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Amino Acids

, Volume 43, Issue 1, pp 207–218 | Cite as

Design, conformational studies and analysis of structure–function relationships of PTH (1–11) analogues: the essential role of Val in position 2

  • A. CaporaleEmail author
  • L. Gesiot
  • M. Sturlese
  • A. Wittelsberger
  • S. Mammi
  • E. Peggion
Original Article

Abstract

The N-terminal 1–34 segment of parathyroid hormone (PTH) is fully active in vitro and in vivo and it elicits all the biological responses characteristic of the native intact PTH. Recent studies reported potent helical analogues of the PTH (1–11) with helicity-enhancing substitutions. This work describes the synthesis, biological activity, and conformational studies of analogues obtained from the most active non-natural PTH (1–11) peptide H-Aib-Val-Aib-Glu-Ile-Gln-Leu-Nle-His-Gln-Har-NH2; specifically, the replacement of Val in position 2 with d-Val, l-(αMe)-Val and N-isopropyl-Gly was studied. The synthesized analogues were characterized functionally by in-cell assays and their structures were determined by CD and NMR spectroscopy. To clarify the relationship between the structure and activity, the structural data were used to generate a pharmacophoric model, obtained overlapping all the analogues. This model underlines the fundamental functional role of the side chain of Val2 and, at the same time, reveals that the introduction of conformationally constrained Cα-tetrasubstituted α-amino acids in the peptides increases their helical content, but does not necessarily ensure significant biological activity.

Keywords

PTH NMR analysis Pharmacophoric model αMeVal Conformational analogues Structure/activity analysis 

Abbreviations

Aib

α-Aminoisobutyric acid

CD

Circular dichroism

DIPEA

N,N-Diisopropylethylamine

DMF

N,N-Dimethylformamide

EDT

Ethanedithiol

Har

Homoarginine

HATU

1-[Bis(dimethylamino)methylene]-1H-7-azabenzotriazolium hexafluorophosphate 3-oxide

HBTU

1-[Bis(dimethylamino)methylene]-1H-benzotriazolium hexafluorophosphate 3-oxide

HOAt

1-Hydroxy-7-azabenzotriazole

HOBt

1-Hydroxybenzotriazole

Nle

Norleucine

NMP

N-Methyl-2-pyrrolidone

Nval

N-Isopropylglycine

PTH

Parathyroid hormone

TFA

Trifluoroacetic acid

TFE

2,2,2-trifluoroethanol

TIS

Triisopropylsilane

TMS

Trimethylsilane

Notes

Acknowledgments

The authors thank MIUR, Ministry of Education and University of Italy, for financial support, Dr. Barbara Biondi for her kind help in mass analyses and in the synthetic approach, Dr. Nereo Fiori for his initial NMR analyses and Prof. Stefano Moro and the Molecular Modelling section of the Department of Pharmaceutical Sciences (Padova, Italy) for computational support. In particular, the authors want to thank Prof. Claudio Toniolo for encouragement and helpful discussions. The authors declare that they have no conflict of interest.

Supplementary material

726_2011_1065_MOESM1_ESM.doc (2.4 mb)
Supplementary material 1 (DOC 2467 kb)

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • A. Caporale
    • 1
    Email author
  • L. Gesiot
    • 2
  • M. Sturlese
    • 2
  • A. Wittelsberger
    • 3
  • S. Mammi
    • 2
  • E. Peggion
    • 2
  1. 1.Department of Molecular Sciences and NanosystemsUniversità Ca’ Foscari di VeneziaVeniceItaly
  2. 2.Department of Chemical SciencesInstitute of Biomolecular Chemistry, CNR, University of PadovaPadovaItaly
  3. 3.Department of PhysiologyTufts University School of MedicineBostonUSA

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