2614W94 

MAO-A and -B forms are assayed. Rat brain mito-chondrial extract is pre-incubated with the inhibitor for 15 min at 37°C in 50 mM potassium phosphate buffer (pH 7.4). Substrates [³H]serotonin (0.2 mM, 5 Ci/mol) and [¹⁴C]β-phene-thylamine (10 µM, 3 Ci/mol) are then added, and incubation at 37°C is continued for 20 min. Blank assays contain 2 mM pargyline to inhibit all MAO activity. The reaction is terminated with 0.2 mL of 2 N HCl, and products are extracted with 6 mL of ethyl acetate/toluene (1:1). A 4 mL aliquot of the organic layer is countedin 10 mL of Ecolite in a scintillation spectrometer programmed for double-label counting. Assays are performed in triplicate unless otherwise indicated. At the above concentrations, serotonin is a selective substrate for MAO-A, and β-phenethylamine is aselective substrate for MAO-B.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Rats: Nonfasted Sprague-Dawley male rats (250-350 g) are dosed by gavage with 0.5% methyl cellulose or with 2614W94 or other compounds suspended in the methyl cellulose vehicle. For all groups, n = 3 unless otherwise specified. For oral administration, dosing volume is 10 mL/kg of body weight. For intravenous dosing, the vehicle is a mixture of PEG 400 (polyethylene glycol; molecular weight, 400), ethanol, and physiologic saline in a volume ratio of 1.5/1.5/1.0, respectively, and the dosing volume is 1 mL/kg. After dosing, rats are returned to their cages and allowed free access to water. Any animals kept overnight are also given food. Death is by CO₂ asphyxiation, after which brains and livers are promptly removed, frozen on dry ice, and stored at -70°C.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Helen L. White, et al. Biochemical and Pharmacologic Properties of 2614W94, a Reversible, Competitive Inhibitor of MonoamineOxidase-A. DRUG DEVELOPMENT RESEARCH 45:1-9 (1998).