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Donepezil, Tacrine and $\alpha-Phenyl-n-tert-Butyl Nitrone$ (PBN) Inhibit Choline Transport by Conditionally Immortalized Rat Brain Capillary Endothelial Cell Lines (TR-BBB)  

Kang Young-Sook (College of Pharmacy, Sookmyung Women's University)
Lee Kyeong-Eun (College of Pharmacy, Sookmyung Women's University)
Lee Na-Young (College of Pharmacy, Sookmyung Women's University)
Terasaki Tetsuya (Department of Molecular Biopharmacy and Genetics, Graduate School of Pharmaceutical Sciences, Tohoku University)
Publication Information
Archives of Pharmacal Research / v.28, no.4, 2005 , pp. 443-450 More about this Journal
Abstract
In the present study, we have characterized the choline transport system and examined the influence of various amine drugs on the choline transporter using a conditionally immortalized rat brain capillary endothelial cell line (TR-BBB) in vitro. The cell-to-medium (C/M) ratio of $[^3{H}]choline$ in TR-BBB cells increased time-dependently. The initial uptake rate of $[^3{H}]choline$ was concentration-dependent with a Michaelis-Menten value, $K_{m}$, of $26.2\pm2.7{\mu}M$. The $[^3{H}]choline$ uptake into TR-BBB was $Na^{+}-independent$, but was membrane potential-dependent. The $[^3{H}]choline$ uptake was susceptible to inhibition by hemicholinium-3, and tetraethy-lammonium (TEA), which are organic cation transporter substrates. Also, the uptake of $[^3{H}]choline$ was competitively inhibited with $K_{i}$ values of $274 {\mu}M, 251 {\mu}M and 180 {\mu}M$ in the presence of donepezil hydrochloride, tacrine and $\alpha-phenyl-n-tert-butyl nitrone$ (PBN), respectively. These characteristics of choline transport are consistent with those of the organic cation transporter (OCT). OCT2 mRNA was expressed in TR-BBB cells, while the expression of OCT3 or choline transporter (CHT) was not detected. Accordingly, these results suggest that OCT2 is a candidate for choline transport at the BBB and may influence the BBB permeability of amine drugs.
Keywords
Donepezil; tacrine; PBN; Choline transport; Organic cation transporter; Blood­brain barrier; Rat brain capillary endothelial cell line;
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1 Galea, E. and Estrada, C., Ouabain-sensitive choline transport system in capillaries isolated from bovine brain. J. Neurochem., 59, 936-941 (1992)   DOI   PUBMED
2 Klein, J., Gonzalez, R. K., Koppen, A., and Loffelholz, K., Free choline and choline metabolites in rat brain and body fluids: Sensitive determination and implications for choline supply to the brain. Neurochem. Int., 22, 293-300 (1993)   DOI   ScienceOn
3 Koepsell, H., Gorboulev, V., and Amdt, P., Molecular pharmacology of organic cation transporters in kidney. J. Membr. Biol., 167, 103-117 (1999)   DOI   ScienceOn
4 Mori, S., Takanaga, H., Ohtsuki, S., Deguchi, T., Kang, Y. S., Hosoya, K., and Terasaki, T., Rat organic anion transporter 3 (rOAT3) is responsible for brain-to-blood efflux of homovanillic acid at the abluminal membrane of brain capillary endothelial cells. J. Cereb. Blood Flow Metabol., 23, 432-440 (2003)   DOI
5 Pardridge, W. M., Holy grails and in vitro blood-brain barrier models. Drug Discov. Today, 9, 258 (2004)   DOI   ScienceOn
6 Saito, H., Masuda, S., and Inui, K., Cloning and functional characterization of a novel rat organic anion transporter mediating basolateral uptake of methotrexate in the kidney. J. Biol. Chem., 271, 20719-20725 (1996)   DOI   ScienceOn
7 Terasaki, T., Ohtsuki, S., Hori, S., Takanaga, H., Nakashima, E., and Hosoya, K., New approaches to in vitro models of bloodbrain barrier drug transport. Drug Discov. Today, 20, 944-954 (2003)   DOI   ScienceOn
8 Zhao, Q., Pahlmark, K., Smith, M. L., and Siesjo, B. K., Delayed treatment with the spin trap $\alpha$-phenyl-n-tert-butyl nitrone (PBN) reduces infarct size following transient middle cerebral artery occlusion in rats. Acta Physiol. Scand., 152, 349-350 (1994)   DOI   ScienceOn
9 Pardridge, W. M., Introduction to the blood-brain barrier: Methodology, biology and pathology, Cambridge University Press, Cambridge, pp. 1-486, (1998)
10 Diamond, I., Choline metabolism in the central nervous system: the role of choline transport from plasma to brain. Neurology, 20, 382 (1970)
11 Pardridge, W. M., Blood-brain barrier drug targeting: the future of brain drug development. Mol. Intervent., 3, 90-105 (2003)   DOI   PUBMED   ScienceOn
12 Okuda, T., Haga, T., Kanai, Y., Endou, H., Ishihara, T., and Katsura, I., Identification and characterization of the highaffinity choline transporter. Nat. Neurosci., 3, 120-125 (2000)   DOI   ScienceOn
13 Gomez, C., Martin, C., Galea, E., and Estrada, C., Direct cytotoxicity of ethylcholine mustard aziridinium in cerebral microvascular endothelial cells. J. Neurochem., 60, 1534- 1539 (1993)   DOI   PUBMED
14 Grundemann, D., Gorboulev, V., Gambaryan, S., Veyhl, M., and Koepsell, H., Drug excretion mediated by a new prototype of polyspecific transporter. Nature, 372, 549-552 (1994)   DOI   PUBMED   ScienceOn
15 Yamaoka, K., Tanigawara, Y., Nakagawa, T., and Uno, T., A pharmacokinetic analysis program (MULTI) for microcomputer. J. Pharmacobio-Dyn., 4, 879-885 (1981)   DOI
16 Hartvig, P., Askmark, H., Aquilonius, S. M., Wiklund, L., and Lindstrom, B., Clinical pharmacokinetics of intravenous and oral 9-amino-1,2,3,4-tetrahydroacridine, tacrine. Eur. J. Clin. Pharmacol., 38, 259-263 (1990)   DOI   ScienceOn
17 Johansson, M., Hellstrom-Lindahl, E., and Nordberg, A., Steadystate pharmacokinetics of tacrine in long-term treatment of Alzheimer patients. Dementia, 7, 111-117 (1996)
18 Rho, J. P. and Lipson, L. G., Focus on donepezil: A reversible acetylcholinesterase inhibitor for the treatment of Alzheimer's disease. Formulary, 32, 677-678 (1997)
19 Telting-Diaz, M. and Lunte, C. E., Distribution of tacrine across the blood-brain barrier in awake, freely moving rats using in vivo microdialysis sampling. Pharm. Res., 10, 44-48 (1993)   DOI   ScienceOn
20 Kekuda, R., Pasad, P. D., Wu, X., Wang, H., Fei, Y. J., Leibach, F. H., and Gaapathy, V., Cloning and functional characterization of a potential-sensitive, polyspecific organic cation transporter (OCT3) most abundantly expressed in placenta. J. Biol. Chem., 273, 15971-15979 (1998)   DOI   ScienceOn
21 Allen, D. D., Lockman, P. R., Roder, K. E., Dwoskin, L. P., and Crooks, P. A., Active transport of high-affinity choline and nicotine analogs into the central nervous system by the blood-brain barrier choline transporter. J. Pharm. Exp. Ther., 304, 1268-1274 (2003)   DOI   ScienceOn
22 Allen, D. D. and Smith, Q. R., Characterization of the bloodbrain barrier choline transporter using the in situ rat brain perfusion technique. J. Neurochem., 76, 1032-1041 (2001)   DOI   ScienceOn
23 Sweet, D. H., Miller, D. S., and Pritchard, J. B., Ventricular choline transport. J. Biol. Chem., 276, 41611-41619 (2001)   DOI   ScienceOn
24 Wu, X., Kekuda, R., Huang, W., Fei, Y. J., Leibach, F. H., Chen, J., Conway, S. J., and Ganapathy, V., Identity of the organic cation transporter OCT3 as the extraneuronal monoamine transporter (uptake2) and evidence for the expression of the transporter in the brain. J. Biol. Chem., 273, 32776-32786 (1998)   DOI   ScienceOn
25 Gorboulev, V., Ulzheimer, J. C., Akhoundova, A., Ulzheimer, Teuber, I., Karbach, U., Quester, S., Baumann, C., Lang, F., Busch, A. E., and Koepsell, H., Cloning and characterization of two human polyspecific organic cation transporters. DNA Cell Biol., 16, 871-881 (1997)   DOI   ScienceOn
26 Metting, T. L., Burgio, D. E., Terry, A. V., Beach, J. W., Mccurdy, C. R., and Allen, D. D., Inhibition of brain choline uptake by isoarecolone and lobeline derivatives: implications for potential vector-mediated brain drug delivery. Neurosci. Lett., 258, 25-28 (1998)   DOI   ScienceOn
27 Cornford, E. M., Braun, L. D., and Oldendorf, W. H., Carrier mediated blood-brain barrier transport of choline and certain choline analogs. J. Neurochem., 30, 299-308 (1978)   DOI   PUBMED
28 Grundemann, D., Babin-Ebell, J., Martel, F., Ording, N., Schmidt, A., and Schomig, E., Primary structure and functional expression of the apical organic cation transporter from kidney epithelial LLC-PK1 cells. J. Biol. Chem., 272, 10408- 10413 (1997)   DOI   ScienceOn
29 Hosoya, K., Takashima, T., Tetsuka, K., Nagura, T., Ohtsuki, S., Takanaga, H., Ueda, M., Yanai, N., Obinata, M., and Terasaki, T., mRNA expression and transport characterization of conditionally immortalized rat brain capillary endothelial cell lines; a new in vitro BBB model for drug targeting. J. Drug Target, 8, 357-370 (2000)   DOI   ScienceOn
30 Sawada, N., Takanaga, H., Matsuo, H., Naito, M., Tsuruo, T., and Sawada, Y., Choline uptake by mouse brain capillary endothelial cells in culture. J. Pharm. Pharmacol., 51, 847- 852 (1999)   DOI
31 Parfitt, K. and Martindale, W., Martindale: the complete drug reference. 32nd ed. Pharmaceutical Press, London, pp. 1391- 1392 (1999)
32 Friedrich, A., George, R. L., Bridges, C. C., Prasad, P. D., and Ganapathy, V., Transport of choline and its relationship to the expression of the organic cation transporters in a rat brain microvessel endothelial cell line (RBE4). Biochim. Biophys. Acta, 1512, 299-307 (2001)   DOI   ScienceOn
33 Tiseo, P. J., Rogers, S. L., and Friedhoff, L. T., Pharmacokinetic and pharmacodynamic profile of donepezil HCl following evening administration. Br. J. Clin. Pharmacol., 46 Suppl 1, 13-18 (1998)   DOI   PUBMED
34 Matsui, K., Mishima, M., Nagai, Y., Yuzuriha, T., and Yoshimura, T., Absorption, distribution, metabolism, and excretion of donepezil (Aricept) after a single oral administration to rat. Drug Metab. Dispos., 27, 1406-1414 (1999)
35 Tamai, I. and Tsuji, A., Transporter-mediated permeation of drugs across the blood-brain barrier. J. Pharm. Sci., 89, 1371-1388 (2000)   DOI   ScienceOn
36 Pardridge, W. M. and Oldendorf, W. H., Transport of metabolic substrates through the blood-brain barrier. J. Neurochem., 28, 5-12 (1977)   DOI   PUBMED
37 MaNally, W. P., Pool, W. F., Sinz, M. W., Dehart, P., Ortwine, D. F., Huang, C. C., Chang, T., and Woolf, T. F., Distribution of tacrine and metabolites in rat brain and plasma after singleand multiple-dose regimens; Evidence for accumulation of tacrine in brain tissue. Drug Metab. Dispos., 24, 628-633 (1996)
38 Kang, Y. S., Terasaki, T., Ohnishi, T., and Tsuji, A., In vivo and in vitro evidence for a common carrier mediated transport of choline and basic drugs through the blood-brain barrier. J. Pharmacobio-Dyn., 13, 353-360 (1990)   DOI
39 Grundemann, D., Koster, S., Kiefer, N., Breidert, T., Engelhardt, M., Spitzenberger, F., Obermuller, N., and Schomig, E., Transport of monoamine transmitters by the organic cation transporter type 2, OCT2. J. Biol. Chem., 273, 30915-30920 (1998)   DOI   ScienceOn
40 Knecht, K. T. and Mason, R. P., In vivo spin trapping of xenobiotic free radical metabolites. Arch. Biochem. Biophys., 303, 185-194 (1993)   DOI   ScienceOn
41 Wu, X., Huang, W., Prasak, P. D., Seth, P., Rajan, D. P., Leibach, F. H., Chen, J., Conway, S. J., and Ganapathy, V., Functional characteristics and tissue distribution pattern of organic cation transporter 2 (OCTN2), an organic cation/ carnitine transporter. J. Pharm. Exp. Ther., 290, 1482-1492 (1999)   PUBMED