title: Biopharmaceutical characterization, metabolism, and brain penetration of the triple reuptake inhibitor amitifadine.
abstract: Amitifadine (EB-1010, formerly DOV 21,947) is a serotonin-preferring triple reuptake inhibitor that is a drug candidate for major depressive disorder. We investigated several relevant biopharmaceutic and drug-like characteristics of amitifadine using in vitro methodology and additionally determined the in vivo brain to plasma ratio of the drug in rats. Amitifadine was highly plasma protein bound with over 99% of drug bound to human plasma proteins. Using Caco-2 cell lines, amitifadine was bidirectionally highly permeable and showed no evidence of active secretion. Amitifadine was metabolized slowly by human hepatocytes and the major metabolite was the lactam EB-10101. In vitro studies using human liver microsomes demonstrated that EB-10101 was formed by monoamine oxidase A (MAO-A) and a NADPHdependent enzyme, possibly a cytochrome P450 (CYP) isoform. Amitifadine was a moderate inhibitor of the human isoforms of the major drug metabolizing enzymes CYP2D6, CYP3A4, CYP2C9, and CYP2C19 (IC50 = 9 - 100 μM), but was a potent inhibitor of human CYP2B6 (IC50 = 1.8 μM). The brain to plasma ratio for amitifadine varied from 3.7 - 6.5 at various time points, indicating preferential partitioning into rat brain versus plasma. The low affinity for the major drug metabolizing CYP enzymes and metabolism by multiple pathways may reduce pharmacokinetic drug-drug interactions and effects of enzyme polymorphisms. Overall, these studies suggest that amitifadine has drug-like characteristics favorable for drug development.
authors.forename: Frank P
authors.initials: FP
authors.lastname: Bymaster
authors.forename: Piyun
authors.initials: P
authors.lastname: Chao
authors.forename: Heidi
authors.initials: H
authors.lastname: Schulze
authors.forename: Pierre V
authors.initials: PV
authors.lastname: Tran
authors.affiliation: Euthymics Bioscience Inc, 43 Thorndike St. Suite 1-3, Cambridge, MA, 02141, USA. fbymaster@euthymics.com.
authors.forename: Randall D
authors.initials: RD
authors.lastname: Marshall
mesh_terms: D000818:Animals; D000928:Antidepressive Agents / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics*; D001189:Aryl Hydrocarbon Hydroxylases; D001372:Aza Compounds / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics*; D001702:Biopharmaceutics; D001798:Blood Proteins / Q000378:metabolism; D001921:Brain / Q000378:metabolism*; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics*; D018938:Caco-2 Cells; D002463:Cell Membrane Permeability; D065731:Cytochrome P-450 CYP2C19; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D038181:FMN Reductase / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D007769:Lactams / Q000378:metabolism; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism; D008995:Monoamine Oxidase / Q000378:metabolism; D009249:NADP / Q000502:physiology; D014179:Neurotransmitter Uptake Inhibitors / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics*; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: C474071:1-(3,4-dichlorophenyl)-3-azabicyclo-(3.1.0)hexane hydrochloride; D000928:Antidepressive Agents; D001372:Aza Compounds; D001798:Blood Proteins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D065607:Cytochrome P-450 Enzyme Inhibitors; D004791:Enzyme Inhibitors; D007769:Lactams; D014179:Neurotransmitter Uptake Inhibitors; D009249:NADP; D003577:Cytochrome P-450 Enzyme System; D001189:Aryl Hydrocarbon Hydroxylases; C045793:CYP2C19 protein, human; D065731:Cytochrome P-450 CYP2C19; D008995:Monoamine Oxidase; D038181:FMN Reductase