title: In vitro evaluation of hepatotoxic drugs in human hepatocytes from multiple donors: Identification of P450 activity as a potential risk factor for drug-induced liver injuries.
abstract: A possible risk factor for drug-induced hepatotoxicity is drug metabolizing enzyme activity, which is known to vary among individuals due to genetic (genetic polymorphism) and environmental factors (environmental pollutants, foods, and medications that are inhibitors or inducers of drug metabolizing enzymes). We hypothesize that hepatic cytochrome P450-dependent monooxygenase (CYP) activity is one of the key risk factors for drug induced liver injuries (DILI) in the human population, especially for drugs that are metabolically activated to cytotoxic/reactive metabolites. Human hepatocytes from 19 donors were evaluated for the activities of 8 major P450 isoforms: CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1 and CYP3A4. Extensive individual variations were observed, consistent with what is known to be in the human population. As CYP3A4 is known to be one of the most important P450 isoforms for drug metabolism, studies were performed to evaluate the relationship between the in vitro cytotoxicity of hepatotoxic drugs and CYP3A4 activity. In a proof of concept study, hepatocytes from six donors (lots) representing the observed range of CYP3A4 activities were chosen for the evaluation of in vitro hepatotoxicity of four drugs known to be associated with acute liver failure: acetaminophen, cyclophosphamide, ketoconazole, and tamoxifen. The hepatocytes were cultured in collagen-coated plates and treated with the hepatotoxicants for approximately 24 h, followed by viability determination based on cellular adenosine triphosphate (ATP) contents. HH1023, the lot of hepatocytes with the highest CYP3A4 activity, was found to be the most sensitive to the cytotoxicity of all 4 hepatotoxic drugs, thereby suggesting that high CYP3A4 activity may be a risk factor. To further validate the relationship, a second study was performed with hepatocytes from 16 donors. In this study, the hepatocytes were quantified for CYP3A4 activity at the time of treatment. Results of the second study show confirm the correlation between with high CYP3A4 activity and sensitivity to hepatotoxic drugs. Our results with primary cultured hepatocytes from multiple donors support the hypothesis that elevated P450 activity may be a risk factor for drug-induced liver injuries.
authors.affiliation: In Vitro ADMET Laboratories LLC, 9221 Rumsey Road, Columbia, MD 21045, USA.
authors.forename: Doshi
authors.initials: D
authors.lastname: Utkarsh
authors.affiliation: In Vitro ADMET Laboratories LLC, 9221 Rumsey Road, Columbia, MD 21045, USA.
authors.forename: Carol
authors.initials: C
authors.lastname: Loretz
authors.affiliation: In Vitro ADMET Laboratories LLC, 9221 Rumsey Road, Columbia, MD 21045, USA. Electronic address: lialbert@invitroadmet.com.
authors.forename: Albert P
authors.initials: AP
authors.lastname: Li
mesh_terms: D000082:Acetaminophen / Q000633:toxicity; D000293:Adolescent; D000328:Adult; D000368:Aged; D018712:Analgesics, Non-Narcotic / Q000633:toxicity; D000935:Antifungal Agents / Q000633:toxicity; D002478:Cells, Cultured; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D002648:Child; D003520:Cyclophosphamide / Q000633:toxicity; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004965:Estrogen Antagonists / Q000633:toxicity; D005260:Female; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007166:Immunosuppressive Agents / Q000633:toxicity; D007654:Ketoconazole / Q000633:toxicity; D008099:Liver / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008297:Male; D008875:Middle Aged; D020033:Protein Isoforms / Q000378:metabolism; D013629:Tamoxifen / Q000633:toxicity; D055815:Young Adult
chemical_list: D018712:Analgesics, Non-Narcotic; D000935:Antifungal Agents; D004965:Estrogen Antagonists; D007166:Immunosuppressive Agents; D020033:Protein Isoforms; D013629:Tamoxifen; D000082:Acetaminophen; D003520:Cyclophosphamide; D003577:Cytochrome P-450 Enzyme System; D051544:Cytochrome P-450 CYP3A; D007654:Ketoconazole
keywords: Acetaminophen; CYP3A4; Cyclophosphamide; DILI; Hepatotoxicity; Human hepatocytes; Idiosyncratic hepatotoxicity; Individual differences; Ketoconazole; P450; Risk factors; Tamoxifen