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Mimetics of the disulfide bridge between the N- and C-terminal cysteines of the KLK3-stimulating peptide B-2

Amino Acids volume 39, pages 233–242 (2010)Cite this article

Abstract

Human prostate produces kallikrein-related peptidase 3 (KLK3, also known as prostate specific antigen), which is widely used as a prostate cancer marker. Proteolytically active KLK3 has been shown to inhibit angiogenesis and its expression decreases in poorly differentiated tumors. Thus, it may be possible to control prostate cancer growth with agents that stimulate the proteolytic activity of KLK3. We have earlier developed synthetic peptides, which bind specifically to KLK3 and promote its proteolytic activity. These peptides are cyclic, all containing a disulfide bridge between the N- and C-terminal cysteines. To increase the in vivo stability of the KLK3-stimulating peptide B-2, we made differently cyclized analogues by replacing both terminal cysteines and the disulfide bridge between them. A replacement consisting of γ-amino butyric acid and aspartic acid, where the amino group from the former was linked to the main chain carboxyl group of the latter, was found to be, at high concentrations, more active than the B-2 peptide. Furthermore, as compared to the parent peptide, this analog had an improved stability in plasma and against the enzymatic degradation by KLK3. In addition, the series of analogues also provided valuable information of the structure–activity relationships of the B-2 peptide.

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Abbreviations

Acm:

Acetamidomethyl

Ahx:

6-Aminohexanoic acid

All:

Allyl

Aox:

8-Aminooctanoic acid

BHP:

Benign hyperplasia

DCM:

Dichloromethane

DIEA:

N,N-diisopropylethylamine

DMF:

N,N′-dimethylformamide

EDT:

Ethanedithiol

ESI:

Electrospray ionization

Fmoc:

Fluorenylmethoxycarbonyl

GABA:

γ-Amino butyric acid

HATU:

2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate

HBTU:

2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate

HLPC:

High-performance liquid chromatography

HOAt:

1-Hydroxy-7-azabenzotriazole

HUVEC:

Umbilical-vein endothelial cells

KLK2:

Kallikrein-related peptidase 2

KLK3:

Kallikrein-related peptidase 3

MALDI:

Matrix assisted laser desorption ionization

Mtt:

4-Methyltrityl

ODmab:

4{N-[1-(4,4-dimethyl-2,6-dioxo-cyclohexylidene)-3-methylbutyl]-amino}benzyloxy

PBS:

Phosphate buffered saline

PSA:

Prostate specific antigen

SPPS:

Solid-phase peptide synthesis

tBuO:

t-Butoxy

TFA:

Trifluoroacetic acid

TIS:

Triisopropyl silane

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Acknowledgments

This work was supported by Finnish Funding Agency for Technology and Innovation (TEKES), University of Helsinki, Helsinki University Central Hospital, the Finnish Cancer Foundation, the Academy of Finland (Grant No. 126969), Juselius Foundation, and Finska Läkaresällskapet. We would also like to thank J Rytkönen and A. Uljas for technical assistance.

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Affiliations

  1. Laboratory of Chemistry, Department of Biosciences and Biocenter Kuopio, University of Kuopio, P.O. Box 1627, 70211, Kuopio, Finland

    Miikka Pakkala, Janne Weisell, Jouko Vepsäläinen & Ale Närvänen

  2. Department of Biotechnology and Molecular Medicine, University of Kuopio, P.O. Box 1627, 70211, Kuopio, Finland

    Ale Närvänen

  3. Department of Clinical Chemistry, Biomedicum, University of Helsinki and Helsinki University Central Hospital, 00014, Helsinki, Finland

    Can Hekim, Ulf-Håkan Stenman & Hannu Koistinen

  4. Division of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Helsinki, 00014, Helsinki, Finland

    Erik A. A. Wallen

Authors
  1. Miikka Pakkala
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  2. Janne Weisell
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  3. Can Hekim
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  4. Jouko Vepsäläinen
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  5. Erik A. A. Wallen
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  6. Ulf-Håkan Stenman
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  7. Hannu Koistinen
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  8. Ale Närvänen
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Correspondence to Ale Närvänen.

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Pakkala, M., Weisell, J., Hekim, C. et al. Mimetics of the disulfide bridge between the N- and C-terminal cysteines of the KLK3-stimulating peptide B-2. Amino Acids 39, 233–242 (2010). https://doi.org/10.1007/s00726-009-0433-6

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  • DOI: https://doi.org/10.1007/s00726-009-0433-6

Keywords

  • Synthetic peptide
  • Stability
  • Prostate cancer
  • Kallikrein-related peptidase 3
  • KLK3
  • Prostate specific antigen
  • PSA