TC-SP 14 (compound 14) is an orally bioactive S1P1 agonist (EC50 = 0.042 μM) with very low activity at S1P3 (EC50 = 3.47 μM). TC-SP 14 significantly reduces blood lymphocyte counts and attenuates delayed-type hypersensitivity (DTH) to antigen challenge.

Physicochemical Properties

Molecular Formula	C25H20N2O2F2S
Molecular Weight	450.5003
Exact Mass	450.121
CAS #	1257093-40-5
PubChem CID	51346934
Appearance	Typically exists as solid at room temperature
LogP	5.286
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	6
Heavy Atom Count	32
Complexity	664
Defined Atom Stereocenter Count	0
SMILES	C1C(CN1CC2=CC(=C(C=C2)C3=NC4=C(S3)C=CC(=C4)CC5=CC=CC=C5F)F)C(=O)O
InChi Key	GVXGVDIXINMAAL-UHFFFAOYSA-N
InChi Code	InChI=1S/C25H20F2N2O2S/c26-20-4-2-1-3-17(20)9-15-6-8-23-22(11-15)28-24(32-23)19-7-5-16(10-21(19)27)12-29-13-18(14-29)25(30)31/h1-8,10-11,18H,9,12-14H2,(H,30,31)
Chemical Name	1-[[3-fluoro-4-[5-[(2-fluorophenyl)methyl]-1,3-benzothiazol-2-yl]phenyl]methyl]azetidine-3-carboxylic acid
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	TC-SP 14 (Compound 14) is non-mutagenic, does not appreciably block hERG channels, and neither stimulates nor inhibits human cytochrome P450 enzymes [1].
ln Vivo	Twenty-four hours after administration, TC-SP 14 (Compound 14) (0–3 mg/kg, PO, once) causes a dose-dependent drop in circulating lymphocytes [1]. Ovalbumin (OVA)-induced ear swelling is significantly reduced by TC-SP 14 (0-3 mg/kg), taken orally once daily for 10 days [1]. TC-SP 14 has acceptable properties (2–15 mg/kg IV or PO once) [1]. Pharmacokinetic characteristics of TC-SP 14 in male cynomolgus monkeys and female Sprague-Dawley rats [1]. NHP CL (L/h/kg) = 0.33 0.50 Vss (L/kg) 3.3 1.6 T1/2 (h) 7.5 35.2 MRT (h) 10.3% F 68 23 Species Rat
Animal Protocol	Animal/Disease Models: Lewis rat (female, n = 5)/group) [1] Doses: 0.3, 1.0 and 3.0 mg/kg Route of Administration: po (po (oral gavage)) once Experimental Results: There was a dose-dependent decrease in circulating blood lymphocytes 24 hrs (hrs (hours)) after administration, resulting in a dose-dependent reduction of circulating blood lymphocytes at 3.0 mg/kg Maximal lymphopenia (percent reduction of 74 lymphocytes relative to vehicle). Animal/Disease Models: OVA-immunized Lewis rats (female, n = 8/group) [1] Doses: 0.1, 0.3, 1.0, and 3.0 mg/kg Route of Administration: Orally, one time/day for 10 days Experimental Results: OVA-induced Ear swelling is Dramatically diminished at doses of 0.3 mg/kg or higher. Animal/Disease Models: Female SD (SD (Sprague-Dawley)) rats, male cynomolgus monkeys (NHP (non-human primates)) (n=3/group) [1] Doses: 2 (IV, rat), 4 (IV, NHP), 10 (PO, NHP), 15 mg/kg (PO, rat) Route of Administration: IV, PO, once (pharmacokinetic/PK/PK analysis) Experimental Results: Has acceptable properties, exhibits low clearance rate, moderate steady-state volume of distribution, moderate to long mean residence t
References	[1]. Discovery of a Potent, S1P3-Sparing Benzothiazole Agonist of Sphingosine-1-Phosphate Receptor 1 (S1P1). ACS Med Chem Lett. 2010 Nov 9;2(2):102-6.

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	2.2198 mL	11.0988 mL	22.1976 mL
	5 mM	0.4440 mL	2.2198 mL	4.4395 mL
	10 mM	0.2220 mL	1.1099 mL	2.2198 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.