S1PR1 ( EC50 = 13.8 nM )
SEW2871 selectively targets the S1P1 receptor (S1PR1) with high affinity (EC50 = 13.8 nM). It does not activate S1P2, S1P3, S1P4, or S1P5 receptors at concentrations up to 10 µM.

SEW2871 shows enhanced migration effects by upregulating smooth muscle α-actin, procollagen αI and αIII, and total collagen content in LX-2 cells, a human hepatic stellate cell line [2].
In vitro, SEW2871 activates the S1P1 receptor with an EC50 of 13.8 nM, leading to downstream signaling such as ERK activation. It is a cell-permeable compound. Its selectivity for S1P1 over other S1P receptors has been confirmed in binding assays.

SEW2871 (20 mg/kg, gavage, once daily for 2 weeks) improves established experimental ductitis in IL-10–/– mice [3]. SEW2871 (0.5 mg/kg IP once daily) SEW2871 (0-0.3 mg/kg IV for 2 weeks) inhibits amyloid beta (Aβ1-42) in a trajectory model of Alzheimer's disease Induced spatial impairment memory and hippocampal neuron loss [2]. ) attenuates LPS-induced acute stress-induced lung injury in C57Bl/6 electrodes by producing the required alveolar and vascular septal protection [2]. SEW2871 reduces CD4+ T cell respiration inside the electrode and effectively protects cardiovascular short circuit animal models: IL-10–/– (interleukin (IL)-10 gene deficiency) mice, mouse model of Crohn’s disease (CD) [3] Dosage: 20 mg/kg Administration method: gavage. , once daily for 2 weeks Results: Colitis ameliorated in IL-10–/– mice, associated with reduced serum amyloid A concentration, reduced colonic MPO concentration, peripheral CD4+CD45+ T cell depletion, and T cell homing to colon LP. Inhibits typical cytokine and p-STAT-3 expression of type 1 helper T (Th1) and Th17 cells, and significantly reduces TNF-α, IFN-γ, IL-1β, and IL-17A mRNA levels.
In vivo, SEW2871 reduces the number of lymphocytes in the blood, a characteristic effect of S1P1 agonism. It is used in research studies related to diabetes, Alzheimer's disease, liver fibrosis, and inflammation.

Receptor Binding Assay[2]
[33P]-S1P (final concentration of 83 pM) was added to serial dilutions of S1P or AFD(R) in a 96-well plate. Assays were initiated with the addition of 25 μg membrane protein of S1P1-CHO cell membranes in a final volume of 200 μl assay buffer (50 mM HEPES [pH 7.5], 5 mM MgCl2, 1 mM CaCl2, 15 mM sodium fluoride, and 1% fatty-acid-free bovine serum albumin). Binding mixtures were incubated for 60 min at room temperature and terminated by filtration over Packard GF/B filter plates as described.

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Rac-Activation Assay[2]
Cells were serum starved for 16 hr and stimulated with S1P (50 nM, 500 nM, or vehicle control) or SEW2871 (500 nM, 5 μM, or vehicle control) in the serum-free medium with 0.1% fatty-acid-free BSA. At 2 min and 5 min, cells were lysed in 500 μl ice-cold lysis buffer (50 mM Tris [pH 8.0], 500 mM NaCl, 0.5% NP-40, 1 mM EDTA, 1 mM MgCl2, and protease inhibitor cocktail) containing 20 μg GST-tagged p21 binding domain of p21-activated protein kinase 1 (PAK1) [47]. The cell lysates were centrifuged at 16,000 × g at 4°C for 3 min, and 40 μl prewashed 50% GSH-Sepharose beads were added to the supernatant. The mixture was incubated at 4°C for 30 min, and the beads were washed twice in lysis buffer. The proteins bound to the beads were eluted with SDS sample buffer. Activated Rac GTPases were detected by Western blotting with a mouse monoclonal anti-Rac antibody.

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Membrane Preparation and Receptor Activation Assay[2]
Membranes were prepared from CHO cells expressing human S1P1 for use in ligand and 35S-GTPγS binding studies as described. For 35S-GTPγS binding assay, serial dilutions of S1P, AFD(R), SEW2871, SEW2905, or SEW2898 were added to membranes (1 to 10 μg protein/well) and assayed as described.
In vitro enzyme/receptor binding assays for SEW2871 involve radioligand binding studies using membranes from cells expressing human or murine S1P1 receptors. The compound's ability to displace a specific radiolabeled S1P1 ligand (e.g., [³³P]S1P or [³H]S1P) is measured to determine binding affinity (Ki). Functional activity is assessed by GTPγS binding assays to measure receptor activation (EC50).

Western Blotting[2]
Control CHO cells and the CHO cells stably transfected with human S1P1 or S1P3 were cultured to 50% confluence on a 6-well plate in complete RPMI1640 supplemented with 10% FBS. Cells were serum starved for 16 hr and stimulated with S1P, AFD(R), or SEW2871 diluted to various concentrations in the serum-free medium with 0.1% fatty-acid-free BSA. At 5 min, 1 hr, or 4.5 hr, cells were lysed in 50 mM Tris (pH 8.0), 125 mM NaCl, 20 mM CHAPS, 2 mM dithiothreitol, 1 mM EDTA, 2 mM Na3VO4, 10 mM NaF, 1 mM PMSF, and protease inhibitor cocktail. For the study of pertussis toxin (PTx) inhibition, cells were incubated with PTx at the final concentration of 100 ng/ml for 3 hr, prior to agonist stimulation. Cell lysates were analyzed by Western blotting after separation on 10% SDS-PAGE with mouse monoclonal anti-phospho-ERK1/2 antibody and rabbit polyclonal anti-phospho-Akt antibody (BD Biosciences). Total ERK1 and ERK2 were detected with a rabbit-affinity-purified polyclonal anti-ERK antibody, and total Akt was detected with a rabbit-affinity-purified polyclonal anti-Akt1 antibody. Band intensities corresponding to pERK1, pERK2, and pAkt were quantitated by image analysis. Amounts of pERK1/2 and pAkt were normalized for the total amounts of ERK1/2 and Akt.

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Receptor Internalization and Recycling[2]
HEK293 cells that were stably transfected with GFP-tagged S1P1 were grown in DMEM medium with 2% charcoal-stripped serum for 2 days [28]. Cells were pretreated with cycloheximide (15 μg/ml) for 30 min to block the synthesis of new S1P1 and stimulated with 100 nM S1P, FTY720-phosphate, 500 nM SEW2871, or SEW2898 for 30 min. Cells were washed and replenished with plain DMEM and cycloheximide and incubated for indicated time points. The fluorescence of the recycled receptor at each time point was quantified and expressed as percent control.
In vitro cellular assays for SEW2871 are performed using cell lines expressing the S1P1 receptor, such as CHO or HEK-293 cells. Cells are treated with the compound, and the activation of downstream signaling pathways, such as ERK phosphorylation or cAMP modulation, is measured by Western blot or ELISA. The compound's ability to induce receptor internalization is also studied using fluorescently tagged receptors.

IL-10–/– (interleukin (IL)-10 gene-deficient) mice, a murine model of Crohn's disease (CD)
20 mg/kg
Gavage, once daily for 2 weeks

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SEW2871 was dissolved in 100% dimethyl sulphoxide and diluted with 50% Tween 20 for use. We chose a dose of SEW2871 (20 mg/kg/day for 2 weeks) by gavage for the treatment, which was well tolerated, as reported previously by Lien et al. 26. In brief, the mice were divided into three groups: (i) treatment group: IL-10–/– mice received SEW2871 by gavage (20 mg/kg/day); (ii) control group: IL-10–/– mice that received equal volumes of distilled water alone; and (iii) WT group: WT mice also received equal volumes of distilled water. After administration, the mice were killed by excessive anaesthesia with pentobarbital sodium and their blood and colons were collected for experiments[3].
In vivo animal experiments with SEW2871 commonly use mouse models of autoimmune diseases (e.g., experimental autoimmune encephalomyelitis, EAE) or models of organ fibrosis. The compound is administered orally, and its effects on lymphocyte trafficking, disease severity, and tissue pathology are assessed. For example, in EAE models, the compound's ability to reduce clinical scores and inflammatory cell infiltration into the central nervous system is measured.

SEW2871 is orally active, indicating good bioavailability. Its PK properties have been characterized in preclinical species, showing systemic exposure following oral administration. It is designed to penetrate tissues and modulate S1P1 receptor activity in vivo.

Comprehensive toxicological data for SEW2871 are not detailed in publicly available sources. As a research compound, its safety profile has been evaluated in animal models for specific disease studies. No significant toxicity has been reported in the available literature for the doses used in these studies.

[1]. Sphingosine 1-Phosphate Receptor Modulators and Drug Discovery. Biomol Ther (Seoul). 2017 Jan 1;25(1):80-90.

[2]. S1P1-selective in vivo-active agonists from high-throughput screening: off-the-shelf chemical probes of receptor interactions, signaling, and fate. Chem Biol. 2005 Jun;12(6):703-15.

[3]. Oral treatment with SEW2871, a sphingosine-1-phosphate type 1 receptor agonist, ameliorates experimental colitis in interleukin-10 gene deficient mice. Clin Exp Immunol. 2014 Jul;177(1):94-101.

5-[4-phenyl-5-(trifluoromethyl)-2-thiophenyl]-3-[3-(trifluoromethyl)phenyl]-1,2,4-oxadiazole is an oxadiazole cyclic compound. SEW2871 is a selective sphingosine-1-phosphate type 1 receptor (S1P1) agonist and has been shown to effectively protect the kidneys from ischemia-reperfusion injury by reducing CD4+ T cell infiltration in mice. However, the effect of SEW2871 on colitis remains unclear. This study aimed to investigate the effect of SEW2871 on established colitis in interleukin (IL)-10 gene-deficient (IL-10-/-) mice (a mouse model of Crohn's disease (CD)). SEW2871 was administered to IL-10(-/-) mice by gavage at a dose of 20 mg/kg/day for 2 weeks. The severity of colitis, serum amyloid A, tissue myeloperoxidase (MPO) levels, T cell counts in blood and lamina propria (LP), and the production of pro-inflammatory cytokines were assessed. Furthermore, the expression of phosphorylated signal transducer and activator of transcription (STAT)-3 (p-STAT-3) in lymphocytes isolated from the lamina propria was evaluated. Two weeks of SEW2871 administration ameliorated established colitis in IL-10(-/-) mice with decreased serum amyloid A concentrations, decreased colonic MPO concentrations, reduced peripheral CD4(+)CD45(+) T cells, and decreased T cell homing to the lamina propria. In addition, SEW2871 treatment also inhibited the expression of typical cytokines of T helper cells type 1 (Th1) and Th17 cells, as well as p-STAT-3. SEW2871 treatment improved experimental colitis established in IL-10(-/-) mice, potentially providing a novel treatment option for Crohn's disease (CD) in humans. [1]

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The key role of the sphingosine-1-phosphate (S1P) receptor S1P(1) in regulating lymphocyte migration was demonstrated using the S1P(1) selective nanomolar agonist SEW2871. Despite lacking a charged head group, the tetraaromatic compound SEW2871 can bind to and activate S1P(1) through hydrophobic and ion-dipole interactions. Both S1P and SEW2871 activate the ERK, Akt, and Rac signaling pathways and induce S1P(1) internalization and recycling, unlike FTY720-phosphate-induced receptor degradation. S1P and SEW2871 stimulate lymphocyte migration through receptor recycling. Modeling and mutagenesis studies of S1P(1) showed that residues binding to the S1P head group are essential for S1P and SEW2871 to activate kinases. Therefore, SEW2871 reproduced the role of S1P in all detected signaling pathways and had overlapping interactions with key head group binding receptor residues, suggesting that it replaced salt bridge interactions with ion-dipole interactions. [2]

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Sphingosine-1-phosphate (S1P) was initially found to be an intracellular second messenger, but it was subsequently discovered unexpectedly that S1P also has the role of a first messenger, which ultimately led to the cloning of its G protein-coupled receptor S1P₁₋₅. Molecular identification of the S1P receptor opened new avenues for the pathophysiological study of this lipid mediator. In vitro cell and molecular studies and in vivo studies in genetically defective mice have elucidated the cellular signaling pathways and pathophysiological significance of the S1P receptor. Another unexpected discovery was that fingolimod (FTY720) can modulate the S1P receptor, which has accelerated drug development in this field. Fingolimod was approved in 2010, becoming the first oral active drug approved for the treatment of relapsing-remitting multiple sclerosis. Its application in other diseases is currently undergoing clinical trials. In addition, some more selective S1P receptor modulators with better pharmacokinetic properties and fewer side effects are under development. Some of these drugs are being tested in clinical trials for multiple sclerosis and other autoimmune and inflammatory diseases such as psoriasis, Crohn's disease, ulcerative colitis, polymyositis, dermatomyositis, liver failure, kidney failure, acute stroke and transplant rejection. The authors of this article will discuss the latest advances in the development of drugs targeting the S1P receptor and highlight their potential clinical applications. [3]
SEW2871 is a research tool widely used to study the biological functions of the S1P1 receptor and its role in immune cell trafficking, inflammation, and disease. It is not an approved drug. Its high selectivity for S1P1 makes it a valuable compound for distinguishing S1P1-mediated effects from those of other S1P receptors.

C20H10F6N2OS

440.3616

440.041

C, 54.55; H, 2.29; F, 25.89; N, 6.36; O, 3.63; S, 7.28

256414-75-2

256414-75-2

4077460

White to light yellow solid powder

1.4±0.1 g/cm3

490.3±55.0 °C at 760 mmHg

94.5-95.3ºC

250.3±31.5 °C

0.0±1.2 mmHg at 25°C

1.535

8.42

0

10

3

30

583

0

S1C(C2=NC(C3C([H])=C([H])C([H])=C(C(F)(F)F)C=3[H])=NO2)=C([H])C(C2C([H])=C([H])C([H])=C([H])C=2[H])=C1C(F)(F)F

OYMNPJXKQVTQTR-UHFFFAOYSA-N

InChI=1S/C20H10F6N2OS/c21-19(22,23)13-8-4-7-12(9-13)17-27-18(29-28-17)15-10-14(11-5-2-1-3-6-11)16(30-15)20(24,25)26/h1-10H

5-[4-phenyl-5-(trifluoromethyl)thiophen-2-yl]-3-[3-(trifluoromethyl)phenyl]-1,2,4-oxadiazole

SEW2871; SEW2871; 256414-75-2; SEW2871; SEW 2871; 5-[4-Phenyl-5-(trifluoromethyl)-2-thienyl]-3-[3-(trifluoromethyl)phenyl]-1,2,4-oxadiazole; SEW-2871; 5-(4-Phenyl-5-(trifluoromethyl)thiophen-2-yl)-3-(3-(trifluoromethyl)phenyl)-1,2,4-oxadiazole; 5-[4-phenyl-5-(trifluoromethyl)thiophen-2-yl]-3-[3-(trifluoromethyl)phenyl]-1,2,4-oxadiazole; MFCD00096600; SEW2871

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: ~25 mg/mL (~56.8 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.72 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (4.72 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)