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Participation of Monocarboxylate Transporter 8, But Not P-Glycoprotein, in Carrier-Mediated Cerebral Elimination of Phenytoin across the Blood-Brain Barrier

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Abstract

Purpose

In this study, we investigated in detail the transport of phenytoin across the blood-brain barrier (BBB) to identify the transporter(s) involved in BBB-mediated phenytoin efflux from the brain.

Methods

We evaluated the brain-to-blood efflux transport of phenytoin in vivo by determining the brain efflux index (BEI) and uptake in brain slices. We additionally conducted brain perfusion experiments and BEI studies in P-glycoprotein (P-gp)-deficient mice. In addition, we determined the mRNA expression of monocarboxylate transporter (MCT) in isolated brain capillaries and performed phenytoin uptake studies in MCT-expressing Xenopus oocytes.

Results

[14C]Phenytoin brain efflux was time-dependent with a half-life of 17 min in rats and 31 min in mice. Intracerebral pre-administration of unlabeled phenytoin attenuated BBB-mediated phenytoin efflux transport, suggesting carrier-mediated phenytoin efflux transport across the BBB. Pre-administration of P-gp substrates in rats and genetic P-gp deficiency in mice did not affect BBB-mediated phenytoin efflux transport. In contrast, pre-administration of MCT8 inhibitors attenuated phenytoin efflux. Moreover, rat MCT8-expressing Xenopus oocytes exhibited [14C]phenytoin uptake, which was inhibited by unlabeled phenytoin.

Conclusion

Our data suggest that MCT8 at the BBB participates in phenytoin efflux transport from the brain to the blood.

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Abbreviations

ABC:

ATP-binding cassette

BBB:

Blood-brain barrier

BEI:

Brain efflux index

BCRP:

Breast cancer resistance protein

BSP:

Bromosulfophthalein

CEF:

Cerebral extracellular fluid

cDNA:

Complementary DNA

cRNA:

Complementary RNA

CYP:

Cytochrome P450

DHEAS:

Dehydroepiandrosterone-3-sulfate

HEPES:

4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid

MCT:

Monocarboxylate transporter

MRP:

Multidrug resistance-associated protein

n :

Group sizes

P-gp:

P-glycoprotein

S.D.:

Standard deviation

SNP:

Single nucleotide polymorphism

X :

Xenopus

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ACKNOWLEDGMENTS AND DISCLOSURES

The authors thank Mr. Kosuke Tajima (University of Toyama) for helping with the mRNA expression analysis for MCTs. This research was supported by The Mochida Memorial Foundation for Medical and Pharmaceutical Research, The Research Foundation for Pharmaceutical Sciences, and Japan Society for the Promotion of Science (JSPS) KAKENHI [Grant Numbers JP16K08365 (to S.A.), JP16H05110 (to K.H.), and JP19K07160 (to S.A.)]. This project was also supported by Grant 1R01NS079507 from the U.S. National Institutes of Health (NIH)/National Institute of Neurological Disorders and Stroke (to B.B.). The content is solely the responsibility of the authors, and it does not necessarily represent the official views of the National Institute of Neurological Disorders and Stroke or the NIH. All authors declare no conflict of interest.

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Authors and Affiliations

  1. Department of Pharmaceutics, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan

    Ryuta Jomura, Shin-ichi Akanuma, Yukiko Yoshida, Yoshiyuki Kubo & Ken-ichi Hosoya

  2. Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, 800 S Limestone, Lexington, Kentucky, 40536-0230, USA

    Shin-ichi Akanuma & Björn Bauer

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  1. Ryuta Jomura
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Contributions

R.J., S.A., B.B., and Y.Y. performed the experiments; R.J., S.A., B.B., Y.K., and H.K. designed the experiments and analyzed the data.; and R.J. and S.A. wrote the manuscript. All authors read the final version of the manuscript.

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Correspondence to Shin-ichi Akanuma.

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Jomura, R., Akanuma, Si., Bauer, B. et al. Participation of Monocarboxylate Transporter 8, But Not P-Glycoprotein, in Carrier-Mediated Cerebral Elimination of Phenytoin across the Blood-Brain Barrier. Pharm Res 38, 113–125 (2021). https://doi.org/10.1007/s11095-021-03003-1

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