The Effect of Glycosylation on the Uptake of an Enkephalin Analogue into the Central Nervous System

  • Sarah A. Williams
  • Thomas J. Abbruscato
  • Lajos Szabo
  • Robin Polt
  • Victor Hruby
  • Thomas P. Davis
Part of the Advances in Behavioral Biology book series (ABBI, volume 46)


In contrast to unglycosylated controls, glycosylated [D-Cys2,5]enkephalin-ser-gly (glycosylated DCDCE-ser-gly) elicits analgesia after intraperitoneal administration. This was postulated to be due to the presence of the glucose moiety allowing the analogue to cross the BBB via the glucose carrier. To test this hypothesis, the present study investigated the biological stability and the CNS uptake ofunglycosylated and glycosylated DCDCE-ser-gly. Interestingly, the metabolic half-lives and ability to cross the in vitro BBB was found to be similar for both analogues. In situ brain perfusion indicated that the brain uptake of glycosylated DCDCE-ser-gly was greater than that for the vascular marker, [14C]sucrose, but similar to the CSF uptake of the peptide. CNS uptake of glycosylated DCDCE-ser-gly was not affected by replacing D- with L- glucose, nor with the addition of 10 µM unlabelled glycosylated DCDCE-ser-gly. In summary, the difference in analgesic response of glycosylated compared to unglycosylated DCDCE-ser-gly, is not related to either differing metabolic profiles, nor the ability of the glycosylated analogue to use the glucose carrier to enter the CNS. However, this study does not eliminate the involvement of a different low affinity, saturable uptake system taking the glycosylated, but not the unglycosylated form.


Permeability Coefficient Brain Perfusion Brain Uptake Perfusion Medium Brain Microvessel Endothelial Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Après administration par voie intrapéritonéale, le peptide glycosylé (D-cys 2,5)enképhaline-ser-gly (DCDCE-ser-gly) montre une activité analgésique, contrairement au même peptide non-glycosylé. L’effet pharmacologique spécifique de l’enképhaline glycosylée pourrait être expliqué par l’addition du radical glycosylé qui permettrait àl’enképhaline de traverser la barrière hémato-encéphalique (BHE) en utilisant le transporteur du glucose. Afin de vérifier cette hypothèse, la stabilité ainsi que la pénétration cérébrale des molécules glycosylées et non glycosylées ont été étudiées. La demi-vie métabolique et la perméabilité de la BHE pour ces deux composés est semblable. Des études de microdialyse dans le cerveau montrent que la capture cérébrale du dérivé glycosylé est superieure à celle du (14 C)sucrose, mais similaire àla capture du peptide par le liquide cérébro-spinal. De plus, la capture cérébrale de la molécule glycosylée marquée n'est pas inhibée par le remplacement du D-glucose par du L-glucose ni par la dilution de la molécule marquée avec 10mM d'enképhaline glycosylée non marquée. En conséquence, les différences de réponse analgésique observées entre la DCDCE glycosylée et non glycosylée ne sont pas liées à une plus grande stabilité de la molécule glycosylée, ni au transport spécifique de ce composé par le transporteur du glucose de la BHE. Cependant cette étude n'écarte pas la possibilité du transport specifique du dérivé glycosylé par un autre système de transport saturable à faible affinité.


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  1. 1.
    Bell G. I., Burant C. F., Takeda J. and Gould G. W. (1993) Structure and function of mammalian facilitative sugar transporters. J. Biol. Chem. 268, 19161 - 19164.PubMedGoogle Scholar
  2. 2.
    Dayson H., Kleeman C. R. and Levin E. (1961) Blood-brain barrier and extracellular space. J. Physiol. 159, 67P - 68 P.Google Scholar
  3. 3.
    Dooley C. T., Chung N. N., Wilkes B. C., Schiller P. W., Bidlack J. M., Pasternak G. W. and Houghten R. A. (1994) An all D-amino acid opioid peptide with central analgesic activity from a combinatorial library. Science. 266, 2019 - 2022.PubMedCrossRefGoogle Scholar
  4. 4.
    Gerhart D. Z., Broderius M. A., Borson N. D. and Drewes L. R. (1992) Neurons and microvessels express the brain glucose transporter protein GLUT-3. Proc. Natl. Acad. Sci. 89, 733 - 737.PubMedCrossRefGoogle Scholar
  5. 5.
    Harik S. I., Hall A. K. and Perry G. (1992) Ontogeny of glucose transporter in the rat central nervous system. Neurology. 42, 407.CrossRefGoogle Scholar
  6. 6.
    Maher F., Vannucci S. J. and Simpson I. A. (1994) Glucose transporter proteins in brain. FASEB J. 8, 1003 - 1011.PubMedGoogle Scholar
  7. 7.
    Mantych G. J., James D. E. and Devaskar S. U. (1993) Jejunal/kidney glucose transporter isoform (Glut-5) is expressed in the human blood-brain barrier. Endocrinology. 132, 35 - 40.PubMedCrossRefGoogle Scholar
  8. 8.
    Pardridge W. M., Triguero D., Yang J. and Cancilla P. A. (1990) Comparison of in vitro and in vivo models of drug transcytosis through the blood-brain barrier. J. Pharmacol. Exp. Ther. 253, 884 - 891.PubMedGoogle Scholar
  9. 9.
    Pelligrino D. A., LaManna J. C., Duckrow R. B., Bryan Jr. R. M. and Harik S. I. (1992) Hyperglycemia and blood-brain barrier glucose transport. J. Cerebr. Blood Flow and Metab. 12, 887 - 899.Google Scholar
  10. 10.
    Poduslo J. F. and Curran G. L. (1994) Glycation increases the permeability of proteins across the blood-nerve and blood-brain barriers. Mol. Brain Res. 23, 157 - 162.PubMedCrossRefGoogle Scholar
  11. 11.
    Polt, R., Porreca, F., Szabò, L. Z., Bilsky E. J., Davis, P., Abbruscato, T. J., Davis, T. P., Horvath, R., Yamamura, H.I. and Hruby, V.J. (1994). Glycopeptide enkephalin analogues produce analgesia in mice: Evidence for penetration of the blood-brain barrier. Proc. Natl. Acad. Sci. USA. 91, 7114 - 7118.PubMedCrossRefGoogle Scholar
  12. 12.
    Preston J. E., Al-Sarraf H. and Segal M. B. (1995) Permeability of the developing blood-brain barrier to 14C-mannitol using the rat in situ brain perfusion technique. Dev. Brain Res. 87, 69 - 76.CrossRefGoogle Scholar
  13. 13.
    Raub T. J., Kuentzcl S. L. and Sawada G. A. (1992) Permeability of bovine brain microvessel endothelial cells in vitro: barrier tightening by a factor released from astroglioma cells. Exp. Cell Res. 199, 330 - 340.PubMedCrossRefGoogle Scholar
  14. 14.
    Weber S. J., Abbruscato T. J., Brownson E. A., Lipkowski A. W., Polt R., Misicka A., Haaseth R. C., Bartosz H., Hruby V. J. and Davis T. P. (1993) Assessment of an in vitro blood-brain barrier model using several [Mets]enkephalin opioid analogs. J. Pharmacol. Exp. Ther 266, 1649 - 1655.PubMedGoogle Scholar
  15. 15.
    Weber S. J., Greene D. L., Hruby V. J., Yamamura H. I., Porreca F. and Davis T. P. (1992) Whole body and brain distribution of [3H]Cyclic[D-Pen2,D-Pen']enkephalin after intraperitoneal. intravenous, oral and subcutaneous administration. J. Pharmacol. Exp. Ther. 263, 1308 - 1316.PubMedGoogle Scholar
  16. 16.
    Weiler-Güttler H., Zinke H., Möckel B., Frey A. and Gassen H. G. (1989) cDNA cloning and sequence analysis of the glucose transporter from porcine blood-brain barrier. Biol. Chem. Hoppe-Sevier 370, 467 - 473.Google Scholar
  17. 17.
    Williams S. A., Abbruscato T. J., Hruby V. J. and Davis T. P. (1995) The passage of a delta-opioid receptor selective enkephalin, DPDPE, across the blood-brain and blood-cerebrospinal fluid barriers. J. Neurochem. (in press).Google Scholar
  18. 18.
    Zlokovic B. V., Begley D. J., Djuricic B. M. and Mitrovic D. M. (1986) Measurement of solute transport across the blood-brain barrier in the perfused guinea-pig brain: method and application to N-methylaminoisobutyric acid. J. Neurochem. 46, 1444 - 1451.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1996

Authors and Affiliations

  • Sarah A. Williams
    • 1
  • Thomas J. Abbruscato
    • 1
  • Lajos Szabo
    • 2
  • Robin Polt
    • 2
  • Victor Hruby
    • 2
  • Thomas P. Davis
    • 1
  1. 1.Department of PharmacologyUniversity of ArizonaTucsonUSA
  2. 2.Department of ChemistryUniversity of ArizonaTucsonUSA

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