title: The peroxisome proliferator-activated receptor α agonist, AZD4619, induces alanine aminotransferase-1 gene and protein expression in human, but not in rat hepatocytes: Correlation with serum ALT levels.
abstract: Alanine aminotransferase (ALT) in serum is the standard biomarker for liver injury. We have previously described a clinical trial with a novel selective peroxisome proliferator-activated receptor α (PPARα) agonist (AZD4619), which unexpectedly caused increased serum levels of ALT in treated individuals without any other evidence of liver injury. We pinpointed a plausible mechanism through which AZD4619 could increase serum ALT levels; namely through the PPARα-specific activation of the human ALT1 gene at the transcriptional level. In the present study, we present data from the preceding rat toxicity study, demonstrating that AZD4619 had no effect on rat serum ALT activity levels, and further experiments were performed to elucidate the mechanisms responsible for this species-related difference. Our results revealed that AZD4619 increased ALT1 protein expression in a dose-dependent manner in human, but not in rat primary hepatocytes. Cloning of the human and rat ALT1 promoters into luciferase vectors confirmed that AZD4619 induced only the human, but not the rat ALT1 gene promoter in a dose-dependent manner. In PPARα-GAL4 reporter gene assays, AZD4619 was >100-fold more potent on the human vs. rat PPARα levels, explaining the differences in induction of the ALT1 gene between the species at the concentration range tested. These data demonstrate the usefulness of the human and rat ALT1 reporter gene assays for testing future drug candidates at the preclinical stage. In drug discovery projects, these assays elucidate whether elevations in ALT levels observed in vivo or in the clinic are due to metabolic effects rather than a toxic event in the liver.
authors.affiliation: Drug Safety and Metabolism, AstraZeneca, 43183 Mölndal, Sweden.
authors.forename: Petra
authors.initials: P
authors.lastname: Thulin
authors.affiliation: Bioscience, AstraZeneca, 43183 Mölndal, Sweden.
authors.forename: Krister
authors.initials: K
authors.lastname: Bamberg
authors.affiliation: Drug Safety and Metabolism, AstraZeneca, 43183 Mölndal, Sweden.
authors.forename: Marcin
authors.initials: M
authors.lastname: Buler
authors.affiliation: Drug Safety and Metabolism, AstraZeneca, 43183 Mölndal, Sweden.
authors.forename: Björn
authors.initials: B
authors.lastname: Dahl
authors.affiliation: Drug Safety and Metabolism, AstraZeneca, 43183 Mölndal, Sweden.
authors.forename: Björn
authors.initials: B
authors.lastname: Glinghammar
mesh_terms: D000410:Alanine Transaminase / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008297:Male; D047493:PPAR alpha / Q000819:agonists*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011422:Propionates / Q000494:pharmacology*; D017208:Rats, Wistar; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D013450:Sulfones / Q000494:pharmacology*; D015262:Xenobiotics / Q000494:pharmacology*
chemical_list: C000615714:2-(2-(4-methylphenyl)ethylsulfanyl)-3-(4-(2-(4-methylsulfonyloxyphenoxy)ethyl)phenyl)propanoic acid; D047493:PPAR alpha; D011422:Propionates; D013450:Sulfones; D015262:Xenobiotics; D008156:Luciferases; D000410:Alanine Transaminase