Borst, R

Chen, W

Chizari, MM

Erpenbeck, VJ

Kazani, S

Luo, J

Sandham, D

Xue, L

Xue, Luzheng

Chen, Wentao

Luo, Jian

Chizari, Mohammad Mostafa

Oxford University Research Archive

Fevipiprant, a Selective Prostaglandin D2 Receptor 2 Antagonist, Potently Inhibits Chemotaxis and Cytokine Production by Tc2 cells (type-2 CD8+ lymphocytes)  Luzheng Xuea  Wentao Chena, Jian Luoa, Mohammad Mostafa Chizaria, Rowie Borsta, Shamsah Kazanib, Veit J Erpenbeckc and David A Sandhamb aRespiratory Medicine Unit, University of Oxford, UK; bNovartis Institutes for Biomedical Research, Cambridge MA, USA; cNovartis Pharma AG, Basel, Switzerland Rationale Tc2 cells are type-2 cytokine-secreting CD8+ T lymphocytes that highly express the prostaglandin D2 (PGD2) receptor 2 (DP2). Tc2s are enriched in peripheral blood and airways specifically in patients with severe asthma and persistent corticosteroid-insensitive eosinophilia in two independent cohorts.  Activation with PGD2 leads to cell migration and pro-inflammatory protein production, which promotes eosinophil recruitment and survival directly or indirectly, suggesting an important role for Tc2 cells in the pathogenesis of eosinophilic asthma and potential contribution to airway eosinophilia. Fevipiprant is a potent selective DP2 receptor antagonist that reduces airway inflammation in patients with persistent asthma and elevated sputum eosinophil counts. The drug is currently in Phase III clinical development for treatment of uncontrolled asthma. Here, we characterize the inhibition of Fevipiprant on DP2 pathway activated Tc2 cell migration and cytokine release, including ex vivo studies on samples from severe eosinophilic asthmatics. Methods CD3+CD8+CRTH2+ Tc2 cells were isolated from mononuclear cells of healthy volunteer blood and cultured. For chemotaxis assays, cell migration induced with PGD2 (100 nM) in the presence of Fevipiprant was determined using the IncuCyte live cell analysis system.  Protein and mRNA levels of cytokines IL-4, IL-5 and IL-13 after treatment with PGD2 (200 nM) in the presence of Fevipiprant for 4 h were measured with ELISA and qPCR respectively. Negative controls DP1 receptor agonist BW245C and DP1 antagonist BW868A confirmed the DP2 specificity of the effect; DP2 antagonist TM30089 was a positive control. For ex-vivo studies, patients meeting ATS/ERS severe asthma definition with sputum eosinophil count >3% were recruited. Peripheral blood was collected and treated with PGD2 (1 µM) in the presence or absence of Fevipiprant (1 µM) for 6 h followed by intracellular staining (ICS) for IL-5/13 for flow cytometry analysis. (Ethical approval: Leicestershire, Northamptonshire Research Ethics Committee 08/H0406/189) Results  Fevipiprant specifically inhibited DP2-mediated chemotaxis (Fig. 1A) and type-2 cytokine production (Fig. 1B) in cultured Tc2 with low nM potency, and showed significant inhibition of type-2 cytokine production ex vivo (Fig. 1C). Representative data is presented in the Figure. Conclusions Fevipiprant is a potent inhibitor of DP2 pathway mediated activation of Tc2 cells. The inhibitory potency on Tc2 migration is comparable with previous data on inhibition of PGD2 induced activation in human ILC2 and Th2 cells. Given the potential role of Tc2s in uncontrolled eosinophilic asthma, these data support further development of Fevipiprant and complement prior characterization with ILC2s, eosinophils and Th2 cells.   Figure 1. Fevipiprant inhibition of DP2-mediated Tc2 cell activation. A. Cultured Tc2 cell migration in response to 100 nM PGD2 is mediated by DP2 and was inhibited by Fevipiprant; B. IL-5 production in response to 200 nM PGD2 in cultured Tc2 cells was inhibited by Fevipiprant; C. IL-5/IL-13+CD8+ T cells detected in fresh PBMC in response to 1 µM PGD2 were reduced by Fevipiprant.  

English

Fevipiprant, a selective prostaglandin D(2)receptor 2 antagonist, potently inhibits chemotaxis and cytokine production by Tc2 cells (type-2 CD8(+)lymphocytes)

Fevipiprant, a selective prostaglandin D(2)receptor 2 antagonist, potently inhibits chemotaxis and cytokine production by Tc2 cells (type-2 CD8(+)lymphocytes)

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