The anti-pruritic activity of JTE-907 was studied in NC mice: 1 and 10 mg/kg/day for 20 days

CHO cells expressing CB receptors were harvested and cultured at a density of 1 3 10^4 cells/well in 96-well culture plate. After 24-h culture at 37°C, cells were washed with phosphatebuffered saline and incubated at 37°C for 10 min in HEPES buffer [137 mM NaCl, 4.5 mM KCl, 1.2 mM MgCl2, 1 mM CaCl2 x 2H2O, 20 mM HEPES, and 10 mM D-(+)-glucose at pH 7.4] containing 0.25 mM Ro20-1724 in the presence or absence of JTE-907 (0.1nM-100mkM). Cells were then incubated with 5 mM forskolin at 37°C for 15 min, followed by the addition of ice-cold 2.5% dodecyltrimethylammonium bromide (Amersham Pharmacia Biotech, Piscataway, NJ) to stop the reaction. The wells were agitated for 60 min at room temperature. cAMP concentration in the medium of each well was measured by enzyme immunoassay kit