Novel antagonists for proteinase-activated receptor 2: Inhibition of cellular and vascular responses in vitro and in vivo

T. Kanke, M. Kabeya, S. Kubo, S. Kondo, K. Yasuoka, J. Tagashira, H. Ishiwata, M. Saka, T. Furuyama, T. Nishiyama, T. Doi, Y. Hattori, A. Kawabata, M.R. Cunningham, R.J. Plevin

Research output: Contribution to journalArticle

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Abstract

Proteinase-activated receptor 2 (PAR(2)) is a G-protein coupled receptor associated with many pathophysiological functions. To date, the development of PAR(2) antagonists has been limited. Here, we identify a number of novel peptide-mimetic PAR(2) antagonists and demonstrate inhibitory effects on PAR(2)-mediated intracellular signalling pathways and vascular responses. The peptide-mimetic compound library based on the structures of PAR(2) agonist peptides were screened for inhibition of PAR(2)-induced calcium mobilisation in human keratinocytes. Representative compounds were further evaluated by radioligand binding and inhibition of NF kappa B transcriptional activity and IL-8 production. The vascular effects of the antagonists were assessed using in vitro and in vivo models. Two compounds, K-12940 and K-14585, significantly reduced SLIGKV-induced Ca2+ mobilisation in primary human keratinocytes. Both K-12940 and K-14585 exhibited competitive inhibition for the binding of a high-affinity radiolabelled PAR(2)-ligand, [H-3]-2-furoyl-LIGRL-NH2, to human PAR(2) with K-i values of 1.94 and 0.627 mu M respectively. NF kappa B reporter activity and IL-8 production were also significantly reduced. Furthermore, relaxation of rat-isolated aorta induced by SLIGRL-NH2 was inhibited competitively by K-14585. K-14585 also significantly lowered plasma extravasation in the dorsal skin of guinea pigs and reduced salivation in mice. K-12940 and K-14585 antagonized PAR(2) competitively, resulting in inhibition of PAR(2)-mediated signalling and physiological responses both in vitro and in vivo. These peptide-mimetic PAR(2) antagonists could be useful in evaluating PAR(2)-mediated biological events and might lead to a new generation of therapeutically useful antagonists.
LanguageEnglish
Pages361-371
Number of pages11
JournalBritish Journal of Pharmacology
Volume158
Issue number1
DOIs
Publication statusPublished - Sep 2009

Fingerprint

PAR-2 Receptor
Blood Vessels
Peptides
NF-kappa B
seryl-leucyl-isoleucyl--glycyl-lysyl-valine
Interleukin-8
Keratinocytes
In Vitro Techniques
Salivation
Competitive Binding
G-Protein-Coupled Receptors
Libraries
Aorta

Keywords

  • Proteinase-activated receptor 2 (PAR2
  • antagonist
  • Ca2+ mobilization
  • keratinocytes
  • radioligand-binding
  • thrombin receptor
  • smooth-muscle
  • human keratinocytes
  • nitric-oxide
  • inflammation
  • par-2
  • secretion
  • peptide
  • cells
  • mouse

Cite this

Kanke, T. ; Kabeya, M. ; Kubo, S. ; Kondo, S. ; Yasuoka, K. ; Tagashira, J. ; Ishiwata, H. ; Saka, M. ; Furuyama, T. ; Nishiyama, T. ; Doi, T. ; Hattori, Y. ; Kawabata, A. ; Cunningham, M.R. ; Plevin, R.J. / Novel antagonists for proteinase-activated receptor 2 : Inhibition of cellular and vascular responses in vitro and in vivo. In: British Journal of Pharmacology. 2009 ; Vol. 158, No. 1. pp. 361-371.
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author = "T. Kanke and M. Kabeya and S. Kubo and S. Kondo and K. Yasuoka and J. Tagashira and H. Ishiwata and M. Saka and T. Furuyama and T. Nishiyama and T. Doi and Y. Hattori and A. Kawabata and M.R. Cunningham and R.J. Plevin",
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Kanke, T, Kabeya, M, Kubo, S, Kondo, S, Yasuoka, K, Tagashira, J, Ishiwata, H, Saka, M, Furuyama, T, Nishiyama, T, Doi, T, Hattori, Y, Kawabata, A, Cunningham, MR & Plevin, RJ 2009, 'Novel antagonists for proteinase-activated receptor 2: Inhibition of cellular and vascular responses in vitro and in vivo' British Journal of Pharmacology, vol. 158, no. 1, pp. 361-371. https://doi.org/10.1111/j.1476-5381.2009.00342.x

Novel antagonists for proteinase-activated receptor 2 : Inhibition of cellular and vascular responses in vitro and in vivo. / Kanke, T.; Kabeya, M.; Kubo, S.; Kondo, S.; Yasuoka, K.; Tagashira, J.; Ishiwata, H.; Saka, M.; Furuyama, T.; Nishiyama, T.; Doi, T.; Hattori, Y.; Kawabata, A.; Cunningham, M.R.; Plevin, R.J.

In: British Journal of Pharmacology, Vol. 158, No. 1, 09.2009, p. 361-371.

Research output: Contribution to journalArticle

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T1 - Novel antagonists for proteinase-activated receptor 2

T2 - British Journal of Pharmacology

AU - Kanke, T.

AU - Kabeya, M.

AU - Kubo, S.

AU - Kondo, S.

AU - Yasuoka, K.

AU - Tagashira, J.

AU - Ishiwata, H.

AU - Saka, M.

AU - Furuyama, T.

AU - Nishiyama, T.

AU - Doi, T.

AU - Hattori, Y.

AU - Kawabata, A.

AU - Cunningham, M.R.

AU - Plevin, R.J.

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N2 - Proteinase-activated receptor 2 (PAR(2)) is a G-protein coupled receptor associated with many pathophysiological functions. To date, the development of PAR(2) antagonists has been limited. Here, we identify a number of novel peptide-mimetic PAR(2) antagonists and demonstrate inhibitory effects on PAR(2)-mediated intracellular signalling pathways and vascular responses. The peptide-mimetic compound library based on the structures of PAR(2) agonist peptides were screened for inhibition of PAR(2)-induced calcium mobilisation in human keratinocytes. Representative compounds were further evaluated by radioligand binding and inhibition of NF kappa B transcriptional activity and IL-8 production. The vascular effects of the antagonists were assessed using in vitro and in vivo models. Two compounds, K-12940 and K-14585, significantly reduced SLIGKV-induced Ca2+ mobilisation in primary human keratinocytes. Both K-12940 and K-14585 exhibited competitive inhibition for the binding of a high-affinity radiolabelled PAR(2)-ligand, [H-3]-2-furoyl-LIGRL-NH2, to human PAR(2) with K-i values of 1.94 and 0.627 mu M respectively. NF kappa B reporter activity and IL-8 production were also significantly reduced. Furthermore, relaxation of rat-isolated aorta induced by SLIGRL-NH2 was inhibited competitively by K-14585. K-14585 also significantly lowered plasma extravasation in the dorsal skin of guinea pigs and reduced salivation in mice. K-12940 and K-14585 antagonized PAR(2) competitively, resulting in inhibition of PAR(2)-mediated signalling and physiological responses both in vitro and in vivo. These peptide-mimetic PAR(2) antagonists could be useful in evaluating PAR(2)-mediated biological events and might lead to a new generation of therapeutically useful antagonists.

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KW - Proteinase-activated receptor 2 (PAR2

KW - antagonist

KW - Ca2+ mobilization

KW - keratinocytes

KW - radioligand-binding

KW - thrombin receptor

KW - smooth-muscle

KW - human keratinocytes

KW - nitric-oxide

KW - inflammation

KW - par-2

KW - secretion

KW - peptide

KW - cells

KW - mouse

U2 - 10.1111/j.1476-5381.2009.00342.x

DO - 10.1111/j.1476-5381.2009.00342.x

M3 - Article

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EP - 371

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