Abstract
The glutamatergic neurotransmitter system is involved in important neurophysiological processes and thus constitutes a promising target for the treatment of neurological diseases. The two ionotropic glutamate receptor agonists kainic acid (KA) and dihydrokainic acid (DHK) have been used as research tools in various in vivo central nervous system disease models in rodents, as well as being templates in the design of novel ligands affecting the glutamatergic system. Both molecules are highly polar but yet capable of crossing the blood–brain barrier (BBB). We used an in situ rat brain perfusion technique to determine the brain uptake mechanism and permeability across the BBB. To determine KA and DHK concentrations in the rat brain, simple and rapid sample preparation and liquid chromatography mass spectrometer methods were developed. According to our results the BBB permeability of KA and DHK is low, 0.25 × 10−6 and 0.28 × 10−6 cm/s for KA and DHK, respectively. In addition, the brain uptake is mediated by passive diffusion, and not by active transport. Furthermore, the non-specific plasma and brain protein binding of KA and DHK was determined to be low, which means that the unbound drug volume of distribution in brain is also low. Therefore, even though the total KA and DHK concentrations in the brain are low after systemic dosing, the concentrations in the vicinity of the glutamate receptors are sufficient for their activation and thus the observed efficacy.
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Abbreviations
- BBB:
-
Blood–brain barrier
- P:
-
Brain permeability
- fu,homogenate :
-
Buffer to tissue concentration ratio
- CNS:
-
Central nervous system
- DHK:
-
Dihydrokainic acid
- ESI:
-
Electrospray ionization
- KA:
-
Kainic acid
- M6G:
-
Morphine 6-beta-d-glucuronide
- MRM:
-
Multiple reaction monitoring
- Oatp:
-
Organic anion transporting polypeptide
- PBS:
-
Phosphate buffered saline
- \({\text{x}}_{{{\text{G}}^{ - } }}\) :
-
Transporter for anionic amino acids
- fu,brain :
-
Unbound fraction in brain
- fu,plasma :
-
Unbound fraction in plasma
- QC:
-
Quality control
- RSD:
-
Relative standard deviation
- K in :
-
Unidirectional transfer constant
References
Huneau C, Benquet P, Dieuset G, Biraben A, Martin B, Wendling F (2013) Shape features of epileptic spikes are a marker of epileptogenesis in mice. Epilepsia 54:2219–2227. doi:10.1111/epi.12406
Vivash L, Gregoire MC, Bouilleret V, Berard A, Wimberley C, Binns D, Roselt P, Katsifis A, Myers DE, Hicks RJ, O’Brien TJ, Dedeurwaerdere S (2014) In vivo measurement of hippocampal GABAA/cBZR density with [18F]-flumazenil PET for the study of disease progression in an animal model of temporal lobe epilepsy. PLoS One 9:e86722. doi:10.1371/journal.pone.0086722
Luquin MR, Saldise L, Guillen J, Belzunegui S, San Sebastian W, Izal A, Garrido P, Vazquez M (2006) Does increased excitatory drive from the subthalamic nucleus contribute to dopaminergic neuronal death in parkinson’s disease? Exp Neurol 201:407–415. doi:10.1016/j.expneurol.2006.04.033
Miller BR, Dorner JL, Bunner KD, Gaither TW, Klein EL, Barton SJ, Rebec GV (2012) Up-regulation of GLT1 reverses the deficit in cortically evoked striatal ascorbate efflux in the R6/2 mouse model of Huntington’s disease. J Neurochem 121:629–638. doi:10.1111/j.1471-4159.2012.07691.x
Ramirez C, Tercero I, Pineda A, Burgos JS (2011) Simvastatin is the statin that most efficiently protects against kainate-induced excitotoxicity and memory impairment. J Alzheimer’s Dis 24:161–174. doi:10.3233/JAD-2010-101653
Bunch L, Krogsgaard-Larsen P (2009) Subtype selective kainic acid receptor agonists: discovery and approaches to rational design. Med Res Rev 29:3–28
Bunch L, Nielsen B, Jensen AA, Brauner-Osborne H (2006) Rational design and enantioselective synthesis of (1R,4S,5R,6S)-3-azabicyclo [3.3.0] octane-4, 6-dicarboxylic acid a novel inhibitor at human glutamate transporter subtypes 1, 2, and 3. J Med Chem 49:172–178. doi:10.1021/jm0508336
Larsen AM, Venskutonyte R, Valades EA, Nielsen B, Pickering DS, Bunch L (2011) Discovery of a new class of ionotropic glutamate receptor antagonists by the rational design of (2S,3R)-3-(3-carboxyphenyl)-pyrrolidine-2-carboxylic acid. ACS Chem Neurosci 2:107–114. doi:10.1021/cn100093f
Juknaite L, Venskutonyte R, Assaf Z, Faure S, Gefflaut T, Aitken DJ, Nielsen B, Gajhede M, Kastrup JS, Bunch L, Frydenvang K, Pickering DS (2012) Pharmacological and structural characterization of conformationally restricted (S)-glutamate analogues at ionotropic glutamate receptors. J Struct Biol 180:39–46. doi:10.1016/j.jsb.2012.07.001
Battaglia A, Bertoluzza A, Calbucci F, Eusebi V, Giorgianni P, Ricci R, Tosi R, Tugnoli V (1999) High-performance liquid chromatographic analysis of physiological amino acids in human brain tumors by pre-column derivatization with phenylisothiocyanate. J Chromatogr B Biomed Sci Appl 730:81–93
Zhang X, Zhao T, Cheng T, Liu X, Zhang H (2012) Rapid resolution chromatography (RRLC) analysis of amino acids using pre-column derivatization. J Chromatogr B 906:91–95
Gynther M, Laine K, Ropponen J, Leppanen J, Mannila A, Nevalainen T, Savolainen J, Jarvinen T, Rautio J (2008) Large neutral amino acid transporter enables brain drug delivery via prodrugs. J Med Chem 51:932–936. doi:10.1021/jm701175d
Kalliokoski A, Niemi M (2009) Impact of OATP transporters on pharmacokinetics. Br J Pharmacol 158:693–705. doi:10.1111/j.1476-5381.2009.00430
Smith QR (2000) Transport of glutamate and other amino acids at the blood–brain barrier. J Nutr 130:1016S–1022S
Hawkins RA (2009) The blood–brain barrier and glutamate. Am J Clin Nutr 90:867S–874S. doi:10.3945/ajcn.2009.27462BB
Fenstermacher J, Gross P, Sposito N, Acuff V, Pettersen S, Gruber K (1988) Structural and functional variations in capillary systems within the brain. Ann N Y Acad Sci 529:21–30. doi:10.1111/j.1749-6632.1988.tb51416
Kalvass JC, Maurer TS (2002) Influence of nonspecific brain and plasma binding on CNS exposure: implications for rational drug discovery. Biopharm Drug Dispos 23:327–338. doi:10.1002/bdd.325
Summerfield SG, Read K, Begley DJ, Obradovic T, Hidalgo IJ, Coggon S, Lewis AV, Porter RA, Jeffrey P (2007) Central nervous system drug disposition: the relationship between in situ brain permeability and brain free fraction. J Pharmacol Exp Ther 322:205–213. doi:10.1124/jpet.107.121525
Maucher Fuquay J, Muha N, Wang Z, Ramsdell JS (2012) Elimination kinetics of domoic acid from the brain and cerebrospinal fluid of the pregnant rat. Chem Res Toxicol 25:2805–2809
Smith QR (2000) Transport of glutamate and other amino acids at the blood–brain barrier. J Nutr 130:1016S–1022S
Ronaldson PT, Davis TP (2013) Targeted drug delivery to treat pain and cerebral hypoxia. Pharmacol Rev 65:291–314
Hammarlund-Udenaes M, Friden M, Syvanen S, Gupta A (2008) On the rate and extent of drug delivery to the brain. Pharm Res 25:1737–1750
Stain-Texier F, Boschi G, Sandouk P, Scherrmann JM (1999) Elevated concentrations of morphine 6-beta-d-glucuronide in brain extracellular fluid despite low blood–brain barrier permeability. Br J Pharmacol 128:917–924. doi:10.1038/sj.bjp.0702873
van Vliet EA, Otte WM, Gorter JA, Dijkhuizen RM, Wadman WJ (2014) Longitudinal assessment of blood–brain barrier leakage during epileptogenesis in rats. A quantitative MRI study. Neurobiol Dis 63:74–84
Bauer B, Hartz AM, Pekcec A, Toellner K, Miller DS, Potschka H (2008) Seizure-induced up-regulation of P-glycoprotein at the blood–brain barrier through glutamate and cyclooxygenase-2 signaling. Mol Pharmacol 73:1444–1453
Loscher W, Potschka H (2005) Blood–brain barrier active efflux transporters: ATP-binding cassette gene family. NeuroRx 2:86–98
Mahar Doan KM, Humphreys JE, Webster LO, Wring SA, Shampine LJ, Serabjit-Singh CJ, Adkison KK, Polli JW (2002) Passive permeability and P-glycoprotein-mediated efflux differentiate central nervous system (CNS) and non-CNS marketed drugs. J Pharmacol Exp Ther 303:1029–1037
Acknowledgments
This study was funded by GluTarget (http://www.glutarget.ku.dk/), The Finnish Cultural Foundation and The Saastamoinen Foundation. The authors wish to thank Heidi Nielsen for her excellent technical assistance.
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Gynther, M., Petsalo, A., Hansen, S.H. et al. Blood–Brain Barrier Permeability and Brain Uptake Mechanism of Kainic Acid and Dihydrokainic Acid. Neurochem Res 40, 542–549 (2015). https://doi.org/10.1007/s11064-014-1499-4
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DOI: https://doi.org/10.1007/s11064-014-1499-4