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In Vivo and In Vitro Evidence for Brain Uptake of 4-Phenylbutyrate by the Monocarboxylate Transporter 1 (MCT1)

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Abstract

Purpose

4-Phenylbutyrate (4-PBA) is expected to be a potential therapeutic for several neurodegenerative diseases. These activities require 4-PBA transport into the brain across the blood-brain barrier (BBB). The objective of the present study was to characterize the brain transport mechanism of 4-PBA through the BBB.

Methods

The brain transport of 4-PBA across the BBB was investigated following intravenous (IV) injection and internal carotid artery perfusion (ICAP) in vivo. The mechanism of transport was examined using TR-BBB cells, an in vitro model of the BBB.

Results

The volume of distribution (VD) of 4-PBA by rat brain was about 7-fold greater than that of sucrose, a BBB impermeable vascular space marker, suggesting the blood-to-brain transport of 4-PBA through the BBB in the physiological state. [14C]4-PBA uptake by TR-BBB cells showed time-, pH- and concentration-dependence with a K m of 13.4 mM at pH 7.4 and 3.22 mM at pH 6.0. The uptake was Na+ independent, and was significantly inhibited by alpha-cyano-4-hydroxycinnamate (a typical inhibitor for monocarboxylate transport), endogenous monocarboxylate compounds and monocarboxylic drugs. Lactate and valproate competitively inhibited [14C]4-PBA uptake with K i value of 13.5 mM and 7.47 mM, respectively. These results indicate the role of monocarboxylate transporters (MCTs) in 4-PBA transport into the brain at the BBB. TR-BBB cells expressed mRNA of rMCT1, 2, and 4, especially, rMCT1 showed high mRNA expression level. In addition, [14C]4-PBA uptake was inhibited by rMCT1 specific small interfering RNA.

Conclusion

The transport mechanism of 4-PBA from blood to brain across the BBB likely involves MCT1.

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Abbreviations

4- PBA:

4-Phenylbutyrate

AD:

Alzheimer’s disease

ALS:

Amyotrophic lateral sclerosis

BBB:

Blood-brain barrier

CHC:

Alpha-cyano-4-hydroxycinnamate

CSF:

Cerebrospinal fluid

ECF:

Extracellular fluid

ER:

Endoplasmic reticulum

FCCP:

Carbonyl cyanide p-trifluoromethoxyphenylhydrazone

GHB:

γ–Hydroxybutyrate

HD:

Huntington disease

HDAC:

Histone deacetylase

ICAP:

Internal carotid artery perfusion

ID:

Injected dose

IV:

Intravenous

KHB:

Krebs–Henseleit buffer

K i :

Inhibition constant

K m :

Michaelis-Menten constant

K ns :

Nonsaturable uptake clearance

MCT:

Monocarboxylate transporter

NMDG:

N-methyl-D-glucamine

PAH:

p-Aminohippurate

PD:

Parkinson’s disease

PS:

BBB permeability-surface area

SD:

Sprague-Dawley

SMCT:

Na+ coupled monocarboxylate transporter

TAUT:

Taurine transporter

TR-BBB cells:

Conditionally immortalized rat brain capillary endothelial cell line

Vd :

Terminal brain/plasma ratio

VD :

Volume of distribution

V max :

Maximum rate of uptake

VPA:

Valproate

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ACKNOWLEDGMENTS AND DISCLOSURES

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (No. 2011-0030701) and the SRC Research Center for Women’s Diseases of Sookmyung Women’s University (No. 3-1103-0021).

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  1. College of Pharmacy and Research Center for Cell Fate Control, Sookmyung Women’s University, 52, Hyochangwon-gil, Yongsan-gu, Seoul, 04310, South Korea

    Na-Young Lee & Young-Sook Kang

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Correspondence to Young-Sook Kang.

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Lee, NY., Kang, YS. In Vivo and In Vitro Evidence for Brain Uptake of 4-Phenylbutyrate by the Monocarboxylate Transporter 1 (MCT1). Pharm Res 33, 1711–1722 (2016). https://doi.org/10.1007/s11095-016-1912-6

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  • DOI: https://doi.org/10.1007/s11095-016-1912-6

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