All animal protocols were approved by the University of California Davis Animal Use and Care Committee, and experiments were carried out in compliance with the National Institutes of Health Guide for the use and care of laboratory animals. Upon arrival to the vivarium, 7 week old mice (Swiss Webster mice, Charles River Laboratories, Wilmington, MA) were acclimated to their new housing environment for one week and kept under a 12 hour light/dark cycle with free access to food and water.
DZP (5 mg/mL, Hospira, Inc., Lake Forest, IL) was dissolved in 40% propylene glycol, 10 % ethanol and saline. TPPU (1-trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl) urea) was synthesized [18 (link)], purified and characterized in-house (a kind gift of Dr Sing Lee from UC Davis) and was dissolved in PEG400 to obtain a clear solution. Mice were injected (i.p.) either with DZP alone at 1, 3 or 10 mg/kg or in combination with TPPU at 3 mg/kg i.p. one hour before administration of DZP at the doses indicated. Midazolam (Hospira, Inc., Lake Forest, IL) was administered at 1.8 and 10 mg/kg doses by intramuscular route using saline as vehicle. Animals that served as controls were given the corresponding vehicle solutions, PEG400 for TPPU or 40% propylene glycol, 10% ethanol in saline for DZP and saline for Midazolam injections.
DZP (5 mg/mL, Hospira, Inc., Lake Forest, IL) was dissolved in 40% propylene glycol, 10 % ethanol and saline. TPPU (1-trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl) urea) was synthesized [18 (link)], purified and characterized in-house (a kind gift of Dr Sing Lee from UC Davis) and was dissolved in PEG400 to obtain a clear solution. Mice were injected (i.p.) either with DZP alone at 1, 3 or 10 mg/kg or in combination with TPPU at 3 mg/kg i.p. one hour before administration of DZP at the doses indicated. Midazolam (Hospira, Inc., Lake Forest, IL) was administered at 1.8 and 10 mg/kg doses by intramuscular route using saline as vehicle. Animals that served as controls were given the corresponding vehicle solutions, PEG400 for TPPU or 40% propylene glycol, 10% ethanol in saline for DZP and saline for Midazolam injections.
