W146

W146 is a selective antagonist of Sphingosine 1-phosphate receptors 1 (S1PR1) with an EC₅₀ value of 398nM^[1].W146 is widely used to investigate S1P/S1PR1-mediated biological processes.

In vitro, W146 (10μM) was employed for 30min in HEK293 and CHO-K1 cells stably expressing S1P receptor. W146 completely abolished a water-soluble highly-selective S1PR1 agonist CYM-5442- or S1P-induced S1PR1 internalization, phosphorylation and ubiquitination, and p42/p44 MAPK activation^[2]. W146 (1µM) pretreated microglia for 30min before siponimod treatment and then the cells were stimulated with 1µg/mL LPS alone (for 24h) or 1µg/mL LPS (for 3.5h) plus 10µM inflammasome activator nigericin (for the final 30min). Pretreatment with W146 and siponimod significantly suppressed the production of interleukin-1β in activated microglia stimulated with lipopolysaccharide plus nigericin^[3].W146 (10μM) was applied to cultured rat fat-pad endothelial cells (RFPECs) and human coronary artery endothelial cells for 10min and then the cells were exposed to S1P or plasma protein-containing media for 2 hours. W146 abolished the protective role of S1P and plasma proteins on the glycocalyx^[4].

In vivo, W146 (10mg/kg) was administered intraperitoneally into non fasted Swiss mice and induced a significant but transient blood lymphopenia in mice and a parallel increase in CD4⁺ and CD8⁺ lymphocytes in lymph nodes^[5].W146 (0.1mg/kg) was administrated into C57BL/6 mice via intraperitoneal injection daily for 3 days before animals subjected to 20 minutes of renal ischemia followed by 24 hours of reperfusion. W146-treated mice exhibited significantly exacerbated renal and hepatic injury^[6]. W146 (5mg/kg) injected intraperitoneally 1h before AMD3100 administration significantly enhanced the mobilization of Kit⁺/Sca-1⁺/Lin⁻ (KSL) hematopoietic stem and progenitor cells in C57BL/6 mice by approximately 8-fold^[7].

References:
[1] M Germana Sanna, et al. Enhancement of capillary leakage and restoration of lymphocyte egress by a chiral S1P1 antagonist in vivo. Nat Chem Biol. 2006 Aug;2(8):434-41. Epub 2006 Jul 9.
[2] Gonzalez-Cabrera P J, Jo E J, Sanna M G, et al. Full pharmacological efficacy of a novel S1P1 agonist that does not require S1P-like headgroup interactions. Mol Pharmacol. 2008 Nov;74(5):1308-18.
[3] Tarrasón G, Aulí M, Mustafa S, et al.The sphingosine-1-phosphate receptor-1 antagonist, W146, causes early and short-lasting peripheral blood lymphopenia in mice. Int Immunopharmacol. 2011 Nov;11(11):1773-9.
[4] Zeng Y, Adamson R H, Curry F R E, Tarbell J M.Sphingosine-1-phosphate protects endothelial glycocalyx by inhibiting syndecan-1 shedding. Am J Physiol Heart Circ Physiol. 2014 Feb;306(3):H363-72.
[5] Tarrasón G, Aulí M, Mustafa S,et al. The sphingosine-1-phosphate receptor-1 antagonist, W146, causes early and short-lasting peripheral blood lymphopenia in mice. Int Immunopharmacol. 2011 Nov;11(11):1773-9.
[6] Ham A, Kim M, Kim J Y, et al. Selective deletion of the endothelial sphingosine-1-phosphate 1 receptor exacerbates kidney ischemia-reperfusion injury. Kidney Int. 2014 Apr;85(4):807-23.
[7] Liu J J, Zhao J W, Lee J F, et al. 3-amino-4-(3-hexylphenylamino)-4-oxobutyl phosphonic acid (W146), a Selective Antagonist of Sphingosine-1-phospahte Receptor Subtype 1, Enhances AMD3100-stimulated Mobilization of Hematopoietic Stem Progenitor Cells in Animals. J Biochem Pharmacol Res. 2013 Dec;1(4):197-203.