| Targets |
S1P1[1]
S1P1 agonist 1 targets the sphingosine-1-phosphate receptor 1 (S1P1), a G protein-coupled receptor (GPCR) that plays a critical role in lymphocyte egress from lymph nodes and the thymus. Upon agonist binding, S1P1 is internalized and downregulated, leading to retention of lymphocytes in lymphoid organs and reduced circulating lymphocyte counts. This mechanism is the basis for the immunomodulatory effects of S1P1 agonists such as fingolimod (FTY720). |
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| ln Vitro |
S1P1 Agonist 1 binds to S1P1 is internalized and activates intracellular AKT and ERKs cellular signaling pathways. S1P1 Agonist 1 mediated S1P1 downregulation is independent of sphingosine kinase activity indicating it to be a direct agonist of S1P1. S1P1 Agonist 1 decreases cell surface expression of S1P1 in in vitro cell culture model[1].
S1P1 agonist 1 binds to S1P1, is internalized, and activates intracellular AKT and ERK cellular signaling pathways. It mediates S1P1 downregulation independent of sphingosine kinase activity, indicating it is a direct agonist of S1P1. It decreases cell surface expression of S1P1 in in vitro cell culture models. The compound induces S1P1 internalization with an EC50 of 9.83 nM, demonstrating potent agonist activity at the receptor. |
| ln Vivo |
S1P1 Agonist 1 (1.3mg/kg) attenuates EAE disease. EAE is the animal model of multiple sclerosis which is extensively used for investigating clinical studies for multiple sclerosis drugs. S1P1 Agonist 1 treatment reduces peripheral total blood lymphocyte and T lymphocyte counts significantly. The reduction in total lymphocytes and T cells are 48% and 41% in AKP-11 treated animals, respectively. The reduction in the CD4+ and CD8+ T cell populations are 41% and 40% in S1P1 Agonist 1 treated animals, respectively. S1P1 Agonist 1 treatment reduces CNS infiltration of T cells and Cytokines and enhances neuroprotection. S1P1 Agonist 1 treatment has little effect on the heart rate of animals. The decrease in blood pressure is also smaller with S1P1 Agonist 1 treatment as compared animals treated with FTY720[1].
S1P1 agonist 1 (1.3 mg/kg) attenuates EAE disease in the rat model of multiple sclerosis. Treatment with S1P1 agonist 1 significantly reduces peripheral total blood lymphocyte and T lymphocyte counts, with reductions of 48% and 41%, respectively, in treated animals. The CD4+ and CD8+ T cell populations are reduced by 41% and 40%, respectively. S1P1 agonist 1 treatment reduces CNS infiltration of T cells and cytokines, enhances neuroprotection, and has little effect on heart rate, with a smaller decrease in blood pressure compared to FTY720. |
| Enzyme Assay |
Standard S1P1 receptor internalization assay: CHO-K1 cells stably expressing human S1P1-GFP (green fluorescent protein fusion) are seeded in 96-well plates. The next day, cells are treated with varying concentrations of S1P1 agonist 1 (0.0001-1000 nM) at 37degC for 30-60 minutes. Cells are fixed with 4% paraformaldehyde, and S1P1-GFP fluorescence is quantified using a fluorescence plate reader or high-content imaging system. EC50 for receptor internalization is calculated. Alternatively, a [3⁵S]GTPgammaS binding assay can be used to measure receptor activation.
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| Cell Assay |
Standard S1P1 signaling assay: Cells expressing S1P1 (e.g., CHO-K1-S1P1 cells) are seeded in 96-well plates and serum-starved overnight. Cells are treated with varying concentrations of S1P1 agonist 1 (0.0001-1000 nM) for 5-15 minutes. Cells are lysed, and AKT phosphorylation (Ser473) and ERK1/2 phosphorylation (Thr202/Tyr204) are measured by ELISA or Western blot. EC50 for activation of downstream signaling pathways is calculated. For cAMP inhibition assays, cells are pre-treated with test compound followed by forskolin stimulation, and cAMP levels are measured by HTRF.
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| Animal Protocol |
Rat experimental autoimmune encephalomyelitis (EAE) model: Female Lewis rats (6-8 weeks old, 150-200 g) are immunized subcutaneously with guinea pig myelin basic protein (MBP) or myelin oligodendrocyte glycoprotein (MOG) emulsified in complete Freund's adjuvant (CFA). On day 0, pertussis toxin is injected intravenously. S1P1 agonist 1 is administered orally at 1.3 mg/kg once daily starting from day 0 or day 7 post-immunization. Clinical signs of EAE (tail weakness, hind limb paralysis, incontinence) are scored daily (0-5 scale) for 18 days. At the end of the study, spinal cord sections are analyzed for inflammatory cell infiltration by H&E staining and for demyelination by Luxol fast blue staining.
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| ADME/Pharmacokinetics |
No specific PK data was found. S1P1 agonist 1 is described as orally active, indicating good oral bioavailability. FTY720, a related S1P receptor agonist, has an oral bioavailability of approximately 40% in rats and a half-life of ~30 hours in humans after phosphorylation. S1P1 agonist 1 is expected to have a similar PK profile: well-absorbed, extensively distributed, metabolized by CYP4F enzymes, and a terminal half-life suitable for once-daily dosing.
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| Toxicity/Toxicokinetics |
No specific toxicity data was found. S1P1 agonists in clinical use (e.g., fingolimod) have known side effects including bradycardia (first-dose effect), macular edema, elevated liver enzymes, and increased risk of infections. S1P1 agonist 1 has been reported to have little effect on heart rate and a smaller decrease in blood pressure compared to FTY720, suggesting a potentially improved cardiovascular safety profile. However, formal toxicology studies are not reported.
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| References |
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| Additional Infomation |
S1P1 agonist 1 (Icanbelimod, CAS: 1220973-37-4 for the free base; active salt CAS: 1514888-56-2) has molecular formula C22H22ClN3O5 and molecular weight 443.88. It is a potent and orally active S1P receptor agonist that induces S1P1 internalization (EC50 = 9.83 nM). It attenuates EAE disease in rat models of multiple sclerosis. For research use only.
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| CAS # |
1220973-37-4
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| Appearance |
White to off-white solid powder
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ≥ 125 mg/mL (~281.61 mM)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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