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Structural elucidation of Leuprolide and its analogues in solution: insight into their bioactive conformation

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Abstract

Leuprolide [dLeu6, NHEt10]GnRH, a potent gonadotropin-releasing hormone (GnRH) agonist, is used in a wide variety of hormone-related diseases like cancer and endometriosis. In this report, the conformational behaviour of Leuprolide and its linear synthetic analogues, namely [Tyr5(OMe), dLeu6, Aze9, NHEt10]GnRH (1) and [Tyr5(OMe), dLeu6, NHEt10]GnRH (2) have been studied in DMSO and H2O solutions by means of 2D nuclear magnetic resonance (NMR) experiments and detailed molecular dynamics (MD) simulations. The aim was to identify the conformational requirements of GnRH analogues for agonistic activity. This approach is of value as no crystallographic data are available for the GnRH receptor (G protein-coupled receptor, GPCR). The NOE data indicate the existence of a β-turn type I in the 2–5 segments of Leuprolide and its linear analogues in the case of using DMSO-d6 as solvent, whereas a β-turn type II in the 3–6 segments is indicated using D2O as solvent. The final structures fulfil the conformational requirements that are known, in the literature, to play a significant role in receptor recognition and activation. Finally, the linear analogues (1) and (2) are biologically active when tested against the human breast cancer cell line, MCF-7.

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Abbreviations

GnRH:

Gonadotropin releasing hormone

NMR:

Nuclear magnetic resonance

MD:

Molecular dynamics

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Acknowledgments

Despina Laimou is thankful for being supported by University of Patras (Grand K. Karatheodoris).

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Authors and Affiliations

  1. Department of Chemistry, University of Patras, Patras, 26500, Greece

    Despina K. Laimou, Maria Katsara, Minos-Timotheos I. Matsoukas & Theodore V. Tselios

  2. Immunology and Vaccine Laboratory, Burnet Institute (Austin Campus), Studley Road, Heidelberg, Victoria, 3084, Australia

    Maria Katsara & Vasso Apostolopoulos

  3. Department of Biological Applications and Technologies, University of Ioannina, Ioannina, 45110, Greece

    Anastassios N. Troganis

Authors
  1. Despina K. Laimou
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  2. Maria Katsara
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  3. Minos-Timotheos I. Matsoukas
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  4. Vasso Apostolopoulos
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  5. Anastassios N. Troganis
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  6. Theodore V. Tselios
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Corresponding authors

Correspondence to Anastassios N. Troganis or Theodore V. Tselios.

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Laimou, D.K., Katsara, M., Matsoukas, MT.I. et al. Structural elucidation of Leuprolide and its analogues in solution: insight into their bioactive conformation. Amino Acids 39, 1147–1160 (2010). https://doi.org/10.1007/s00726-010-0549-8

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  • DOI: https://doi.org/10.1007/s00726-010-0549-8

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