title: Identification of the cytochrome P450 and other enzymes involved in the in vitro oxidative metabolism of a novel antidepressant, Lu AA21004.
abstract: 1-[2-(2,4-Dimethyl-phenylsulfanyl)-phenyl]-piperazine (Lu AA21004) is a novel antidepressant that is currently in late-stage clinical development for major depressive disorder. In the present study, the metabolism of Lu AA21004 was investigated using human liver microsomes (HLM), human liver S9 fraction, and recombinant enzymes. Lu AA21004 was found in vitro to be oxidized to a 4-hydroxy-phenyl metabolite, a sulfoxide, an N-hydroxylated piperazine, and a benzylic alcohol, which was further oxidized to the corresponding benzoic acid [3-methyl-4-(2-piperazin-1-yl-phenysulfanyl)-benzoic acid (Lu AA34443)]. The formation of the 4-hydroxy-phenyl metabolite was catalyzed by CYP2D6 with some contribution from CYP2C9, whereas the formation of the sulfoxide was mediated by CYP3A4/5 and CYP2A6. CYP2C9 and CYP2C19 were the primary enzymes responsible for formation of the N-hydroxylated metabolite. The benzylic alcohol was formed by CYP2D6 only. The oxidation of the benzylic alcohol to the corresponding benzoic acid of Lu AA21004 was catalyzed by alcohol dehydrogenase and aldehyde dehydrogenase, with some contribution from aldehyde oxidase. CYP2D6 was also capable of catalyzing the formation of the benzoic acid of Lu AA21004; however, its overall contribution to this pathway was negligible. Enzyme kinetic parameters revealed that the rate-limiting step in the formation of the benzoic acid from Lu AA21004 is the formation of the corresponding alcohol. Thus, the intrinsic clearance (V(max)/K(m)) in HLM for metabolism of Lu AA21004 to the benzylic alcohol was 1.13 × 10(-6) l · min(-1) · mg(-1), whereas the subsequent metabolism of the benzylic alcohol to the benzoic acid of Lu AA21004 is characterized by an intrinsic clearance (V(max)/K(m)) in S9 fraction of 922 × 10(-6) l · min(-1) · mg(-1).
authors.affiliation: Department of Drug Metabolism, H. Lundbeck A/S, Ottiliavej 9, 2500 Valby, Denmark. mgh@lundbeck.com
authors.forename: Mette G
authors.initials: MG
authors.lastname: Hvenegaard
authors.forename: Benny
authors.initials: B
authors.lastname: Bang-Andersen
authors.forename: Henrik
authors.initials: H
authors.lastname: Pedersen
authors.forename: Morten
authors.initials: M
authors.lastname: Jørgensen
authors.forename: Ask
authors.initials: A
authors.lastname: Püschl
authors.forename: Lars
authors.initials: L
authors.lastname: Dalgaard
mesh_terms: D000928:Antidepressive Agents / Q000493:pharmacokinetics* / Q000494:pharmacology*; D002460:Cell Line; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D003865:Major Depressive Disorder / Q000188:drug therapy / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D006900:Hydroxylation; D007527:Isoenzymes; D008862:Microsomes, Liver / Q000201:enzymology* / Q000378:metabolism*; D010084:Oxidation-Reduction; D010879:Piperazines / Q000493:pharmacokinetics* / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000378:metabolism; D013440:Sulfides / Q000493:pharmacokinetics* / Q000494:pharmacology*; D000078784:Vortioxetine
chemical_list: D000928:Antidepressive Agents; D007527:Isoenzymes; D010879:Piperazines; D011994:Recombinant Proteins; D013440:Sulfides; D000078784:Vortioxetine; D003577:Cytochrome P-450 Enzyme System