S1P1 agonist 5 is a selective and orally bioactive S1P1 agonist. S1P1 agonist 5 inhibits the egress of lymphocytes from lymphoid tissue into the peripheral blood. S1P1 agonist 5 may be used for studying multiple sclerosis (MS).

Physicochemical Properties

Molecular Formula	C23H24CLN2NAO4
Molecular Weight	450.89
CAS #	2760666-20-2
Appearance	Typically exists as solid at room temperature
HS Tariff Code	2934.99.9001
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)

Biological Activity

ln Vitro	Compound 21l, or S1P1 agonist 5, exhibits remarkable in vitro efficacies, demonstrating EC50s for β-arrestin recruitment and internalization of 7.03 nM and 11.8 nM, respectively[1].
ln Vivo	Oral bioavailability of S1P1 agonist 5 is demonstrated in rats (F=54.2%) and dogs (F=31.8%)[1]. Within a day, lymphopenia can be recovered from when S1P1 agonist 5 (10 mg/kg; po) prevents lymphocyte egress from lymphoid tissue to the peripheral blood[1]. In EAE mice, S1P1 agonist 5 (3, 10 mg/kg, po, once daily for 20 days) reduces the severity of the disease and its overall course, exhibiting positive drug-like characteristics[1]. S1P1 pharmacokinetic parameters in male beagle dogs and rats[1]. Rat dog iv T1/2 (h) 1.4±0.3 5.70±1.2 AUC0-∞ (ng*h/mL) 931.3±95.7 14,830.8±5475.4 CL (mL/min/kg) 17.6±2.0 149.9±62.5 Vss (L/kg) 1.7±0.2 828.7±134.2 po Cmax (ng/mL) 1661.1±916.6 3979.4±483.5 Tmax(h) 0.9±0.8 1.3±0.5 T1 /2 (h) 1.4±0.2 4.9±0.6 AUC0-∞ (ng*h/mL) 5044.9±1061 23,109.9±7752.2 F (%) 54.2 31.8 Rats, 1 mg/kg for iv; 10 mg/kg for po. 2 mg/kg IV; 10 mg/kg for po in dogs[1]
Animal Protocol	Animal/Disease Models: rats, male beagle dogs[1] Doses: Route of Administration: 1 mg/kg for iv and 10 mg/kg for po (rats); 2 mg /kg for iv and 10 mg/kg for po(dogs) Experimental Results: demonstrated good oral bioavaliability in rats (F=54.2%) and dogs (F=31.8%). Animal/Disease Models: male wistar rats (5 week, 160-180 g)[1] Doses: 10 mg/kg Route of Administration: po Experimental Results: Inhibited the lymphocyte egress from the lymphoid tissue to the peripheral blood and that lymphopenia can be recovered within 24 hrs (hours). Animal/Disease Models: female C57BL/6 mice (10 weeks, 19−22 g) (experimental autoimmune encephalitis (EAE) mouse model)[1] Doses: 3, 10 mg/kg (dissolved in 2.5% DMSO and 5% Kolliphor HS 15 (Sigma-Aldrich) in distilled water) Route of Administration: po , one time/day, 20 days Experimental Results: Ameliorated the disease progression and overall severity in EAE mice, showing favorable drug-like properties.
References	[1]. Discovery of Novel Sphingosine-1-Phosphate-1 Receptor Agonists for the Treatment of Multiple Sclerosis. J Med Chem. 2022; 65(4):3539-3562.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.2178 mL	11.0892 mL	22.1784 mL
	5 mM	0.4436 mL	2.2178 mL	4.4357 mL
	10 mM	0.2218 mL	1.1089 mL	2.2178 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.