title: Inhibition of hepatobiliary transport as a predictive method for clinical hepatotoxicity of nefazodone.
abstract: Treatment with the antidepressant nefazodone has been associated with clinical idiosyncratic hepatotoxicty. Using membranes expressing human bile salt export pump (BSEP), human sandwich hepatocytes, and intact rats, we compared nefazodone and its marketed analogs, buspirone and trazodone. We found that nefazodone caused a strong inhibition of BSEP (IC(50) = 9 microM), inhibition of taurocholate efflux in human hepatocytes (IC(50) = 14 microM), and a transient increase in rat serum bile acids 1 h after oral drug administration. Buspirone or trazodone had no effect on biliary transport system. Nefazodone produced time- and concentration-dependent toxicity in human hepatocytes with IC(50) = 18 microM and 30 microM measured by inhibition of protein synthesis after 6 h and 24 h incubation, respectively. Toxicity was correlated with the amount of unmetabolized nefazodone. Partial recovery in toxicity by 24 h has been associated with metabolism of nefazodone to sulfate and glucuronide conjugates. The saturation of nefazodone metabolism resulted in sustained decrease in protein synthesis and cell death at 50 microM. The toxicity was not observed with buspirone or trazodone. Addition of 1-aminobenzotriazole (ABT), an inhibitor of CYP450, resulted in enhancement of nefazodone toxicity at 10 microM and was associated with accumulation of unmetabolized nefazodone. In human liver microsomes, ABT also prevented metabolism of nefazodone and formation of glutathione conjugates. We suggest that inhibition of bile acid transport by nefazodone is an indicator of potential hepatotoxicity. Our findings are consistent with the clinical experience and suggest that described methodology can be applied in the selection of nonhepatotoxic drug candidates.
authors.affiliation: Department of Safety Science and Pharmacokinetics, Pfizer Global Research and Development, Ann Arbor, Michigan 48105, USA. vsevolod.kostrubsky@pfizer.com
authors.forename: Seva E
authors.initials: SE
authors.lastname: Kostrubsky
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Strom
authors.forename: Amit S
authors.initials: AS
authors.lastname: Kalgutkar
authors.forename: Shaila
authors.initials: S
authors.lastname: Kulkarni
authors.forename: James
authors.initials: J
authors.lastname: Atherton
authors.forename: Rouchelle
authors.initials: R
authors.lastname: Mireles
authors.forename: Bo
authors.initials: B
authors.lastname: Feng
authors.forename: Raylene
authors.initials: R
authors.lastname: Kubik
authors.forename: Janean
authors.initials: J
authors.lastname: Hanson
authors.forename: Ellen
authors.initials: E
authors.lastname: Urda
authors.forename: Abdul E
authors.initials: AE
authors.lastname: Mutlib
mesh_terms: D000818:Animals; D018687:Antidepressive Agents, Second-Generation / Q000633:toxicity*; D001647:Bile Acids and Salts / Q000097:blood / Q000378:metabolism*; D001648:Bile Canaliculi / Q000187:drug effects / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects*; D002065:Buspirone / Q000633:toxicity; D002478:Cells, Cultured; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D010879:Piperazines; D051381:Rats; D017207:Rats, Sprague-Dawley; D014196:Trazodone / Q000633:toxicity; D014230:Triazoles / Q000633:toxicity*
chemical_list: D018687:Antidepressive Agents, Second-Generation; D001647:Bile Acids and Salts; D010879:Piperazines; D014230:Triazoles; C051752:nefazodone; D002065:Buspirone; D014196:Trazodone