Skip to main content
SpringerLink
Log in

Prodrug Design for Brain Delivery of Small- and Medium-Sized Neuropeptides

  • Protocol
  • First Online:
Neuropeptides

Part of the book series: Methods in Molecular Biology ((MIMB,volume 789))

Abstract

The blood–brain barrier (BBB) represents multiple barriers for drug delivery from the circulation. Peptides potentially useful to treat maladies of the brain are especially limited in their ability to cross the BBB due to several shortcomings. Specific delivery strategies have been conceived to outwit the BBB to target neuropeptides into the brain. It should be noted, however, that no unified method is possible for true brain-targeting of these fascinating biomolecules due to their structural features, properties, and intricate interplays among factors governing their entrance into and retention within the brain. In most brain-targeting prodrug approaches, a lipophilic and bioreversible moiety(ies) is covalently attached to the peptide that results in the complete loss of the innate biological activity of the parent peptide (prodrugs are inactive per definition) but significantly improves brain uptake and metabolic stability in the plasma and the interstitial fluid. Once the peptide prodrug has crossed the BBB, specific enzymes liberate the parent agent from its prodrug in the brain. To illustrate the applicability of the prodrug strategy for brain delivery of small neuropeptides, pGlu-Glu-Pro-NH2, [Glu2TRH], a thyrotropin-releasing hormone (TRH) analogue with a vast array of central activities, was chosen as an example. An ester prodrug provided significantly improved brain delivery compared to the unmodified parent peptide. The synthesis, in vitro and in vivo evaluations of this prodrug as specific examples are given for typical exploratory prodrug validation.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Strand, F.L. (2003) Neuropeptides: general characteristics and neuropharmaceutical potential in treating CNS disorders In: Prokai, L., and Prokai-Tatrai, K. (eds) Peptide transport and delivery into the central nervous system, Vol. 61, Birkhäuser, Basel.

    Google Scholar 

  2. Prokai, L. (2002) Targeting drugs into the central nervous system. In Muzykantow, V.R., and Torchilin, V. (eds) Biomedical aspects of drug targeting, Kluwer Academic Publishers, Dordrecht.

    Google Scholar 

  3. Kusuhara, H., and Sugiyama, Y. (2001) Efflux transport systems for drugs at the blood–brain barrier and blood–cerebrospinal fluid barrier (Part 1). Drug Discov. Today 6, 150–156.

    Google Scholar 

  4. Begley, D.J., and Brightman, M.W. (2003) Structural and functional aspects of the blood–brain barrier. In: Prokai, L., and Prokai-Tatrai, K. (eds) Peptide transport and delivery into the central nervous system, Vol. 61, Birkhäuser, Basel.

    Google Scholar 

  5. Brownlees, J., and Williams, C.H. (1993) Peptidases, peptides at the mammalian blood–brain barrier. J. Neurochem. 60, 793–803.

    Google Scholar 

  6. Pardridge, W.J. (2002) Drug and gene targeting to the brain with molecular Trojan horses. Nature Rev. Drug Discovery 1, 131–139.

    Google Scholar 

  7. Albert, A, (1958) Chemical aspects of selective toxicity. Nature 182, 421–422.

    Google Scholar 

  8. Prokai-Tatrai, K., and Prokai, L. (2003) Modifying peptide properties by prodrug design for enhanced transport into the CNS. In: Prokai, L., and Prokai-Tatrai, K. (eds) Peptide transport and delivery into the central nervous system, Vol. 61, Birkhäuser, Basel.

    Google Scholar 

  9. Wong, A., and Toth, I. (2001) Lipid, sugar and liposaccharide based delivery systems. Curr. Med. Chem. 8, 1123–1136.

    Google Scholar 

  10. Powell, M.F. (1993). Peptide stability in drug development: in vitro peptide degradation in plasma and serum. Ann. Rep. Med. Chem. 28, 285–294.

    Google Scholar 

  11. Prokai-Tatrai, K., Teixido, M., Nguyen, V. et al (2005) A pyridinium-substituted analog of the TRH-like tripeptide pGlu-Glu-Pro-NH2 and its prodrugs as central nervous system agents. Med. Chem. 1, 141–152.

    Google Scholar 

  12. Prokai-Tatrai, K., Nguyen, V., Zharikova, A.D. et al (2003) Prodrugs to enhance central nervous system effects of the TRH-like peptide pGlu-Glu-Pro-NH2. Bioorg. Med. Chem. Lett. 13, 1011–10144.

    Google Scholar 

  13. Prokai-Tatrai, K., and Prokai, L. (1996) Brain-targeted delivery of a leucine-enkephalin analogue by retrometabolic design. J. Med. Chem. 39, 4775–4782.

    Google Scholar 

  14. Prokai-Tatrai, K., Kim, H.S., and Prokai, L. (2008) The utility of oligopeptidase in brain-targeting delivery of an enkephalin analogue by prodrug design. Open Med. Chem. J. 2, 97–100.

    Google Scholar 

  15. Prokai, L., Prokai-Tatrai, K., Zharikova, A.D. et al. (2004) Centrally acting and metabolically stable thyrotropin-releasing hormone analogues by replacement of histidine with substituted pyridinium. J. Med. Chem. 47, 625–6033.

    Google Scholar 

  16. Klon, A.E. (2009) Computational models for central nervous system penetration. Curr. Med. Chem. 5, 71–89.

    Google Scholar 

  17. Danielsson, L.G., and Zhang, Y.H. (1996) Methods for determining n-octanol-water partition constants. TRAC–Trends Anal. Chem. 15, 188–196.

    Google Scholar 

  18. Ong, S., Hanlan, C., and Pidgeon, J. (1996) Immobilized-artificial-membrane chromatography: Measurements of membrane partition coefficient and predicting drug membrane permeability. J. Chromatogr. 728, 13–128.

    Google Scholar 

  19. Faller, B. (2008) Artificial membrane assays to assess permeability. Curr. Drug Metab. 9, 886–892.

    Google Scholar 

  20. Deli, M.A., Abraham, C.S., Kataoka, Y. et al. (2005) Permeability studies on in vitro blood-brain barrier models: Physiology, pathology, and pharmacology. Cell Mol. Neurobiol. 25, 59–129.

    Google Scholar 

  21. Bradbury, M.W.B., Patlak, C.S., and Oldendorf, W.H. (1975) Analysis of brain uptake and loss of radiotracers after intracarotid injection. Am. J. Physiol. 229, 1110–1115.

    Google Scholar 

  22. Prokai, L., Zharikova, A.D., Janaky T et al. (2000) Exploratory pharmacokinetics and brain distribution study of a neuropeptide FF antagonist by liquid chromatography/atmospheric pressure ionization tandem mass spectrometry. Rapid Commun. Mass Spectrom. 14, 2412–2418.

    Google Scholar 

  23. Hansch, C., and Clayton, J.M. (1973) Lipophilic character and biological-activity of drugs. 2. Parabolic case. J. Pharm. Sci. 62, 1–21.

    Google Scholar 

  24. Summerfield, S.G., Read, K., Begley, D.J. et al.(2007) Central nervous system drug disposition: The relationship between in situ brain permeability and brain free fraction. J. Pharmacol. Exp. Ther. 322, 205–213.

    Google Scholar 

  25. Del Rio-Garcia, J., and Smyth, D.G. (1990) Distribution of pyroglutamylpeptide amides related to thyrotropin-releasing hormone in the central nervous system and periphery of the rat. J. Endocrinol. 127, 445–450.

    Google Scholar 

  26. Prokai, L. (2002) Central nervous system effects of thyrotropin-releasing hormone and its analogues: opportunities and perspectives for drug discovery and development. In: Jucker, E.M. (ed) Progress in drug research, Vol 59, Birkhäuser, Basel.

    Google Scholar 

  27. Prokai-Tatrai, K., Prokai, L. (2009) Prodrugs of thyrotropin-releasing hormone and related peptides as central nervous system agents. Molecules 14, 633–654.

    Google Scholar 

  28. Kelly, J.A., Slator, G.R., Tipton, K.F. et al. (2000) Kinetic investigation of the specificity of porcine brain thyrotropin-releasing hormone-degrading ectoenzyme for thyrotropin-releasing hormone-like peptides. J. Biol. Chem. 275, 16746–16751.

    Google Scholar 

  29. Nguyen, V., Zharikova, A.D., and Prokai, L. (2007) Evidence for interplay between pGlu-Glu-Pro-NH2 and thyrotropin-releasing hormone in the brain. Neurosci. Lett. 415, 64–67.

    Google Scholar 

  30. Nguyen, V., Zharikova, A.D., Prokai-Tatrai, K. et al. (2010) [Glu2]TRH dose-dependently attenuates TRH-evoked analeptic effect in the mouse brain. Brain Res. Bull. 82, 83–86.

    Google Scholar 

  31. Merrifield, R.B. (1963) Solid phase peptide synthesis. Synthesis of a tetrapeptide. J. Am. Chem. Soc. 85, 2149–2154.

    Google Scholar 

  32. Carpino, L.A., and Han, G.Y. (1972) 9-Fluorenylmethoxycarbonyl amino-protecting group. J. Org. Chem. 37, 3404–3409.

    Google Scholar 

  33. Albericio, F., Bofill, J.M., El-Faham, A. et al. (1998) Use of onium salt-based coupling reagents in peptide synthesis. J. Org. Chem. 63, 9678–9683.

    Google Scholar 

  34. Kaiser, E., Colescott, R.L., Bossinger, C.D. et al. (1970) Color test for detection of free terminal amino groups in the solid-phase synthesis of peptides. Anal. Biochem. 34, 595–598.

    Google Scholar 

  35. Mant, C.T.; and Hodges, R.S. (1991). High-performance liquid chromatography of peptides and proteins; CRC Press: Boca Raton, Fl.

    Google Scholar 

  36. Prokai, L., Zharikova, A., Janáky, T. et al. (2001) Integration of mass spectrometry into early-phase discovery and development of central nervous system agents. J. Mass Spectrom. 36, 1211–1219.

    Google Scholar 

  37. Braddy, A.C., Janaky, T., and Prokai, L. (2002) Immobilized artificial membrane chromatography coupled with atmospheric pressure ionization mass spectrometry. J. Chromatogr. A 966, 81–87.

    Google Scholar 

  38. http://www.registech.com/InfoPages/IamInfo/IamCareAndUse.html

  39. Vojkovsky, T. (1995) Detection of secondary amine on solid phase. Pept. Res. 8, 236–237.

    Google Scholar 

  40. Cheng, Y., and Prusoff, W.H. (1973) Relationship between the inhibition constant (Ki) and the concentration of inhibitor which causes 50 percent inhibition (I50) of an enzymatic reaction. Biochem. Pharmacol. 22, 3099–3108.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pharmacology and Neuroscience, University of North Texas Health Science Center, Fort Worth, TX, USA

    Katalin Prokai-Tatrai & Laszlo Prokai

Authors
  1. Katalin Prokai-Tatrai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Laszlo Prokai
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Laszlo Prokai .

Editor information

Editors and Affiliations

  1. , Dipt. di Morfofisiologia Veterinaria, Università degli Studi di Torino, Via Leonardo da Vinci 44, Grugliasco, 10095, Torino, Italy

    Adalberto Merighi

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Prokai-Tatrai, K., Prokai, L. (2011). Prodrug Design for Brain Delivery of Small- and Medium-Sized Neuropeptides. In: Merighi, A. (eds) Neuropeptides. Methods in Molecular Biology, vol 789. Humana Press. https://doi.org/10.1007/978-1-61779-310-3_21

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-310-3_21

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-309-7

  • Online ISBN: 978-1-61779-310-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation