In vitro and in vivo evidence for active brain uptake of the GHB analogue HOCPCA by the monocarboxylate transporter subtype 1

Research output: Contribution to journalJournal articlepeer-review

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In vitro and in vivo evidence for active brain uptake of the GHB analogue HOCPCA by the monocarboxylate transporter subtype 1. / Thiesen, Louise; Kehler, Jan; Clausen, Rasmus P; Frølund, Bente; Bundgaard, Christoffer; Wellendorph, Petrine.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 354, No. 2, 2015, p. 166-174 .

Research output: Contribution to journalJournal articlepeer-review

Harvard

Thiesen, L, Kehler, J, Clausen, RP, Frølund, B, Bundgaard, C & Wellendorph, P 2015, 'In vitro and in vivo evidence for active brain uptake of the GHB analogue HOCPCA by the monocarboxylate transporter subtype 1', Journal of Pharmacology and Experimental Therapeutics, vol. 354, no. 2, pp. 166-174 . https://doi.org/10.1124/jpet.115.224543

APA

Thiesen, L., Kehler, J., Clausen, R. P., Frølund, B., Bundgaard, C., & Wellendorph, P. (2015). In vitro and in vivo evidence for active brain uptake of the GHB analogue HOCPCA by the monocarboxylate transporter subtype 1. Journal of Pharmacology and Experimental Therapeutics, 354(2), 166-174 . https://doi.org/10.1124/jpet.115.224543

Vancouver

Thiesen L, Kehler J, Clausen RP, Frølund B, Bundgaard C, Wellendorph P. In vitro and in vivo evidence for active brain uptake of the GHB analogue HOCPCA by the monocarboxylate transporter subtype 1. Journal of Pharmacology and Experimental Therapeutics. 2015;354(2):166-174 . https://doi.org/10.1124/jpet.115.224543

Author

Thiesen, Louise ; Kehler, Jan ; Clausen, Rasmus P ; Frølund, Bente ; Bundgaard, Christoffer ; Wellendorph, Petrine. / In vitro and in vivo evidence for active brain uptake of the GHB analogue HOCPCA by the monocarboxylate transporter subtype 1. In: Journal of Pharmacology and Experimental Therapeutics. 2015 ; Vol. 354, No. 2. pp. 166-174 .

Bibtex

@article{4c99d7913d75417682d203c310a58d01,
title = "In vitro and in vivo evidence for active brain uptake of the GHB analogue HOCPCA by the monocarboxylate transporter subtype 1",
abstract = "γ-Hydroxybutyric acid (GHB) is a recreational drug, a clinically prescribed drug in narcolepsy and alcohol dependence, and an endogenous substance which binds to both high and low affinity sites in the brain. For studying the molecular mechanisms and the biological role of the GHB high-affinity binding sites, ligands with high and specific affinity are essential. The conformationally restricted GHB analogue 3-hydroxycyclopent-1-enecarboxylic acid (HOCPCA) is one such compound. The objective of this study was to investigate the transport of HOCPCA across the blood-brain barrier in vitro and in vivo, and to investigate the hypothesis that HOCPCA, like GHB, is a substrate for the monocarboxylate transporters (MCTs). For in vitro uptake studies, MCT1, 2 and 4 were recombinantly expressed in Xenopus laevis oocytes and the previously reported radioligand [(3)H]HOCPCA was used (as substrate). HOCPCA inhibited the uptake of the endogenous MCT substrate L-[(14)C]lactate, and [(3)H]HOCPCA was shown to act as substrate for MCT1 and 2 (Km values in the low millimolar range). Introducing single point amino acid mutations into positions essential for MCT function supported that HOCPCA binds to the endogenous substrate pocket of MCTs. MCT1-mediated brain entry of HOCPCA (10 mg/kg s.c.) was further confirmed in vivo in mice by co-administration of increasing doses of the MCT inhibitor [(R)-5-(3-hydroxypyrrolidine-1-carbonyl)-1-isobutyl-3-methyl-6-(quinolin-4-ylmethyl)thieno[2,3-d]pyrimidine-2,4(1H,3H)-dione] (AR-C141990) which inhibited brain penetration of HOCPCA in a dose-dependent manner (ID50 = 4.6 mg/kg). Overall, our study provides evidence that MCT1 is an important brain entry site for HOCPCA, and qualifies for future in vivo studies with HOCPCA.",
author = "Louise Thiesen and Jan Kehler and Clausen, {Rasmus P} and Bente Fr{\o}lund and Christoffer Bundgaard and Petrine Wellendorph",
note = "The American Society for Pharmacology and Experimental Therapeutics.",
year = "2015",
doi = "10.1124/jpet.115.224543",
language = "English",
volume = "354",
pages = "166--174 ",
journal = "Journal of Pharmacology and Experimental Therapeutics",
issn = "0022-3565",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "2",

}

RIS

TY - JOUR

T1 - In vitro and in vivo evidence for active brain uptake of the GHB analogue HOCPCA by the monocarboxylate transporter subtype 1

AU - Thiesen, Louise

AU - Kehler, Jan

AU - Clausen, Rasmus P

AU - Frølund, Bente

AU - Bundgaard, Christoffer

AU - Wellendorph, Petrine

N1 - The American Society for Pharmacology and Experimental Therapeutics.

PY - 2015

Y1 - 2015

N2 - γ-Hydroxybutyric acid (GHB) is a recreational drug, a clinically prescribed drug in narcolepsy and alcohol dependence, and an endogenous substance which binds to both high and low affinity sites in the brain. For studying the molecular mechanisms and the biological role of the GHB high-affinity binding sites, ligands with high and specific affinity are essential. The conformationally restricted GHB analogue 3-hydroxycyclopent-1-enecarboxylic acid (HOCPCA) is one such compound. The objective of this study was to investigate the transport of HOCPCA across the blood-brain barrier in vitro and in vivo, and to investigate the hypothesis that HOCPCA, like GHB, is a substrate for the monocarboxylate transporters (MCTs). For in vitro uptake studies, MCT1, 2 and 4 were recombinantly expressed in Xenopus laevis oocytes and the previously reported radioligand [(3)H]HOCPCA was used (as substrate). HOCPCA inhibited the uptake of the endogenous MCT substrate L-[(14)C]lactate, and [(3)H]HOCPCA was shown to act as substrate for MCT1 and 2 (Km values in the low millimolar range). Introducing single point amino acid mutations into positions essential for MCT function supported that HOCPCA binds to the endogenous substrate pocket of MCTs. MCT1-mediated brain entry of HOCPCA (10 mg/kg s.c.) was further confirmed in vivo in mice by co-administration of increasing doses of the MCT inhibitor [(R)-5-(3-hydroxypyrrolidine-1-carbonyl)-1-isobutyl-3-methyl-6-(quinolin-4-ylmethyl)thieno[2,3-d]pyrimidine-2,4(1H,3H)-dione] (AR-C141990) which inhibited brain penetration of HOCPCA in a dose-dependent manner (ID50 = 4.6 mg/kg). Overall, our study provides evidence that MCT1 is an important brain entry site for HOCPCA, and qualifies for future in vivo studies with HOCPCA.

AB - γ-Hydroxybutyric acid (GHB) is a recreational drug, a clinically prescribed drug in narcolepsy and alcohol dependence, and an endogenous substance which binds to both high and low affinity sites in the brain. For studying the molecular mechanisms and the biological role of the GHB high-affinity binding sites, ligands with high and specific affinity are essential. The conformationally restricted GHB analogue 3-hydroxycyclopent-1-enecarboxylic acid (HOCPCA) is one such compound. The objective of this study was to investigate the transport of HOCPCA across the blood-brain barrier in vitro and in vivo, and to investigate the hypothesis that HOCPCA, like GHB, is a substrate for the monocarboxylate transporters (MCTs). For in vitro uptake studies, MCT1, 2 and 4 were recombinantly expressed in Xenopus laevis oocytes and the previously reported radioligand [(3)H]HOCPCA was used (as substrate). HOCPCA inhibited the uptake of the endogenous MCT substrate L-[(14)C]lactate, and [(3)H]HOCPCA was shown to act as substrate for MCT1 and 2 (Km values in the low millimolar range). Introducing single point amino acid mutations into positions essential for MCT function supported that HOCPCA binds to the endogenous substrate pocket of MCTs. MCT1-mediated brain entry of HOCPCA (10 mg/kg s.c.) was further confirmed in vivo in mice by co-administration of increasing doses of the MCT inhibitor [(R)-5-(3-hydroxypyrrolidine-1-carbonyl)-1-isobutyl-3-methyl-6-(quinolin-4-ylmethyl)thieno[2,3-d]pyrimidine-2,4(1H,3H)-dione] (AR-C141990) which inhibited brain penetration of HOCPCA in a dose-dependent manner (ID50 = 4.6 mg/kg). Overall, our study provides evidence that MCT1 is an important brain entry site for HOCPCA, and qualifies for future in vivo studies with HOCPCA.

U2 - 10.1124/jpet.115.224543

DO - 10.1124/jpet.115.224543

M3 - Journal article

C2 - 25986445

VL - 354

SP - 166

EP - 174

JO - Journal of Pharmacology and Experimental Therapeutics

JF - Journal of Pharmacology and Experimental Therapeutics

SN - 0022-3565

IS - 2

ER -

ID: 138314195