Pharmaceutical Research

, Volume 30, Issue 10, pp 2523–2537 | Cite as

Design, Synthesis and Brain Uptake of LAT1-Targeted Amino Acid Prodrugs of Dopamine

  • Lauri PeuraEmail author
  • Kalle Malmioja
  • Kristiina Huttunen
  • Jukka Leppänen
  • Miia Hämäläinen
  • Markus M. Forsberg
  • Jarkko Rautio
  • Krista Laine
Research Paper



Drug delivery to the brain is impeded by the blood-brain barrier (BBB). Here, we attempted to enhance the brain uptake of cationic dopamine by utilizing the large amino acid transporter 1 (LAT1) at the BBB by prodrug approach.


Three amino acid prodrugs of dopamine were synthesized and their prodrug properties were examined in vitro. Their LAT1-binding and BBB-permeation were studied using the in situ rat brain perfusion technique. The brain uptake after intravenous administration and the dopamine-releasing ability in the rat striatum after intraperitoneal administration were also determined for the most promising prodrug.


All prodrugs underwent adequate cleavage in rat tissue homogenates. The prodrug with phenylalanine derivative as the promoiety had both higher affinity for LAT1 and better brain uptake properties than those with an alkyl amino acid -mimicking promoiety. The phenylalanine prodrug was taken up into the brain after intravenous injection but after intraperitoneal injection the prodrug did not elevate striatal dopamine concentrations above those achieved by corresponding L-dopa treatment.


These results indicate that attachment of phenylalanine to a cationic drug via an amide bond from the meta-position of its aromatic ring could be highly applicable in prodrug design for LAT1-mediated CNS-delivery of not only anionic but also cationic polar drugs.


blood-brain barrier brain drug delivery dopamine large amino acid transporter 1 prodrugs 



aromatic L-amino acid decarboxylase


area under the curve


blood-brain barrier




central nervous system


catechol-O-methyl transferase




glucose transporter 1




large neutral amino acid transporter 1


lower limit of quantification


brain permeability-surface area


ascorbic acid transporter


topological polar surface area



Lauri Peura, Kalle Malmioja, Jarkko Rautio and Krista Laine share equal contribution to this work.

The authors would like to express their profound gratitude to laboratory technicians Helly Rissanen and Jaana Leskinen for their skillful assistance. Marko Lehtonen, M.Sc., is also acknowledged for his skillful help in the HPLC-EC-analytics, Tiina Kääriäinen, Ph.D., for her skillful help in the animal studies and Ewen MacDonald, Ph.D., for refining the English of this paper. We also thank Henna Härkönen, M.Sc., for the polar surface area calculations. The work was financially supported by the Graduate School of Pharmaceutical Research, the Academy of Finland (# 132637), the Orion-Farmos foundation, the Finnish Pharmaceutical Society, the Finnish Parkinson Foundation and the Emil Aaltonen Foundation.


  1. 1.
    Begley DJ. Delivery of therapeutic agents to the central nervous system: the problems and the possibilities. Pharmacol Ther. 2004;104(1):29–45.PubMedCrossRefGoogle Scholar
  2. 2.
    Pardridge WM. Crossing the blood–brain barrier: are we getting it right? Drug Discov Today. 2001;6(1):1–2.PubMedCrossRefGoogle Scholar
  3. 3.
    Wager TT, Hou X, Verhoest PR, Villalobos A. Moving beyond rules: the development of a central nervous system multiparameter optimization (CNS MPO) approach to enable alignment of druglike properties. ACS Chem Neurosci. 2010;1(6):435–49.PubMedCrossRefGoogle Scholar
  4. 4.
    Pavan B, Dalpiaz A. Prodrugs and endogenous transporters: are they suitable tools for drug targeting into the central nervous system? Curr Pharm Des. 2011;17(32):3560–76.PubMedCrossRefGoogle Scholar
  5. 5.
    Gynther M, Ropponen J, Laine K, Leppänen J, Haapakoski P, Peura L, et al. Glucose promoiety enables glucose transporter mediated brain uptake of ketoprofen and indomethacin prodrugs in rats. J Med Chem. 2009;52(10):3348–53.PubMedCrossRefGoogle Scholar
  6. 6.
    Bonina F, Puglia C, Rimoli MG, Melisi D, Boatto G, Nieddu M, et al. Glycosyl derivatives of dopamine and L-dopa as anti-Parkinson prodrugs: Synthesis, pharmacological activity and in vitro stability studies. J Drug Target. 2003;11(1):25–36.PubMedGoogle Scholar
  7. 7.
    Gynther M, Jalkanen A, Lehtonen M, Forsberg M, Laine K, Ropponen J, et al. Brain uptake of ketoprofen-lysine prodrug in rats. Int J Pharm. 2010;399(1–2):121–8.PubMedCrossRefGoogle Scholar
  8. 8.
    Gynther M, Laine K, Ropponen J, Leppänen J, Mannila A, Nevalainen T, et al. Large neutral amino acid transporter enables brain drug delivery via prodrugs. J Med Chem. 2008;51(4):932–6.PubMedCrossRefGoogle Scholar
  9. 9.
    Hokari M, Wu H, Schwarcz R, Smith QR. Facilitated brain uptake of 4-chlorokynurenine and conversion to 7-chlorokynurenic acid. Neuroreport. 1997;8(1):15–8.CrossRefGoogle Scholar
  10. 10.
    Killian DM, Chikhale PJ. A bioreversible prodrug approach designed to shift mechanism of brain uptake for amino-acid-containing anticancer agents. J Neurochem. 2001;76(4):966–74.PubMedCrossRefGoogle Scholar
  11. 11.
    Bonina FP, Arenare L, Palagiano F, Saija A, Nava F, Trombetta D, et al. Synthesis, stability, and pharmacological evaluation of nipecotic acid prodrugs. J Pharm Sci. 1999;88(5):561–7.PubMedCrossRefGoogle Scholar
  12. 12.
    Balakrishnan A, Jain-Vakkalagadda B, Yang C, Pal D, Mitra AK. Carrier mediated uptake of L-tyrosine and its competitive inhibition by model tyrosine linked compounds in a rabbit corneal cell line (SIRC)—strategy for the design of transporter/receptor targeted prodrugs. Int J Pharm. 2002;247(1–2):115–25.PubMedCrossRefGoogle Scholar
  13. 13.
    Peura L, Malmioja K, Laine K, Leppänen J, Gynther M, Isotalo A, et al. Large amino acid transporter 1 (LAT1) prodrugs of valproic acid: new prodrug design ideas for central nervous system delivery. Mol Pharm. 2011;8(5):1857–66.PubMedCrossRefGoogle Scholar
  14. 14.
    Gomes P, Soares-Da-Silva P. L-DOPA transport properties in an immortalised cell line of rat capillary cerebral endothelial cells, RBE 4. Brain Res. 1999;829(1–2):143–50.PubMedCrossRefGoogle Scholar
  15. 15.
    Nutt JG. Pharmacokinetics and pharmacodynamics of levodopa. Mov Disord. 2008;23(S3):S580–4.PubMedCrossRefGoogle Scholar
  16. 16.
    Olanow C, Obeso J, Stocchi F. Drug insight: continuous dopaminergic stimulation in the treatment of Parkinson’s disease. Nat Clin Pract Neurol. 2006;2(7):382–92.PubMedCrossRefGoogle Scholar
  17. 17.
    Chemuturi NV, Donovan MD. Role of organic cation transporters in dopamine uptake across olfactory and nasal respiratory tissues. Mol Pharm. 2007;4(6):936–42.PubMedCrossRefGoogle Scholar
  18. 18.
    Borgman RJ, McPhillips JJ, Stitzel RE, Goodman IJ. Synthesis and pharmacology of centrally acting dopamine derivatives and analogs in relation to Parkinson’s disease. J Med Chem. 1973;16(6):630–3.PubMedCrossRefGoogle Scholar
  19. 19.
    Bodor N, Farag HH, Brewster III ME. Site-specific, sustained release of drugs to the brain. Science. 1981;214(4527):1370–2.PubMedCrossRefGoogle Scholar
  20. 20.
    Bodor N, Farag HH. Improved delivery through biological membranes. 11. A redox chemical drug-delivery system and its use for brain-specific delivery of phenylethylamine. J Med Chem. 1983;26(3):313–8.PubMedCrossRefGoogle Scholar
  21. 21.
    Simpkins JW, Bodor N, Enz A. Direct evidence for brain-specific release of dopamine from a redox delivery system. J Pharm Sci. 1985;74(10):1033–6.PubMedCrossRefGoogle Scholar
  22. 22.
    Carelli V, Liberatore F, Scipione L, Impicciatore M, Barocelli E, Cardellini M, et al. New systems for the specific delivery and sustained release of dopamine to the brain. J Control Release. 1996;42(3):209–16.CrossRefGoogle Scholar
  23. 23.
    Denora N, Laquintana V, Lopedota A, Serra M, Dazzi L, Biggio G, et al. Novel L-Dopa and dopamine prodrugs containing a 2-phenyl-imidazopyridine moiety. Pharm Res. 2007;24(7):1309–24.PubMedCrossRefGoogle Scholar
  24. 24.
    Fernández C, Nieto O, Fontenla JA, Rivas E, De Ceballos ML, Fernandez-Mayoralas A. Synthesis of glycosyl derivatives as dopamine prodrugs: interaction with glucose carrier GLUT-1. Org Biomol Chem. 2003;1(5):767–71.PubMedCrossRefGoogle Scholar
  25. 25.
    Fernández C, Nieto O, Rivas E, Montenegro G, Fontenla JA, Fernández-Mayoralas A. Synthesis and biological studies of glycosyl dopamine derivatives as potential antiparkinsonian agents. Carbohydr Res. 2000;327(4):353–65.PubMedCrossRefGoogle Scholar
  26. 26.
    More SS, Vince R. Design, synthesis and biological evaluation of glutathione peptidomimetics as components of anti-Parkinson prodrugs. J Med Chem. 2008;51(15):4581–8.PubMedCrossRefGoogle Scholar
  27. 27.
    Dent III WH, Erickson WR, Fields SC, Parker MH, Tromiczak EG. 9-BBN: an amino acid protecting group for functionalization of amino acid side chains in organic solvents. Org Lett. 2002;4(8):1249–51.PubMedCrossRefGoogle Scholar
  28. 28.
    Ertl P, Rohde B, Selzer P. Fast calculation of molecular polar surface area as a sum of fragment-based contributions and its application to the prediction of drug transport properties. J Med Chem. 2000;43(20):3714–7.PubMedCrossRefGoogle Scholar
  29. 29.
    Takasato Y, Rapoport SI, Smith QR. An in situ brain perfusion technique to study cerebrovascular transport in the rat. Am J Physiol. 1984;247(3 Pt 2):H484–93.PubMedGoogle Scholar
  30. 30.
    Smith QR, Allen DD. Methods in molecular medicine: the blood-brain barrier: biology and research protocols. In: Sukriti Nag, editors. Totowa, NJ: Humana Press Inc.; 2003. p. 209–218.Google Scholar
  31. 31.
    Krupka RM. Expression of substrate specificity in facilitated transport systems. JMB. 1990;117:69–78.Google Scholar
  32. 32.
    Liu X, Smith BJ, Chen C, Callegari E, Becker SL, Chen X, et al. Use of a physiologically based pharmacokinetic model to study the time to reach brain equilibrium: an experimental analysis of the role of blood-brain barrier permeability, plasma protein binding, and brain tissue binding. J Pharmacol Exp Ther. 2005;313(3):1254–62.PubMedCrossRefGoogle Scholar
  33. 33.
    Walker I, Nicholls D, Irwin WJ, Freeman S. Drug delivery via active transport at the blood-brain barrier: affinity of a prodrug of phosphonoformate for the large amino acid transporter. Int J Pharm. 1994;104(2):157–67.CrossRefGoogle Scholar
  34. 34.
    Ruocco LA, Viggiano D, Viggiano A, Abignente E, Rimoli MG, Melisi D, et al. Galactosylated dopamine enters into the brain, blocks the mesocorticolimbic system and modulates activity and scanning time in Naples high excitability rats. Neuroscience. 2008;152(1):234–44.PubMedCrossRefGoogle Scholar
  35. 35.
    Dalpiaz A, Pavan B, Scaglianti M, Vitali F, Bortolotti F, Biondi C, et al. Transporter-mediated effects of diclofenamic acid and its ascorbyl pro-drug in the in vivo neurotropic activity of ascorbyl nipecotic acid conjugate. J Pharm Sci. 2004;93(1):78–85.PubMedCrossRefGoogle Scholar
  36. 36.
    Manfredini S, Pavan B, Vertuani S, Scaglianti M, Compagnone D, Biondi C, et al. Design, synthesis and activity of ascorbic acid prodrugs of nipecotic, kynurenic and diclophenamic acids, liable to increase neurotropic activity. J Med Chem. 2002;45(3):559–62.PubMedCrossRefGoogle Scholar
  37. 37.
    van de Waterbeemd H, Smith DA, Jones BC. Lipophilicity in PK design: methyl, ethyl, futile. J Comput Aided Mol Des. 2001;15(3):273–86.PubMedCrossRefGoogle Scholar
  38. 38.
    Levin VA. Relationship of octanol/water partition coefficient and molecular weight to rat brain capillary permeability. J Med Chem. 1980;23(6):682–4.PubMedCrossRefGoogle Scholar
  39. 39.
    Elia J, Easley C, Kirkpatrick P. Lisdexamfetamine dimesylate. Nat Rev Drug Discov. 2007;6(5):343–4.PubMedCrossRefGoogle Scholar
  40. 40.
    Gong T, Huang Y, Zang Z, L L. Synthesis and characterization of 9-[P-(N, N-dipropylsulfamide)] benzoylamino-1,2,3,4–4H-acridine—a potential prodrug for the CNS delivery of tacrine. J Drug Target. 2004;12(3):177–82.PubMedCrossRefGoogle Scholar
  41. 41.
    Thurlow RJ, Hill DR, Woodruff GN. Comparison of the uptake of [ 3H]-gabapentin with the uptake of L-[ 3H]-leucine into rat brain synaptosomes. Br J Pharmacol. 1996;118(3):449–56.PubMedCrossRefGoogle Scholar
  42. 42.
    Su T, Lunney E, Campbell G, Oxender DL. Transport of gabapentin, a γ-amino acid drug, by system L α-amino acid transporters: a comparative study in astrocytes, synaptosomes, and CHO cells. J Neurochem. 1995;64(5):2125–31.PubMedCrossRefGoogle Scholar
  43. 43.
    Nutt JG, Fellman JH. Pharmacokinetics of levodopa. Clin Neuropharmacol. 1984;7(1):35–49.PubMedCrossRefGoogle Scholar
  44. 44.
    Gomes P, Soares-da-Silva P. Interaction between L-DOPA and 3-O-methyl-l-DOPA for transport in immortalised rat capillary cerebral endothelial cells. Neuropharmacology. 1999;38(9):1371–80.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Lauri Peura
    • 1
    Email author
  • Kalle Malmioja
    • 1
  • Kristiina Huttunen
    • 1
  • Jukka Leppänen
    • 1
  • Miia Hämäläinen
    • 1
  • Markus M. Forsberg
    • 1
  • Jarkko Rautio
    • 1
  • Krista Laine
    • 1
  1. 1.School of PharmacyUniversity of Eastern FinlandKuopioFinland

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