The JAK1/2 Inhibitor Ruxolitinib Reverses Interleukin-6-Mediated Suppression of Drug-Detoxifying Proteins in Cultured Human Hepatocytes

Abstract

The inflammatory cytokine interleukin (IL)-6, which basically activates the Janus kinase (JAK)/ signal transducer and activator of transcription (STAT) signaling pathway, is well known to repress expression of hepatic cytochromes P-450 (P450s) and transporters. Therapeutic proteins, like monoclonal antibodies targeting IL-6 or its receptor, have consequently been demonstrated to restore full hepatic detoxification capacity, which results in inflammatory disease-related drug-drug interactions (idDDIs). In the present study, we investigated whether ruxolitinib, a small drug acting as a JAK1/2 inhibitor and currently used in the treatment of myeloproliferative neoplasms, may also counteract the repressing effects of IL-6 toward hepatic detoxifying systems. Ruxolitinib was found to fully inhibit IL-6-mediated repression of P450 (CYP1A2, CYP2B6, and CYP3A4) and transporter (NTCP, OATP1B1, and OCT1) mRNA levels in primary human hepatocytes and differentiated hepatoma HepaRG cells. Such effects were dose-dependent, with ruxolitinib EC values around 1.0-1.2 M and thus close to ruxolitinib plasma levels that can be reached in patients. Moreover, they were associated with concomitant restoration of P450 and drug transporter activities in IL-6-exposed HepaRG cells. By contrast, ruxolitinib failed to suppress the repression of drug-detoxifying protein mRNA levels caused by IL-1 The JAK inhibitor and anti-rheumatoid arthritis compound tofacitinib was additionally found to reverse IL-6-mediated suppression of P450 and transporter mRNA expressions. Taken together, our results demonstrated that small drugs acting as JAK inhibitors, like ruxolitinib, counteract IL-6-mediated repression of drug-metabolizing enzymes and drug transporters in cultured human hepatocytes. These JAK inhibitors may consequently be hypothesized to restore hepatic detoxification capacity for patients suffering from inflammatory diseases, which may in turn cause idDDIs.

Role of OATP1B1 and OATP1B3 in Drug-Drug Interactions Mediated by Tyrosine Kinase Inhibitors

“…However, such strategies are rarely implemented and reported studies often fail to include positive and negative control inhibitors into the experimental design, which is recommended in the regulatory guidance documents. These issues complicate the interpretation of data and can result in discrepant views on extrapolating from in vitro studies to the clinical situation, as reported for ruxolitinib or crizotinib, where experimental data would suggest statistically significant but not clinically relevant degrees of inhibition [ 67 , 68 , 69 ]. Substrate selection: Since substrate-dependent inhibition by xenobiotics, including TKIs, has been well documented and is acknowledged expressly in the FDA guidance document, the degree to which findings obtained with one particular substrate can be extrapolated to other conditions is uncertain, and potentially accounts for several reported inconsistencies.…”

“…However, such strategies are rarely implemented and reported studies often fail to include positive and negative control inhibitors into the experimental design, which is recommended in the regulatory guidance documents. These issues complicate the interpretation of data and can result in discrepant views on extrapolating from in vitro studies to the clinical situation, as reported for ruxolitinib or crizotinib, where experimental data would suggest statistically significant but not clinically relevant degrees of inhibition [ 67 , 68 , 69 ].…”

Interleukin-6 contributes to the increase in fibroblast growth factor 23 expression in acute and chronic kidney disease

“…UMR106 cells were plated in 96-well tissue culture plates (5 Â 10 4 cells/well) and incubated with HIL-6 (100 ng/ml) 37 or vehicle (0.1% bovine serum albumin). Ruxolitinib (5 mmol/l in ethanol) (Cayman Chemical Company, Ann Arbor, MI) 52 was added without or with HIL-6. HeLa cells in 96-well plates at 80% to 90% confluency were transfected transiently using Lipofectamine 3000 (Invitrogen, Carlsbad, CA).…”

mRNA Expression and Activity of Nucleoside Transporters in Human Hepatoma HepaRG Cells

“…HGF treatment was next demonstrated to concomitantly induce ENT1 activity (Figure 9D). Finally, the Janus kinase (JAK) inhibitor ruxolitinib, which has previously been shown to reverse IL6-mediated suppression of the hepatic sinusoidal drug transporters sodium-taurocholate cotransporting polypeptide (NTCP/ SLC10A1 ), OATP1B1 and OCT1 [37], also hindered IL6-mediated down-regulation of CNT1 (Figure 9E) and ENT2 (Figure 9F). By contrast, it failed to block IL1β-mediated suppression of CNT1 (Figure 9E) and ENT2 (Figure 9F) mRNA expression.…”