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Etrasimod, a Novel Sphingosine‐1‐Phosphate Receptor (S1PR1) Modulator, Lacks Functional Activity at S1P2 Receptors
Author(s) -
Adams John W,
Unett David J.,
Chen Xiaohua,
Gaidarov Ibragim
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.671.15
Subject(s) - sphingosine 1 phosphate receptor , s1pr1 , receptor , internalization , sphingosine 1 phosphate , fingolimod , microbiology and biotechnology , g protein coupled receptor , sphingosine , biology , pharmacology , chemistry , biochemistry , cancer research , immunology , multiple sclerosis , vascular endothelial growth factor a , vascular endothelial growth factor , vegf receptors
The S1P1 receptor modulator FTY720 (Fingolimod), a prodrug that is phosphorylated in vivo, possesses potent immunomodulatory activities and has been approved for the treatment of multiple sclerosis. FTY720 phosphate also possesses functional activity at S1P3, 4, and 5 receptor subtypes but, until recently, was thought to lack activity at the S1P2 subtype. We sought to develop methods to reliably detect functional activity at S1PR2 and to investigate whether structurally diverse S1PR1 modulators, including Etrasimod (APD334) a novel S1P receptor modulator, possess activity at this receptor. FTY720 phosphate and a panel of S1P receptor modulators were evaluated for their abilities to interact with S1PR2 using a broad panel of functional assays. Treatment of an S1PR2 expressing cell line with FTY720 phosphate produced rapid and robust receptor internalization. As observed with the S1P1 receptor, internalization was sustained following compound wash‐out. A number of additional S1PR modulators were found to induce the same internalization, while Etrasimod was inactive. Unlike the natural ligand S1P, these modulators failed to recruit b‐arrestin to S1PR2. However, label free Dynamic Mass Redistribution assays (Corning®, Epic®) readily detected S1PR2 mediated agonist like functional responses. Pretreatment of S1PR2 expressing cells with these modulators significantly blunted subsequent responses of S1PR2 toward its natural ligand S1P and reversed S1P‐mediated inhibition of fibronectin‐induced migration in S1P2 receptor expressing HEK293 cells. These data demonstrate that, in addition to FTY720, a wide range of S1PR modulators can also modulate S1PR2 while Etrasimod lacked such activity. Dynamic Mass Redistribution and receptor internalization assays appear suitable for quickly assessing S1PR2 activity. While S1PR1 is a chemotactic receptor, our data support the prevailing view that S1PR2 is a chemorepellant receptor, through which S1P negatively regulates cell migration. S1PR2 also mediates the antiproliferative activity of S1P in normal and cancer cell lines. S1PR2 ablation has been shown to result in stimulation of tumor development and metastasis in various animal models of cancer. Our data suggest that some S1PR1 modulators may have a negative effect on signaling pathways involved in regulation of cancer cell growth and metastasis. Support or Funding Information Funding: Arena Pharmaceuticals, Inc.