title: Characterization of selective Calcium-Release Activated Calcium channel blockers in mast cells and T-cells from human, rat, mouse and guinea-pig preparations.
abstract: Loss of function mutations in the two key proteins which constitute Calcium-Release Activated Calcium (CRAC) channels demonstrate the critical role of this ion channel in immune cell function. The aim of this study was to demonstrate that inhibition of immune cell activation could be achieved with highly selective inhibitors of CRAC channels in vitro using cell preparations from human, rat, mouse and guinea-pig. Two selective small molecule blockers of CRAC channels; GSK-5498A and GSK-7975A were tested to demonstrate their ability to inhibit mediator release from mast cells, and pro-inflammatory cytokine release from T-cells in a variety of species. Both GSK-5498A and GSK-7975A completely inhibited calcium influx through CRAC channels. This led to inhibition of the release of mast cell mediators and T-cell cytokines from multiple human and rat preparations. Mast cells from guinea-pig and mouse preparations were not inhibited by GSK-5498A or GSK-7975A; however cytokine release was fully blocked from T-cells in a mouse preparation. GSK-5498A and GSK-7975A confirm the critical role of CRAC channels in human mast cell and T-cell function, and that inhibition can be achieved in vitro. The rat displays a similar pharmacology to human, promoting this species for future in vivo research with this series of molecules. Together these observations provide a critical forward step in the identification of CRAC blockers suitable for clinical development in the treatment of inflammatory disorders.
authors: Respiratory Therapy Area Unit, GlaxoSmithKline, Medicines Research Centre, Gunnels Wood Road, Stevenage SG1 2NY, UK.
authors: Louise V
authors: LV
authors: Rice
authors: Heather J
authors: HJ
authors: Bax
authors: Linda J
authors: LJ
authors: Russell
authors: Victoria J
authors: VJ
authors: Barrett
authors: Sarah E
authors: SE
authors: Walton
authors: Angela M
authors: AM
authors: Deakin
authors: Sally A
authors: SA
authors: Thomson
authors: Fiona
authors: F
authors: Lucas
authors: Roberto
authors: R
authors: Solari
authors: David
authors: D
authors: House
authors: Malcolm
authors: M
authors: Begg
mesh_terms: D000818:Animals; D001549:Benzamides / Q000494:pharmacology*; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D015220:Calcium Channels / Q000378:metabolism*; D002460:Cell Line; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D005260:Female; D006168:Guinea Pigs; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism; D008168:Lung / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008407:Mast Cells / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010047:Ovalbumin; D011720:Pyrazoles / Q000494:pharmacology*; D051381:Rats; D013154:Spleen / Q000166:cytology; D013601:T-Lymphocytes / Q000187:drug effects* / Q000378:metabolism; D014132:Trachea / Q000187:drug effects / Q000502:physiology
chemical_list: C583715:2,6-difluoro-N-(1-(2-fluoro-6-(trifluoromethyl)benzyl)-1H-pyrazol-3-yl)benzamide; C583716:2,6-difluoro-N-(1-(4-hydroxy-2-(trifluoromethyl)benzyl)-1H-pyrazol-3-yl)benzamide; D001549:Benzamides; D002121:Calcium Channel Blockers; D015220:Calcium Channels; D016207:Cytokines; D011720:Pyrazoles; D010047:Ovalbumin