Evaluating Drug–Drug Interaction Risk Associated with Peptide Analogs Using advanced In Vitro Systems
Rune Aa. Nørgaard, D. Bhatt, Erkka Järvinen, Tore Bjerregaard Stage, Charlotte Gabel‐Jensen, Aleksandra Galetin, Carolina Säll
Abstract
Drug-drug interaction (DDI) assessment of therapeutic peptides is an evolving area. The industry generally follows DDI guidelines for small molecules, but the translation of data generated with commonly used <i>in vitro</i> systems to <i>in vivo</i> is sparse. In the current study, we investigated the ability of advanced human hepatocyte <i>in vitro </i>systems namely HepatoPac, spheroids, and Liver-on-a-chip to assess potential changes in regulation of CYP1A2, CYP2B6, CYP3A4, <i>SLCO1B1</i> and <i>ABCC2</i> in the presence of selected therapeutic peptides, proteins, and small molecules. The peptide NN1177, a glucagon and GLP-1 receptor co-agonist, did not suppress mRNA expression or activity of CYP1A2, CYP2B6, and CYP3A4 in HepatoPac, spheroids, or Liver-on-a-chip; these findings were in contrast to the data obtained in sandwich cultured hepatocytes. No effect of NN1177 on <i>SLCO1B1</i> and <i>ABCC2</i> mRNA was observed in any of the complex systems. The induction magnitude differed across the systems (e.g., rifampicin induction of <i>CYP3A4</i> mRNA ranged from 2.8-fold in spheroids to 81.2-fold in Liver-on-a-chip). Small molecules, obeticholic acid and abemaciclib, showed varying responses in HepatoPac, spheroids and Liver-on-a-chip, indicating a need for EC<sub>50</sub> determinations to fully assess translatability data. HepatoPac, the most extensively investigated in this study (3 donors), showed high potential to investigate DDIs associated with CYP regulation by therapeutic peptides. Spheroids and Liver-on-a-chip were only assessed in one hepatocyte donor and further evaluations are required to confirm their potential. This study establishes an excellent foundation towards the establishment of more clinically-relevant <i>in vitro </i>tools for evaluation of potential DDIs with therapeutic peptides. <b>Significance Statement</b> At present, there are no guidelines for drug-drug interaction (DDI) assessment of therapeutic peptides. Existing <i>in vitro</i> methods recommended for assessing small molecule DDIs do not appear to translate well for peptide drugs, complicating drug development for these moieties. Here, we establish evidence that complex cellular systems have potential to be used as more clinically-relevant tools for the <i>in vitro</i> DDI evaluation of therapeutic peptides.