Cell lines

HEK293-T cells expressing zebrafish GPR139

Preparation Method

The cells were serum starved in the media with 0.5% FBS for 18-20h, and then treated with the vehicle (control) or various concentrations of JNJ-63533054 in the media for 6h.

Reaction Conditions

4-128nM in 0.5% DMSO, 6h

Applications

JNJ-63533054 induced luciferase activity against zebrafish GPR139 in a concentration-dependent manner. For zebrafish GPR139, the maximal induction of 3.3-fold of vehicle control was achieved at a concentration of 128nM. From the dose-response curve, the half effective maximal concentration (EC₅₀) values of JNJ-63533054 to zebrafish GPR139 was 3.91nM.

Animal models

Male Sprague-Dawley rats, 350-450g

Preparation Method

Separate groups of rats were orally dosed at 2 hours into the light phase with the GPR139 agonist JNJ-63533054 (3, 10, and 30mg/kg, n=8), its less active enantiomer JNJ-63770044 (10mg/kg, n=7), and L-Trp (200mg/kg, n=7) or vehicle (0.5% hydroxypropyl methylcellulose in suspension) administered in the same animals receiving each dose.

Dosage form

3, 10, and 30mg/kg, orally dosed

Applications

The results are presented for the first hour after dosing based on the short-lasting effects observed. Compared with vehicle, JNJ-63533054 induced a dose-dependent reduction in locomotor activity in the first hour after the treatment that reached significance from the dose of 10mg/kg onward. In contrast, at the same dose of 10mg/kg, its less active enantiomer did not modify locomotor activity. For comparison, L-Trp was tested in the same experimental conditions and spontaneous locomotor activity was not affected at 200mg/kg.

JNJ-63533054 is a potent and selective agonist of hGPR139 with an EC₅₀= 16nM^[1,2]. JNJ-63533054 is an excellent candidate to explore the unknown in vivo function of central GPR139^[3]

JNJ-63533054 specifically activated human GPR139 in the calcium mobilization (EC₅₀=16±6nM) and GTPγS binding (EC₅₀=17±4nM) assays^[1]. JNJ-63533054 effectively binds and acts as an agonist to zebrafish GPR139 with EC₅₀of 3.91nM^[4]

JNJ-63533054 activated the rat and mouse GPR139 receptor with similar potency (rat EC₅₀=63±24nM, mouse EC₅₀=28±7nM)^[1]. JNJ-63533054(oral dose of 10mg/kg) crossed the blood-brain barrier in both male mice and male rats, and the brain to plasma ratio was close to 1 in mouse and slightly higher in rat. In the marble burying test, JNJ-63533054(10mg/kg p.o.) produced a small anxiolytic-like effect, with no interaction with fluoxetine, and no effect in elevated plus maze (EPM)^[3]. Administration of a high dose (1μg/g BW) of JNJ-6353305 had no effect on locomotor activity, and fear response, but fear-conditioned place avoidance was diminished; zebrafish treated with a lower dose(0.1μg/g BW) of GPR139 agonist exhibited avoidance to the contextual compartments^[4]. JNJ-63533054(oral dose of 3-30mg/kg) dose-dependently reduced non-rapid eye movement (NREM) latency and increased NREM sleep duration without altering rapid eye movement (REM) sleep when acutely administered at the beginning of the light phase. This effect progressively dissipated upon 7-day repeated dosing^[5]. Systemic administration of JNJ-63533054 (30mg/kg, p.o.) reversed compulsive-like alcohol drinking and decreases withdrawal-induced hyperalgesia in alcohol-dependent rats that exhibit symptoms of alcohol dependence^[6]

References:
[1]. Liu C, Bonaventure P, et al. 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;88(5):911-925.
[2]. Dvorak CA, Coate H, et al. Identification and SAR of Glycine Benzamides as Potent Agonists for the GPR139 Receptor. ACS Med Chem Lett. 2015;6(9):1015-1018. Published 2015 Jul 20.
[3]. Shoblock JR, Welty N, et al. In vivo Characterization of a Selective, Orally Available, and Brain Penetrant Small Molecule GPR139 Agonist. Front Pharmacol. 2019;10:273. Published 2019 Mar 21.
[4]. Roy N, Ogawa S, et al. Habenula GPR139 is associated with fear learning in the zebrafish. Sci Rep. 2021;11(1):5549. Published 2021 Mar 10.
[5]. Wang L, Dugovic C, et al. Putative role of GPR139 on sleep modulation using pharmacological and genetic rodent models. Eur J Pharmacol. 2020;882:173256.
[6]. Kononoff J, Kallupi M, et al. Systemic and Intra-Habenular Activation of the Orphan G Protein-Coupled Receptor GPR139 Decreases Compulsive-Like Alcohol Drinking and Hyperalgesia in Alcohol-Dependent Rats. eNeuro. 2018;5(3):ENEURO.0153-18.2018. Published 2018 Jul 2.