W146 (trifluoroacetate salt) is a selective antagonist of Sphingosine 1-phosphate receptors 1 (S1PR1) with an EC50 value of 398nM[1].W146 (trifluoroacetate salt) is widely used to investigate S1P/S1PR1-mediated biological processes.
In vitro, W146 (trifluoroacetate salt) (10μM) was employed for 30min in HEK293 and CHO-K1 cells stably expressing S1P receptor. W146 (trifluoroacetate salt) 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 (trifluoroacetate salt) (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 (trifluoroacetate salt) and siponimod significantly suppressed the production of interleukin-1β in activated microglia stimulated with lipopolysaccharide plus nigericin[3].W146 (trifluoroacetate salt) (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 (trifluoroacetate salt) abolished the protective role of S1P and plasma proteins on the glycocalyx[4].
In vivo, W146 (trifluoroacetate salt) (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 (trifluoroacetate salt) (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 (trifluoroacetate salt)-treated mice exhibited significantly exacerbated renal and hepatic injury[6]. W146 (trifluoroacetate salt) (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 (trifluoroacetate salt), 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 (trifluoroacetate salt), 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 (trifluoroacetate salt)), 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.
W146 (trifluoroacetate salt)是鞘氨醇-1-磷酸受体1（S1PR1）的选择性拮抗剂，EC50值为398nM[1]。W146 (trifluoroacetate salt)广泛用于研究S1P/S1PR1介导的生物学过程。
体外实验中，W146 (trifluoroacetate salt)（10μM）处理稳定表达S1P受体的HEK293和CHO-K1细胞30分钟，W146 (trifluoroacetate salt)完全阻断了水溶性、高选择性S1PR1激动剂CYM-5442或S1P诱导的S1PR1内化、磷酸化、泛素化以及p42/p44 MAPK的激活[2]。在siponimod处理前，W146 (trifluoroacetate salt)（1μM）预处理小胶质细胞30分钟，随后细胞单独用1μg/mL LPS刺激24小时，或1μg/mL LPS刺激3.5小时并在最后30分钟加入10μM炎症小体激活剂nigericin。W146 (trifluoroacetate salt)和siponimod的预处理显著抑制了经LPS加nigericin激活的小胶质细胞中白细胞介素-1β的产生[3]。W146 (trifluoroacetate salt)（10μM）处理培养的大鼠脂肪垫内皮细胞（RFPECs）和人冠状动脉内皮细胞10分钟，随后细胞暴露于S1P或含血浆蛋白的培养基中2小时，W146 (trifluoroacetate salt)消除了S1P和血浆蛋白对糖萼的保护作用[4]。
在体内实验中，W146 (trifluoroacetate salt)（10mg/kg）经腹腔注射给予非禁食的Swiss小鼠，诱导了显著但短暂的血液淋巴细胞减少，并伴随淋巴结中CD4+和CD8+淋巴细胞的平行增加[5]。W146 (trifluoroacetate salt)（0.1mg/kg）经腹腔注射给予C57BL/6小鼠，每日一次，连续3天，随后进行20分钟肾脏缺血及24小时再灌注处理。W146 (trifluoroacetate salt)处理的小鼠表现出明显加重的肾和肝损伤[6]。W146 (trifluoroacetate salt)（5mg/kg）在AMD3100给药前1小时经腹腔注射，显著增强了C57BL/6小鼠中Kit+/Sca-1+/Lin−（KSL）造血干细胞和祖细胞的动员，增幅约达8倍[7]。