PT - JOURNAL ARTICLE AU - Coe, Kevin J. AU - Feinstein, Mark AU - Higgins, J. William AU - Leung, Perry AU - Scott, Brian P AU - Skaptason, Judith AU - Tam, Yuen AU - Volak, Laurie P AU - Kinong, Jennifer AU - Bittner, Anton AU - McAllister, Heather AU - Lim, Nathan M AU - Hack, Michael AU - Koudriakova, Tatiana TI - <strong><strong>Characterization of JNJ-2482272 [4-(4-methyl-2-(4-(trifluoromethyl)phenyl)thiazole-5-yl)pyrimidine-2-amine] as a Strong AhR Activator in Rat and Human</strong></strong> AID - 10.1124/dmd.121.000825 DP - 2022 Jan 01 TA - Drug Metabolism and Disposition PG - DMD-AR-2021-000825 4099 - http://dmd.aspetjournals.org/content/early/2022/06/09/dmd.121.000825.short 4100 - http://dmd.aspetjournals.org/content/early/2022/06/09/dmd.121.000825.full AB - JNJ-2482272, under investigation as an anti-inflammatory agent, was orally administered to rats q.d. at 60 mg/kg for six consecutive days. Despite high plasma exposure after single administration (Cmax of 7.1 µM), JNJ-2482272 had plasma concentration beneath the lower limit of quantification (3 ng/mL) after six consecutive days of dosing. To determine if JNJ-2482272 is an autoinducer in rats, plated rat hepatocytes were treated with JNJ-2482272 for two days. The major hydroxylated metabolites of JNJ-2482272 were isolated and characterized by MS and NMR analyses. Compared to the vehicle treated cells, a concentration-dependent increase was observed in the formation of Phase I and II-mediated metabolites coinciding with greater expression of P450s and UGTs in rat hepatocytes. CYP1A1, CYP1A2, CYP1B1, and UGT1A6 transcripts were predominantly induced, suggesting that JNJ-2482272 is an activator of the aryl hydrocarbon receptor (AhR). In a human AhR reporter assay, JNJ-2482272 demonstrated potent AhR activation with an EC50 value of 0.768 nM, a potency more comparable to the strong AhR activator and toxin 2,3,7,8-tetrachloro-dibenzodioxin (TCDD) than to weaker AhR activators 3-methylcholanthrene (3-MC), β-naphthoflavone (βNF), and omeprazole (OME). In plated human hepatocytes, JNJ-2482272 induced CYP1A1 gene expression with an EC50 of 20.4 nM and increased CYP1A activity &gt; 50-fold from basal levels. In human recombinant P450s (rCYPs), JNJ-2482272 was exclusively metabolized by the CYP1 family of enzymes and most rapidly by CYP1A1. The summation of these in vitro findings bridges the in vivo conclusion that JNJ-2482272 is a strong autoinducer in rats and potentially in humans through potent AhR activation. Significance Statement Drugs that induce their own metabolism (autoinducers) can pose challenges in drug development due to their lack of sustained exposures for pharmacology and safety assessment often precluding their development. JNJ-2482272 is demonstrated herein as a strong AhR activator and CYP1A autoinducer, explaining its near complete loss of exposure after repeat administration in rat, that is likely translatable to human (if progressed further) considering its nanomolar potency comparable to “classical” AhR ligands like TCDD despite bearing a “non-classical” drug structure.