Novel antagonists for proteinase‐activated receptor 2: inhibition of cellular and vascular responses <i>in vitro</i> and <i>in vivo</i>

Kanke, Toru; Kabeya, Mototsugu; Kubo, Satoko; Kondo, Shinichi; Yasuoka, Kasane; Tagashira, J; Ishiwata, Hiroyuki; Saka, Masako; Furuyama, Takafumi; Nishiyama, Taihei; Doi, Takeshi; Hattori, Yukio; Kawabata, Atsufumi; Cunningham, Margaret; Plevin, Robin

doi:10.1111/j.1476-5381.2009.00342.x

Cited by 47 publications

(69 citation statements)

References 42 publications

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“…The peptide antagonist K-14585 was shown to inhibit PAR2-dependent IL-8 production, NF-κB phosphorylation, and p38 signaling (19). However, the K-14585 compound has partial agonist activity (19,38), as also observed with the wildtype PAR2 pepducin P2pal-21 (20). In this regard, we discovered that wild-type PAR2 has constitutive activity, indicating that certain extracellular or intracellular PAR2 ligands might stablize the latent on state.…”

Section: Discussionmentioning

confidence: 63%

“…In this model, PAR2 −/− mice had severe defects in their neutrophil and macrophage phagocytic ef- ficiency, suggesting that PAR2 may also confer protective effects in the host response to bacterial pathogens (37). Two other groups have disclosed PAR2 antagonists on the basis of the tethered peptide ligand (4,38). A PAR2 small molecule inhibitor, ENMD-1068, was tested in a model of joint inflammation (4).…”

Section: Discussionmentioning

confidence: 99%

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Interdicting protease-activated receptor-2-driven inflammation with cell-penetrating pepducins

Sevigny

Zhang

Bohm

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

108

136

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Protease-activated receptor-2 (PAR2), a cell surface receptor for trypsin-like proteases, plays a key role in a number of acute and chronic inflammatory diseases of the joints, lungs, brain, gastrointestinal tract, and vascular systems. Despite considerable effort by the pharmaceutical industry, PAR2 has proven recalcitrant to targeting by small molecule inhibitors, which have been unable to effectively prevent the interaction of the protease-generated tethered ligand with the body of the receptor. Here, we report the development of first-in-class cell-penetrating lipopeptide "pepducin" antagonists of PAR2. The design of the third intracellular (i3) loop pepducins were based on a structural model of a PAR2 dimer and by mutating key pharmacophores in the receptor intracellular loops and analogous pepducins. Individual pharmacophores were identified, which controlled constitutive, agonist, and antagonist activities. This approach culminated in the identification of the P2pal-18S pepducin which completely suppressed trypsin and mast cell tryptase signaling through PAR2 in neutrophils and colon cancer cells. The PAR2 pepducin was highly efficacious in blocking PAR2-dependent inflammatory responses in mouse models. These effects were lost in PAR2-deficient and mast-cell-deficient mice, thereby validating the specificity of the pepducin in vivo. These data provide proof of concept that PAR2 pepducin antagonists may afford effective treatments of potentially debilitating inflammatory diseases and serve as a blueprint for developing highly potent and specific i3-loop-based pepducins for other G protein-coupled receptors (GPCRs).protease-activated receptor-1 | protease-activated receptor-4

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Section: Discussionmentioning

confidence: 63%

Section: Discussionmentioning

confidence: 99%

Interdicting protease-activated receptor-2-driven inflammation with cell-penetrating pepducins

Sevigny

Zhang

Bohm

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

108

136

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“…More important, the developments in PAR-specific antagonists 26,27 enabled us to apply the PAR-2 inhibitor, FSLLRY-amide, which reversed already established PH in the hypoxic mouse model. Both PAR-2 Ϫ/Ϫ and FSLLRY-amide-treated wild-type mice exhibited decreased RVSP, normalization of cardiac performance, and diminished muscularization of the small pulmonary arteries.…”

Section: Discussionmentioning

confidence: 99%

PAR-2 Inhibition Reverses Experimental Pulmonary Hypertension

Kwapiszewska

Markart

Dahal

et al. 2012

Circulation Research

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Rationale: A hallmark of the vascular remodeling process underlying pulmonary hypertension (PH) is the aberrant proliferation and migration of pulmonary arterial smooth muscle cells (PASMC). Accumulating evidence suggests that mast cell mediators play a role in the pathogenesis of PH.Objective: In the present study we investigated the importance of protease-activated receptor (PAR)-2 and its ligand mast cell tryptase in the development of PH. Methods and Results:Our results revealed strong increase in PAR-2 and tryptase expression in the lungs of idiopathic pulmonary arterial hypertension (IPAH) patients, hypoxia-exposed mice, and monocrotaline (MCT)-treated rats. Elevated tryptase levels were also detected in plasma samples from IPAH patients. Hypoxia and platelet-derived growth factor (PDGF)-BB upregulated PAR-2 expression in PASMC. This effect was reversed by HIF (hypoxia inducible factor)-1␣ depletion, PDGF-BB neutralizing antibody, or the PDGF-BB receptor antagonist Imatinib. Attenuation of PAR-2 expression was also observed in smooth muscle cells of pulmonary vessels of mice exposed to hypoxia and rats challenged with MCT in response to Imatinib treatment. Tryptase induced PASMC proliferation and migration as well as enhanced synthesis of fibronectin and matrix metalloproteinase-2 in a PAR-2-and ERK1/2-dependent manner, suggesting that PAR-2-dependent signaling contributes to vascular remodeling by various mechanisms. Furthermore, PAR-2 ؊/؊ mice were protected against hypoxia-induced PH, and PAR-2 antagonist application reversed established PH in the hypoxia mouse model. Key Words: pulmonary arterial hypertension Ⅲ protease-activated receptor-2 Ⅲ mast cells P ulmonary hypertension (PH) is characterized by a persistent increase in pulmonary arterial pressure, which can ultimately lead to right ventricular failure and death. Factors that contribute to the increased vascular resistance in PH include vasoconstriction, vascular remodeling, and thrombosis. 1 Pulmonary vasoconstriction can result from the imbalance of vasoactive mediators such as nitric oxide, endothelin-1, prostacyclin, and serotonin. Another potent stimulator of arterial contraction is alveolar hypoxia. Low oxygen levels down-regulate voltage-gated potassium channels, leading to depolarization of smooth muscle cells (SMC), subsequent Ca 2ϩ influx, and vasoconstriction. [2][3][4] Prolonged contraction stimulates the proliferation and migration of SMC and fibroblasts leading to medial or adventitial thickening of the pulmonary arteries. 1 Remodeling can also be triggered by a dysfunction of endothelial cells, thereby altering vascular homeostasis by changing cytokine/growth factor levels and procoagulant potential. 1 Vascular damage and endothelial dysfunction can alter the composition of the extracellular matrix by the induction of matrix metalloproteinases (MMPs), resulting in SMC migration and proliferation. 5 Accumulating evidence suggests that inflammatory cells may play an active role in the development of PH. 6,7 The number of mast cells...

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“…To definitively determine whether the protective effects of PAR 2 on AHR are independent of β-arrestins in vivo will likely require using a model that results in a more robust AHR phenotype in β-arrestin-2 −/− mice. PAR 2 is up-regulated in the airways of patients with chronic asthma (35), which along with the reported proinflammatory effects of PAR 2 , has led to an interest in inhibitors of PAR 2 -activating proteinases or of the receptor itself as therapeutic agents for treating asthma (36). In contrast, the protective effects of PAR 2 have also generated interest in PAR 2 agonists as therapeutic agents for asthma, with the idea being that agonists of PAR 2 might promote bronchodilatation (2,17,37,38).…”

Section: Cells) (B) Graphmentioning

confidence: 99%

β-Arrestin-2 mediates the proinflammatory effects of proteinase-activated receptor-2 in the airway

Nichols

Saffeddine

Theriot

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

105

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Proteinase-Activated rreceptor-2 (PAR 2 ), a G-protein–coupled Receptor, activated by serine proteinases, is reported to have both protective and proinflammatory effects in the airway. Given these opposing actions, both inhibitors and activators of PAR 2 have been proposed for treating asthma. PAR 2 can signal through two independent pathways: a β-arrestin–dependent one that promotes leukocyte migration, and a G-protein/Ca 2+ one that is required for prostaglandin E 2 (PGE 2 ) production and bronchiolar smooth muscle relaxation. We hypothesized that the proinflammatory responses to PAR 2 activation are mediated by β-arrestins, whereas the protective effects are not. Using a mouse ovalbumin model for PAR 2 -modulated airway inflammation, we observed decreased leukocyte recruitment, cytokine production, and mucin production in β-arrestin-2 −/− mice. In contrast, PAR 2 -mediated PGE 2 production, smooth muscle relaxation, and decreased baseline airway resistance (measures of putative PAR 2 “protective” effects) were independent of β-arrestin-2. Flow cytometry and cytospins reveal that lung eosinophil and CD4 T-cell infiltration, and production of IL-4, IL-6, IL-13, and TNFα, were enhanced in wild-type but not β-arrestin-2 −/− mice. Using the forced oscillation technique to measure airway resistance reveals that PAR 2 activation protects against airway hyperresponsiveness by an unknown mechanism, possibly involving smooth muscle relaxation. Our data suggest that the PAR 2 -enhanced inflammatory process is β-arrestin-2 dependent, whereas the protective anticonstrictor effect of bronchial epithelial PAR 2 may be β-arrestin independent.

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

Cited by 47 publications

References 42 publications

Interdicting protease-activated receptor-2-driven inflammation with cell-penetrating pepducins

Interdicting protease-activated receptor-2-driven inflammation with cell-penetrating pepducins

PAR-2 Inhibition Reverses Experimental Pulmonary Hypertension

β-Arrestin-2 mediates the proinflammatory effects of proteinase-activated receptor-2 in the airway

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