GPR81 agonist 1 is a potent and selective GPR81 agonist with EC50s of 58 nM and 50 nM for human and mouse GPR81, respectively. GPR81 agonist 1 inhibits lipolysis in differentiated 3T3-L1 adipocytes. GPR81 agonist 1 inhibits lipolysis in mice without causing skin flushing. GPR81 agonist 1 is more selective for GPR81 than GPR109a.

Physicochemical Properties

Molecular Formula	C22H30N4O2S2
Molecular Weight	446.629202365875
Exact Mass	446.181
CAS #	1620992-67-7
PubChem CID	86279608
Appearance	White to off-white solid powder
LogP	3.4
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	5
Heavy Atom Count	30
Complexity	606
Defined Atom Stereocenter Count	0
SMILES	S1C(=C(C2=CC=CS2)N=C1NC(C1CCC(C)CC1)=O)CC(N1CCN(C)CC1)=O
InChi Key	ZLQAEANISSHOLB-UHFFFAOYSA-N
InChi Code	InChI=1S/C22H30N4O2S2/c1-15-5-7-16(8-6-15)21(28)24-22-23-20(17-4-3-13-29-17)18(30-22)14-19(27)26-11-9-25(2)10-12-26/h3-4,13,15-16H,5-12,14H2,1-2H3,(H,23,24,28)
Chemical Name	4-methyl-N-[5-[2-(4-methylpiperazin-1-yl)-2-oxoethyl]-4-thiophen-2-yl-1,3-thiazol-2-yl]cyclohexane-1-carboxamide
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	GPR81 Agonist 1 (Compound 2) in cell buffer (1–1000 nM) in 3T3-L1 adipocytes downstream of adipocytes [1].
ln Vivo	GPR81 agonist 1 (100 mg/kg; i.p.) inhibits cellular adipose subparts without skin flushing [1]. GPR81 Agonist 1 (10 mg/kg; i.p.) demonstrates high absorption (71%)
Animal Protocol	Animal/Disease Models: Male C57/BL6 black mouse[1] Doses: 10 mg/kg (pharmacokinetic/PK/PK analysis) Route of Administration: Ip (pharmacokinetic/PK/PK analysis) Experimental Results: demonstrated good bioavailability (71%) and Cmax (6.3 μM).
References	[1]. Identification of a novel GPR81-selective agonist that suppresses lipolysis in mice without cutaneous flushing. Eur J Pharmacol. 2014;727:1-7.

Solubility Data

Solubility (In Vitro)

DMSO : ~50 mg/mL (~111.95 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.60 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 25.0 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.60 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.2390 mL	11.1949 mL	22.3899 mL
	5 mM	0.4478 mL	2.2390 mL	4.4780 mL
	10 mM	0.2239 mL	1.1195 mL	2.2390 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.