23,24,27)
Chemical Name	6-fluoro-N-[6-(4-propan-2-yl-1,2,4-triazol-3-yl)pyridin-2-yl]-1H-indole-2-carboxamide
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	ASK1 32.8 nM (IC50)
ln Vitro	In AP1-HEK293 cells, ASK1-IN-2 (compound 19) (10 mM; 1 h) reduces luciferase reporter activity with a 95.59% suppression rate[1].
ln Vivo	In mice, dextran sulphate sodium (DSS)-induced ulcerative colitis (UC) is ameliorated by ASK1-IN-2 (25 mg/kg; po daily for 7 d)[1]. In mouse colon tissues treated with DSS, ASK1-IN-2 (25 mg/kg; po daily for 7 days) inhibits the ASK1-p38/JNK signaling pathways and lowers inflammatory cytokine levels[1]. In rats, ASK1-IN-2 (1 mg/kg; iv) has a moderate half-life (T1/2=1.45 h) and poor clearance (CL=1.38 L/h/kg[1]. Rats treated with ASK1-IN-2 (10 mg/kg; po) exhibit high oral exposure (AUClast=4517 h·ng/mL), 62.2% oral bioavailability, and a tolerable terminal half-life (T1/2=2.31 h)[1].
Animal Protocol	Animal/Disease Models: Male ICR mice (18 -22 g, 6-8 weeks) were given 3% DSS (w/v) orally in drinking water[1] Doses: 25 mg/kg Route of Administration: Po daily for 7 days Experimental Results: Induced a significant recovery of body weight loss, with an increase of +11.2%. diminished the disease activity index (DAI) score about a 2 unit. Dramatically prevented colon shortening. Attenuated a severe colonic tissue damage and infiltration of inflammatory cells. Animal/Disease Models: Male SD rats [1] Doses: 1 mg/kg for iv; 10 mg/kg for po (pharmacokinetic/PK Analysis) Route of Administration: Iv and po administration Experimental Results: Iv: CL=1.38 L/h/kg; T1/2=1.45 h. Po: AUClast=4517 h• ng/mL; F=62.2%; T1/2=2.31 h.
References	[1]. Structure-based discovery of 1H-indole-2-carboxamide derivatives as potent ASK1 inhibitors for potential treatment of ulcerative colitis. Eur J Med Chem. 2020 Dec 24;211:113114.

Solubility Data

Solubility (In Vitro)

DMSO : 250 mg/mL (686.10 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (5.71 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 20.8 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: ≥ 2.08 mg/mL (5.71 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 20.8 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	2.7444 mL	13.7219 mL	27.4439 mL
	5 mM	0.5489 mL	2.7444 mL	5.4888 mL
	10 mM	0.2744 mL	1.3722 mL	2.7444 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.