Expansion of Knowledge on OCT1 Variant Activity In Vitro and In Vivo Using Oct1/2−/− Mice

Bridget L Morse,Philip C Smith,John K Fallon,Lisa Hong Chen,Kathleen M Hillgren,John T Catlow

doi:10.3389/fphar.2021.631793

Abstract

The role of organic cation transporter 1 (OCT1) in humans is gaining attention as data emerges regarding its role in physiology, drug exposure, and drug response. OCT1 variants with decreased in vitro function correlate well with altered exposure of multiple OCT1 substrates in variant carriers. In the current research, we investigate mechanisms behind activity of OCT1 variants in vitro by generating cell lines expressing known OCT1 variants and quantifying membrane OCT1 protein expression with corresponding OCT1 activity and kinetics. Oct knockout mice have provided additional insight into the role of Oct1 in the liver and have reproduced effects of altered OCT1 activity observed in the clinic. To assess the complex effect of Oct1 depletion on pharmacokinetics of prodrug proguanil and its active moiety cycloguanil, both of which are OCT1 substrates, Oct1/2−/− mice were used. Decreased membrane expression of OCT1 was demonstrated for all variant cell lines, although activity was substrate-dependent, as reported previously. Lack of change in activity for OCT1*2 resulted in increased intrinsic activity per pmol of OCT1 protein, particularly for sumatriptan but also for proguanil and cycloguanil. Similar to that reported in humans with decreased OCT1 function, systemic exposure of proguanil was minimally affected in Oct1/2−/− mice. However, proguanil liver partitioning and exposure decreased. Cycloguanil exposure decreased following proguanil administration in Oct1/2−/− mice, as did the systemic metabolite:parent ratio. When administered directly, systemic exposure of cycloguanil decreased slightly; however liver partitioning and exposure were decreased in Oct1/2−/− mice. Unexpectedly, following proguanil administration, the metabolite ratio in the liver changed only minimally, and liver partitioning of cycloguanil was affected in Oct1/2−/− mice to a lesser extent following proguanil administration than direct administration of cycloguanil. In conclusion, these in vitro and in vivo data offer additional complexity in understanding mechanisms of OCT1 variant activity as well as the effects of these variants in vivo. From cell lines, it is apparent that intrinsic activity is not directly related to OCT1 membrane expression. Additionally, in situations with a more complicated role of OCT1 in drug pharmacokinetics there is difficulty translating in vivo impact simply from intrinsic activity from cellular data.

Highlights

In humans, organic cation transporter 1 (OCT1) is localized in the liver and intestine, organs pertaining to drug absorption, distribution, metabolism and excretion (Drozdzik et al, 2019)
A prominent example is the effect of OCT1 activity on the metabolite ratio for cycloguanil:proguanil, in which the authors were able to demonstrate a continuous correlation of in vitro activity to the relationship observed in vivo (Matthaei et al, 2019)
The uptake of known clinical OCT1 substrates in cell lines expressing wildtype and variant OCT1 protein are shown in Figure 1; uptake values are shown both before (A) and after (B) normalizing for measured membrane expressed OCT1 protein

Summary

INTRODUCTION

Organic cation transporter 1 (OCT1) is localized in the liver and intestine, organs pertaining to drug absorption, distribution, metabolism and excretion (Drozdzik et al, 2019). As many therapeutic agents are demonstrated OCT1 substrates (Koepsell, 2020), the potential exists for a role of OCT1mediated transport on the disposition of therapeutic drugs This prospect was propagated by the identification of OCT1 variants with varying degrees of transport activity impacting cellular exposure, initially on metformin uptake but followed by sumatriptan, fenoterol, proguanil, ranitidine and others (Shu et al, 2007; Matthaei et al, 2015; Meyer et al, 2017; Tzvetkov et al, 2018; Matthaei et al, 2019; Jensen et al, 2020). We previously assessed Oct1/2−/− mice as a model for reproducing or predicting the effect of OCT1 variants in the clinic In these studies, hepatic clearance of sumatriptan and fenoterol was significantly decreased, and the change in oral and IV clearance was similar to that reported in human carriers of OCT1 null variants (Morse et al, 2020). We use the model to assess the liver exposure changes in these agents, as this is a site of action and may add to hypotheses on liver exposure of these therapeutic agents in patients with decreased OCT1 activity

MATERIALS AND METHODS

RESULTS

DISCUSSION

ETHICS STATEMENT