JNJ-63533054, a glycine benzamide analog, is a potent, selective and brain penetrant agonist of hGPR139 (human G-protein-coupled receptors) with an EC50 of 16 nM. It was found during a targeted high throughput screening for GPR139 that was carried out for a chosen 100K compounds. Following further examination and research on the structure-activity relationship, (S)-3-chloro-N-(2-oxo-2-((1-phenylethyl)amino)ethyl)benzamide, also known as JNJ-63533054, was found to be a strong and specific agonist of hGPR139, with an EC50 of 16 nM. JNJ-63533054 was found to cross the blood-brain barrier and have good drug-like properties amenable for oral dosing in rat.

Physicochemical Properties

Molecular Formula	C17H17CLN2O2
Molecular Weight	316.79
Exact Mass	316.097
Elemental Analysis	C, 64.46; H, 5.41; Cl, 11.19; N, 8.84; O, 10.10
CAS #	1802326-66-4
Related CAS #	1802326-66-4
PubChem CID	2548547
Appearance	White to off-white solid powder
Density	1.2±0.1 g/cm3
Boiling Point	559.7±45.0 °C at 760 mmHg
Flash Point	292.3±28.7 °C
Vapour Pressure	0.0±1.5 mmHg at 25°C
Index of Refraction	1.586
LogP	3.13
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	2
Rotatable Bond Count	5
Heavy Atom Count	22
Complexity	383
Defined Atom Stereocenter Count	1
SMILES	ClC1=C([H])C([H])=C([H])C(=C1[H])C(N([H])C([H])([H])C(N([H])[C@@]([H])(C([H])([H])[H])C1C([H])=C([H])C([H])=C([H])C=1[H])=O)=O
InChi Key	MWDVCHRYCKXEBY-LBPRGKRZSA-N
InChi Code	InChI=1S/C17H17ClN2O2/c1-12(13-6-3-2-4-7-13)20-16(21)11-19-17(22)14-8-5-9-15(18)10-14/h2-10,12H,11H2,1H3,(H,19,22)(H,20,21)/t12-/m0/s1
Chemical Name	3-chloro-N-[2-oxo-2-[[(1S)-1-phenylethyl]amino]ethyl]benzamide
Synonyms	JNJ63533054; JNJ 63533054; JNJ-63533054
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	human GPR139 ( Ki = 0.024 μM ); mouse GPR139 ( Ki = 0.054 μM ); rat GPR13 ( Ki = 0.075 μM )
ln Vitro	JNJ-63533054 specially induces human GPR139 in the calcium mobilization (EC50 = 16 ± 6 nM) and GTPγS binding (EC50 = 17 ± 4 nM) assays. JNJ-63533054 exhibits comparable potency in activating the GPR139 receptors in rats and mice (rat EC50 = 63 ± 24 nM, mouse EC50 = 28 ± 7 nM)[1].
ln Vivo	According to the results of an external selectivity panel comprising 50 known GPCRs, ion channels, and transporters, the small-molecule JNJ-63533054 exhibits no cross-reactivity and has appropriate pharmacokinetic characteristics for in vivo investigations. JNJ-63533054, when administered orally to rats, crosses the blood-brain barrier and reaches exposure in the micromolar range. JNJ-63533054 administration reduces rats' spontaneous locomotor activity[1].
Animal Protocol	Male Sprague-Dawley rats (350-450 g)[1] 3 mg/kg, 10 mg/kg, and 30 mg/kg Oral administration; once
References	[1]. GPR139, an Orphan Receptor Highly Enriched in the Habenula and Septum, Is Activated by the Essential Amino Acids L-Tryptophan and L-Phenylalanine. Mol Pharmacol. 2015 Nov;88(5):911-25. [2]. Identification and SAR of Glycine Benzamides as Potent Agonists for the GPR139 Receptor. ACS Med Chem Lett. 2015 Jul 20;6(9):1015-8.

Solubility Data

Solubility (In Vitro)

DMSO: ~63 mg/mL (~198.9 mM) Water: N/A Ethanol: N/A

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.89 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 (7.89 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.1567 mL	15.7833 mL	31.5667 mL
	5 mM	0.6313 mL	3.1567 mL	6.3133 mL
	10 mM	0.3157 mL	1.5783 mL	3.1567 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.