GW791343 tri HCl is a potent NAM (negative allosteric modulator) (species-specific) of human P2X7 receptor, which produces noncompetitive antagonism to human P2X7 receptor, with pIC50 of 6.9-7.2. GW791343 tri HCl enhances circadian ATP release. GW791343 tri HCl may be utilized in the research of neurological diseases.

Physicochemical Properties

Molecular Formula	C20H27CL3F2N4O
Molecular Weight	483.810388803482
Exact Mass	482.122
CAS #	309712-55-8
Related CAS #	GW791343 dihydrochloride;1019779-04-4
PubChem CID	9848159
Appearance	Typically exists as solid at room temperature
LogP	6.124
Hydrogen Bond Donor Count	6
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	6
Heavy Atom Count	30
Complexity	475
Defined Atom Stereocenter Count	0
SMILES	C(NC1=CC(CN2CCNCC2)=CC=C1C)(=O)CNC1=CC=C(F)C(F)=C1.[H]Cl.[H]Cl.[H]Cl
InChi Key	WSBRAHWNJBXXJM-UHFFFAOYSA-N
InChi Code	InChI=1S/C20H24F2N4O.3ClH/c1-14-2-3-15(13-26-8-6-23-7-9-26)10-19(14)25-20(27)12-24-16-4-5-17(21)18(22)11-16;;;/h2-5,10-11,23-24H,6-9,12-13H2,1H3,(H,25,27);3*1H
Chemical Name	2-(3,4-difluoroanilino)-N-[2-methyl-5-(piperazin-1-ylmethyl)phenyl]acetamide;trihydrochloride
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	P2X7 Receptor 6.9-7.2 (pIC50)
ln Vitro	GW791343 trihydrochloride (0.01, 0.03, 0.1, 0.3, 1, 3, 10 µM; 40 min) has antagonistic action that is non-competitive towards the human P2X7 receptor[1]. The human P2X7 receptor exhibits an anegative allosteric modulator activity in response to GW791343 trihydrochloride (3, 10, 30 µM; 40 min)[1]. The ATP rhythm in SCN cells is improved by GW791343 trihydrochloride (5 µM; 24-48 h; ATP measured every 4 h)[2].
Cell Assay	Cell Viability Assay[1] Cell Types: HEK293 cells (expressing human recombinant P2X7 receptors). Tested Concentrations: 0.01, 0.03, 0.1, 0.3, 1, 3, 10 µM. Incubation Duration: 40 min (pre-incubate for 10 min and incubate with other P2X7 receptor antagonists for another 30 min). Experimental Results: Inhibited agonist-stimulated ethidium accumulation in both sucrose and NaCl buffer. diminished maximal responses toATP and BzATP in sucrose buffer. Cell Viability Assay[1] Cell Types: HEK293 cells (expressing human recombinant P2X7 receptors). Tested Concentrations: 3 , 10, 30 µM. Incubation Duration: 40 min (pre-incubate for 10 min and incubate with other P2X7 receptor antagonists for another 30 min). Experimental Results: demonstrated slow reversal effects at the human P2X7 receptor (after 45 min had reversed sufficiently) , and had a rapid dissociation rate. Cell Viability Assay[2] Cell Types: SCN cells (from 16-to 21- day-old Wistar rats, which are kept under a controlled 12-12 h light-dark cycle from birth). Tested Concentrations: 5 µM (replace the medium
References	[1]. Negative and positive allosteric modulators of the P2X(7) receptor. Br J Pharmacol. 2008 Feb;153(4):737-50. [2]. Circadian ATP Release in Organotypic Cultures of the Rat Suprachiasmatic Nucleus Is Dependent on P2X7 and P2Y Receptors. Front Pharmacol. 2018 Mar 6;9:192.

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	2.0669 mL	10.3346 mL	20.6693 mL
	5 mM	0.4134 mL	2.0669 mL	4.1339 mL
	10 mM	0.2067 mL	1.0335 mL	2.0669 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.