Cot inhibitor-2 is a potent, selective, orally bioactive cot (Tpl2/MAP3K8) inhibitor (antagonist) with IC50 of 1.6 nM. Cot inhibitor-2 inhibits LPS-stimulated TNF-α production in human whole blood with IC50 of 0.3 μM.

Physicochemical Properties

Molecular Formula	C26H25N8FCL2
Molecular Weight	539.4347
Exact Mass	538.156
CAS #	915363-56-3
PubChem CID	44143370
Appearance	Light yellow to green yellow solid powder
Density	1.45
Boiling Point	713.3ºC at 760 mmHg
Flash Point	385.2ºC
Index of Refraction	1.702
LogP	6.24
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	7
Heavy Atom Count	37
Complexity	792
Defined Atom Stereocenter Count	0
InChi Key	PHNZIIMWDVXPGG-UHFFFAOYSA-N
InChi Code	InChI=1S/C26H25Cl2FN8/c1-2-36-7-5-20(6-8-36)37-15-19(34-35-37)14-31-18-9-21-25(33-17-3-4-24(29)22(27)10-17)16(12-30)13-32-26(21)23(28)11-18/h3-4,9-11,13,15,20,31H,2,5-8,14H2,1H3,(H,32,33)
Chemical Name	8-chloro-4-(3-chloro-4-fluoroanilino)-6-[[1-(1-ethylpiperidin-4-yl)triazol-4-yl]methylamino]quinoline-3-carbonitrile
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 Vivo	At a dosage of 100 mg/kg, Cot Inhibitor-2 (Compound 34) had a Cmax of 517 ng/mL (0.89 μM) and an AUC of 4841 ng h/mL. In a paradigm where LPS causes TNF-α production in female Sprague-Dawley rats, Cot inhibitor-2 was evaluated. Cot inhibitor-2 inhibits LPS-induced TNF-α by 83% at an injury dose of 25 mg/kg [1].
References	[1]. Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood. Bioorg Med Chem Lett. 2009 Jul 1;19(13):3485-8.
Additional Infomation	8-chloro-4-(3-chloro-4-fluoroanilino)-6-[[1-(1-ethyl-4-piperidinyl)-4-triazolyl]methylamino]-3-quinolinecarbonitrile is an aminoquinoline.

Solubility Data

Solubility (In Vitro)

DMSO : ~40 mg/mL (~74.15 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 3 mg/mL (5.56 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 30.0 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: ≥ 3 mg/mL (5.56 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 30.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 3: ≥ 2 mg/mL (3.71 mM) (saturation unknown) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.8538 mL	9.2690 mL	18.5381 mL
	5 mM	0.3708 mL	1.8538 mL	3.7076 mL
	10 mM	0.1854 mL	0.9269 mL	1.8538 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.