UT-B-IN-1 (UTBINH-14) is a reversible, competitive and selective urea transporter B (UT-B) inhibitor (antagonist) with IC50 of UT-B in humans and mice respectively. 10 and 25 nM. UT-B-IN-1 has low toxicity and high selectivity compared to the UT-A isoform. UT-B-IN-1 increases urine output and reduces urinary permeability in mice. UT-B-IN-1 may be utilized to study the diuretic mechanism.

Physicochemical Properties

Molecular Formula	C20H17N5O2S3
Molecular Weight	455.576279401779
Exact Mass	455.054
CAS #	892742-76-6
PubChem CID	16019476
Appearance	White to off-white solid powder
LogP	4.3
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	6
Heavy Atom Count	30
Complexity	697
Defined Atom Stereocenter Count	0
SMILES	S(C1C([H])=C([H])C(C([H])([H])C([H])([H])[H])=C([H])C=1[H])(C1=C2N=C(C3=C(C([H])=C([H])S3)N2N=N1)N([H])C([H])([H])C1=C([H])C([H])=C([H])S1)(=O)=O
InChi Key	UYFZCWXRMHSLTC-UHFFFAOYSA-N
InChi Code	InChI=1S/C20H17N5O2S3/c1-2-13-5-7-15(8-6-13)30(26,27)20-19-22-18(21-12-14-4-3-10-28-14)17-16(9-11-29-17)25(19)24-23-20/h3-11H,2,12H2,1H3,(H,21,22)
Chemical Name	10-(4-ethylphenyl)sulfonyl-N-(thiophen-2-ylmethyl)-5-thia-1,8,11,12-tetrazatricyclo[7.3.0.02,6]dodeca-2(6),3,7,9,11-pentaen-7-amine
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

Targets	IC50: 10 nM (human UT-B); 25 nM (mouse UT-B)[1]
ln Vitro	With an IC50 value of 26.7 nM, UT-B-IN-1 (1-1000 nM) inhibits the efflux of urea from red blood cells that have been primed with urea [1]. Water transport in erythrocytes lacking AQP1 is inhibited by UT-B-IN-1 [1]. For MDCK cells, UT-B-IN-1 (0–10 μM; 24 h) is not cytotoxic [1].
ln Vivo	In mice with free access to food and water, UT-B-IN-1 (300 μg; i.p., 1x) increases urine production and decreases urine permeability[1].
Cell Assay	Cell Cytotoxicity Assay[1] Cell Types: Madin-Darby canine kidney (MDCK) cell line Tested Concentrations: 0-10 μM Incubation Duration: 24 hrs (hours) Experimental Results: demonstrated no cytotoxic effect to MDCK cells.
Animal Protocol	Animal/Disease Models: Male mice (wild-type or UT-B knockout)[1] Doses: 300 μg Route of Administration: intraperitoneal (ip) injection, 300 μg, once Experimental Results: decreased urine osmolality and urea concentration in wild-type mice with V2-receptor agonist dDAVP injection. Increased urine volume and decreased urine osmolality and urea concentration in mice with free access to food and water but without dDAVP.
References	[1]. Triazolothienopyrimidine inhibitors of urea transporter UT-B reduce urine concentration. J Am Soc Nephrol. 2012 Jul;23(7):1210-20.

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	2.1950 mL	10.9750 mL	21.9500 mL
	5 mM	0.4390 mL	2.1950 mL	4.3900 mL
	10 mM	0.2195 mL	1.0975 mL	2.1950 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.