title: In vitro cytotoxicity assay to evaluate the toxicity of an electrophilic reactive metabolite using glutathione-depleted rat primary cultured hepatocytes.
abstract: Glutathione plays an important role as not only a scavenger of reactive oxygen species but also in the conjugation or detoxification of electrophilic reactive metabolites, which has been thought to be one of the causes for idiosyncratic drug toxicity (IDT). Therefore, toxic responses to the reactive metabolites have been expected to be expressed more strongly in a glutathione-depleted condition. In the present study, we attempted to establish an in vitro cytotoxicity assay method to evaluate the toxicity of the reactive metabolite using rat primary cultured hepatocytes with cellular glutathione depletion by l-buthionine-S,R-sulfoximine. Also, we investigated whether the IDT risk is predictable by comparing the cytotoxic sensitivity between glutathione-depleted hepatocytes and untreated hepatocytes. Consequently, 10 drugs of 42 approved drugs, which were classified into 4 IDT categories (Withdrawn, Black box warning, Warning, and Safe), demonstrated higher cytotoxic sensitivity in the glutathione-depleted hepatocytes. Furthermore, a correlation was observed between the incidence of drugs with higher cytotoxic sensitivity in the glutathione-depleted hepatocytes and the IDT risk. The incidence was 50% in the Withdrawn category, 38% in the Black box warning category, 22% in the Warning category, and 8% in the Safe category. These results suggest that the IDT risk of some drugs may be predicted by comparing the cytotoxic sensitivity between them. Additionally, this method may be useful as a screening in the early stage of drug development where leads/candidates are optimized.
authors.affiliation: Daiichi-Sankyo Co., Ltd., Fukuroi, Shizuoka 437-0065, Japan. fujimoto.kazunori.wr@daiichisankyo.co.jp
authors.forename: Kazunori
authors.initials: K
authors.lastname: Fujimoto
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kishino
authors.forename: Takashi
authors.initials: T
authors.lastname: Yamoto
authors.forename: Sunao
authors.initials: S
authors.lastname: Manabe
authors.forename: Atsushi
authors.initials: A
authors.lastname: Sanbuissho
mesh_terms: D000082:Acetaminophen / Q000378:metabolism / Q000633:toxicity; D000818:Animals; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002478:Cells, Cultured; D017277:Drug Approval; D064420:Drug-Related Side Effects and Adverse Reactions / Q000378:metabolism*; D005978:Glutathione / Q000172:deficiency*; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D012306:Risk; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D000082:Acetaminophen; D019328:Buthionine Sulfoximine; D005978:Glutathione