Benztropine mesylate

Jump-In T-REx HEK 293 cells modified for doxycycline-inducible overexpression of the wild-type human norepinephrine transporter (JumpIn-NET) were provided by CeMM (Research Center for Molecular Medicine, Medical University of Vienna, Austria). These cells were used in the TRACT and the fluorescent substrate uptake assays to allow a good comparison. Benztropine mesylate, desipramine hydrochloride, Dulbecco’s Modified Eagle’s Medium (DMEM), dopamine hydrochloride, doxycycline hyclate, (−)-epinephrine (+)-bitartrate salt, L-(−)-norepinephrine (+)-bitartrate salt monohydrate and (±)-propranolol hydrochloride were purchased from Sigma Aldrich (St. Louis, MO, USA). Nisoxetine hydrochloride, maprotiline hydrochloride and yohimbine hydrochloride were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). GBR12909 dihydrochloride and reboxetine mesylate were purchased from Toronto Research Chemicals (Toronto, ON, Canada). Amitriptyline hydrochloride, atomoxetine hydrochloride, bupropion hydrochloride, milnacipran hydrochloride and nortriptyline hydrochloride were purchased from Tebu-Bio (Heerhugowaard, The Netherlands). Cocaine hydrochloride was purchased from Duchefa Farma (Haarlem, The Netherlands), where Leiden University has been certified for its use in pharmacological experiments. All other chemicals were of analytical grade and obtained from standard commercial sources.

Drugs: Methiopropamine hydrochloride (HCl) (mol. wt. 191.7) was purchased from Cayman Chemicals (Middlesex, UK) and (+)methamphetamine HCl (mol. wt. 185.7) was purchased from Chiron AS (Trondheim, Norway). Chemicals: 5-HT HCl, L-NE HCl, 5-HIAA HCl, 3-MT HCl, ephedrine-D₃, sodium fluoride, methanol (LC-MS Chromasolv^®), ammonium formate, benztropine mesylate, reboxetine mesylate hydrate, fluoxetine HCl and adenosine 5′-triphosphate disodium salt hydrate were purchased from Sigma-Aldrich (Oslo, Norway). DA HCl and DOPAC were purchased from Fluka (Buchs, Switzerland). DA-¹³C₆ HCl, 5-HT-D₄ HCl, 5-HIAA-D₅, methamphetamine-¹³C₆ and nor-methiopropamine were purchased from Chiron AS. Di-sodiumtetraborate-10-hydrate and ethyl acetate were purchased from Chemi-Teknik AS (Oslo, Norway). Acetic acid and formic acid were purchased from VWR International AS (Oslo, Norway). Sodium hydroxide and n-heptane were purchased from Merck Millipore (Oslo, Norway). Dihydroxyphenylethylamine 3,4-[ring-2,5,6-³H]-(DA) ([³H]DA), NE HCl DL-[7-³H(N)] ([³H]NE), 5-HT [³H(G)] ([³H]5-HT) and UltimaGold^TM liquid scintillation cocktail were purchased form from PerkinElmer Inc. (Oslo, Norway). HEPES was purchased from Thermo Fischer Scientific (Oslo, Norway) and sodium heparin was purchased from Leo Pharma (Oslo, Norway).