PNU-120596

2 years -80 in solvent

PNU-120596 is intravenously administrated 5 minutes before auditory gating measurements.

SH-SY5Y-alpha7 cells

1 mg/kg

216 nM (EC50) [1], 216 nM (EC50) [1], 216 nM (EC50) [1], 216 nM (EC50) [1], 216 nM (EC50) [1], 216 nM (EC50) [1]

Systemic administration of PNU-120596 (1 mg/kg) to rats improves the auditory gating deficit caused by amphetamine, a model proposed to reflect a circuit level disturbance associated with schizophrenia. [1] When administered prior to Carrageenan, 30 mg/kg PNU-1230596 significantly blunts mechanical hyperalgesia and weight bearing deficits for up to 4 hours. PNU-120596 attenuates the carrageenan-induced increase in levels of TNF-alpha and IL-6 within the hindpaw oedema, diclofenac only attenuated IL-6 levels. Established mechanical hyperalgesia induced by Carrageenan or CFA is also partially reversed by PNU-120596. [4]

Ca2+ Fluorescence Assay, SH-EP1 human epithelial cells expressing a variant of thealpha7 nAChR (alpha7*) are grown in minimal essential medium (MEM) containing nonessential amino acids supplemented with 10% fetal bovine serum, L-glutamine, 100 U/ml penicillin/streptomycin, 250 ng/mL fungizone, 400 ug/mL hygromycin B, and 800 ug/mL geneticin. alpha7* is a variant of the human alpha7 nAChR, with two point mutations in the first transmembrane domain (T230P and C241S) that allow for high functional expression in SH-EP1 cells [Groppi VE, Wolfe ML, Berkenpas MB (2003) U.S. Patent 6, 693, 172 B1]. Cells are grown in a 37 C incubator with 6% CO2. Cells are trypsinized and plated in 96-well plates with dark side walls and clear bottoms at a density of 2, 104 cells/well 2 days before analysis. Cells are loaded with a mixture of Calcium reen-1AM in anhydrous dimethylsulfoxide and 20% pluronic F-127. This reagent is added directly to the growth medium of each well to achieve a final concentration of 2 uM Calcium Green-1 AM. Cells are then incubated in the dye for 1 hour at 37 C and then washed four times with Mark's modified Earle's balanced salt solution (MMEBSS) composed of the following (inmM): 4 CaCl2, 0.8 MgSO4, 20 NaCl, 5.3 KCl, 5.6 D-glucose, 20 Tris-HEPES, and 120 N-methyl-D-glucamine, pH 7.4. After the fourth cycle, the cells are allowed to incubate at 37 C for at least 10 minutes. The final volume of MMEBSS in each well is 100 uL and atropine is added to all wells for a final concentration of 1 uM. A fluorometric imaging plate reader (FLIPR, Molecular Devices, Union City, CA) is set up to excite Calcium Green at 488 nm using 500 mW of power and reading fluorescence emission of >525 nm. A 0.5 seconds exposure is used to illuminate each well. Fluorescence is detected using an F-stop set of either 2.0 or 1.2. After 30 seconds of baseline recording, test compounds are added to each well of a 96-well plate in 50 uL of a 3, stock. In each experiment, four wells are used as vehicle (0.2% DMSO) controls.

DMSO 62 mg/mL, Water <1 mg/mL, Ethanol 1 mg/mL

1-(5-chloro-2, 4-dimethoxyphenyl)-3-(5-methylisoxazol-3-yl)urea