Human hepatocytes and cytochrome P450‐selective inhibitors predict variability in human drug exposure more accurately than human recombinant P450s

Abstract

The results suggest that variability in human drug exposure due to DDI and enzyme polymorphism can be accurately predicted using fm from human hepatocytes and CYP-selective inhibitors. This approach can be efficiently applied in drug discovery to aid optimization of candidate drugs with a favourable metabolic elimination profile and limited variability in patients.

In Vitro Characterization of Ertugliflozin Metabolism by UDP-Glucuronosyltransferase and Cytochrome P450 Enzymes

“…Orthogonal ISEF-adjusted scaling of rhCYP3A4 and CYP2C8 CLint, suggested a lessor role of CYP2C8 (fm 0.01) relative to CYP3A4 (fm 0.99). The increased relative contribution of CYP3A is apparently due to the tendency of ISEF-adjusted rhCYP CLint to overestimate CYP3A-mediated CL relative to other CYP enzymes (Chen et al, 2011;Umehara et al, 2017;Lindmark et al, 2018).…”

In vitro and in vivo methods to assess pharmacokinetic drug– drug interactions in drug discovery and development

“…HHEP is a more physiologically relevant system compared with the other enzyme sources (e.g., HLM or recombinant enzymes) and has a complete complement of Phase I/II enzymes and cofactors. Reaction phenotyping data with HHEP have been shown to be able to predict human in vivo DDI with high accuracy (Lindmark, Lundahl, Kanebratt, Andersson, & Isin, ). Selective inhibitors that are used for CYP reaction phenotyping in HHEP have been reported (Yang, Atkinson, & Di, ).…”

“…These include substrate dependence of correction factors, non‐physiological conditions with excess cytochrome b5 and P450 reductase, lack of cooperativity and protein–protein interactions (Chen, Liu, Nguyen, & Fretland, ; Di, ; Konopnicki et al, ; Siu & Lai, ). The accuracy for predicting human DDI using f m from human recombinant enzymes is not as reliable as using HHEP (Lindmark et al, ). In a comparison study of CYP3A and CYP2D6 substrates using hrCYPs and HHEP, the f m values obtained from HHEP gave an accurate prediction of human DDI AUC changes, while the hrCYPs were not as accurate and tended to overestimate the CYP3A contribution (Lindmark et al, ).…”

“…The accuracy for predicting human DDI using f m from human recombinant enzymes is not as reliable as using HHEP (Lindmark et al, ). In a comparison study of CYP3A and CYP2D6 substrates using hrCYPs and HHEP, the f m values obtained from HHEP gave an accurate prediction of human DDI AUC changes, while the hrCYPs were not as accurate and tended to overestimate the CYP3A contribution (Lindmark et al, ). This is an issue particularly for low clearance compounds, for which CYP3A is identified as the only pathway in many cases due to no turnover in the other hrCYPs.…”

(–)-N-3-Benzylphenobarbital Is Superior to Omeprazole and (+)-N-3-Benzylnirvanol as a CYP2C19 Inhibitor in Suspended Human Hepatocytes

“…However, there are several disadvantages to using recombinant enzymes, namely, (i) the evaluation of the cooperation between enzymes is impossible, (ii) a relative activity factor is needed for scaling to in vivo, (iii) enzymes are removed from their natural intracellular environment and (iv) the addition of cofactors is required (Brandon et al, 2003). Human liver microsomes are frequently used because of their low cost, high availability, ease of use, and high throughput (Lindmark et al, 2018). They represent a more advanced experimental model than recombinant enzymes, despite the fact that they also require the addition of cofactors and that they lack transporters or cell membranes as well as several key drug-metabolizing enzymes (e.g.…”