M62812 is a toll-like receptor 4 (TLR4) signaling inhibitor. M62812 inhibits endothelial cell and leukocyte activation and prevents lethal septic shock in mice. M62812 can reduce LPS-induced coagulation and inflammatory responses. M62812 may be utilized in sepsis research.

Physicochemical Properties

Molecular Formula	C13H13CL2N3OS
Molecular Weight	330.23
Exact Mass	329.015
CAS #	613263-00-6
Related CAS #	M62812 free base;613262-61-6
PubChem CID	44224214
Appearance	White to off-white solid powder
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	2
Heavy Atom Count	20
Complexity	291
Defined Atom Stereocenter Count	0
InChi Key	SMTKPLISDFNJDJ-UHFFFAOYSA-N
InChi Code	InChI=1S/C13H11N3OS.2ClH/c14-10-3-1-2-4-11(10)17-8-5-6-9-12(7-8)18-16-13(9)15;;/h1-7H,14H2,(H2,15,16);2*1H
Chemical Name	6-(2-aminophenoxy)-1,2-benzothiazol-3-amine;dihydrochloride
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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.
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	With an IC50 of 2.4 μg/mL, M62812 (10 μg/mL; 6 h) totally blocks LPS-induced NF-κB activation in NF-κB luciferase-expressing cells[1]. With an IC50 of 0.7 μg/mL, M62812 (3 μg/mL; 6 h) totally suppresses TNF-α production in peripheral blood mononuclear cells stimulated by LPS[1]. In human endothelial cells, M62812 (3 μg/mL; 6 h) totally suppresses the production of IL-6 and E-selection, with IC50 values of 0.43 μg/mL and 1.4 μg/mL, respectively [1].
ln Vivo	M62812 (iv; 10–20 mg/kg; single dose) protects against endotoxic shock in a D-galactosamine-sensitized mice model by lowering coagulation and inflammatory markers[1]. By using the mouse cecal ligation and puncture paradigm, M62812 (iv; 20 mg/kg; once daily for three days) avoids mouse death[1].
Animal Protocol	Animal/Disease Models: D-galactosamine-sensitized endotoxin shock mouse model[1] Doses: 10-20 mg/kg Route of Administration: intravenous (iv) administration (iv) Experimental Results: Prevented elevation of TNF-α, IL-6, soluble E-selectin, thrombin/ antithrombin complexes and glutamic pyruvic transaminase activity at 20 mg/kg. Prolonged survival in a d-galactosamine-sensitized endotoxin shock mouse model. Animal/Disease Models: Cecal ligation and puncture mouse model[1] Doses: 20 mg/kg Route of Administration: intravenous (iv) administration ( iv); one time/day for three days Experimental Results: decreased mortality in a murine cecal ligation and puncture model.
References	[1]. Toll-like receptor 4 signal transduction inhibitor, M62812, suppresses endothelial cell and leukocyte activation and prevents lethal septic shock in mice. Eur J Pharmacol. 2007 Aug 27;569(3):237-43.

Solubility Data

Solubility (In Vitro)

0.1 M HCL: ≥ 50 mg/mL (151.41 mM) DMSO: 50 mg/mL (151.41 mM) H2O: < 0.1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.57 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 (7.57 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 (7.57 mM) in Saline (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. 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	3.0282 mL	15.1410 mL	30.2819 mL
	5 mM	0.6056 mL	3.0282 mL	6.0564 mL
	10 mM	0.3028 mL	1.5141 mL	3.0282 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.