135PAM1 increases the intracellular Ca²⁺ response to amidated, but not free acid RXFP3 agonists in cells coexpressing RXFP3 and G_qI5.

<p>Intracellular Ca²⁺ responses by HEK-293 cells coexpressing RXFP3 and G_qI5 were measured in response to escalating concentrations of 135PAM1 using probe (EC₂₀) concentrations of Relaxin-3_NH2 (A), Relaxin-3_OH (B), R3/I5_NH2 (C), or R3/I5_OH (D).</p

Structure of 135PAM1 (3-[3,5-Bis(trifluoromethyl)phenyl]-1-(3,4-dichlorobenzyl)-1-[2-(5-methoxy-1H-indol-3-yl)ethyl]urea).

<p>Structure of 135PAM1 (3-[3,5-Bis(trifluoromethyl)phenyl]-1-(3,4-dichlorobenzyl)-1-[2-(5-methoxy-1H-indol-3-yl)ethyl]urea).</p

Shifting of amidated RXFP3 agonist concentration response curves by 135PAM1 in cells lacking chimeric G proteins.

<p>SK-N-MC cells coexpressing RXFP3 and a reporter construct linking CRE activity to β-galactosidase were incubated with fixed concentrations of 135PAM1 (0, 0.2, 2 and 20 µM) and increasing concentrations of Relaxin-3_NH2 (A) or R3I5_NH2 (B).</p

135PAM1 shifts the concentration response curves of Relaxin-3_NH2 and R3/I5_NH2.

<p>HEK-293 cells coexpressing RXFP3 and G_qI5 were incubated with fixed concentrations of 135PAM1 (0, 0.2, 2 and 20 µM) 10 min before the addition of increasing concentrations of Relaxin-3_NH2 (A), R3I5_NH2 (B), Relaxin-3_OH (C) or R3I5_OH (D).</p

135PAM1 lacks affinity at the orthosteric binding site of RXFP3 receptor.

<p>135PAM1 did not displace [125I] R3/I5_NH2 at concentrations of up to 20 µM, but instead increased total binding. R3/I5_NH2 displaced the tracer with a pIC₅₀ of 8.76 (8.91 to 8.61).</p

Identification and SAR of Glycine Benzamides as Potent Agonists for the GPR139 Receptor

A focused high throughput screening for GPR139 was completed for a select 100K compounds, and new agonist leads were identified. Subsequent analysis and structure–activity relationship studies identified (<i>S</i>)-3-chloro-<i>N</i>-(2-oxo-2-((1-phenylethyl)­amino)­ethyl)­benzamide <b>7c</b> as a potent and selective agonist of hGPR139 with an EC₅₀ = 16 nM. The compound was found to cross the blood–brain barrier and have good drug-like properties amenable for oral dosing in rat

Novel Octahydropyrrolo[3,4‑<i>c</i>]pyrroles Are Selective Orexin‑2 Antagonists: SAR Leading to a Clinical Candidate

The preclinical characterization of novel octahydro­pyrrolo­[3,4-<i>c</i>]­pyrroles that are potent and selective orexin-2 antagonists is described. Optimization of physicochemical and DMPK properties led to the discovery of compounds with tissue distribution and duration of action suitable for evaluation in the treatment of primary insomnia. These selective orexin-2 antagonists are proven to promote sleep in rats, and this work ultimately led to the identification of a compound that progressed into human clinical trials for the treatment of primary insomnia. The synthesis, SAR, and optimization of the pharmacokinetic properties of this series of compounds as well as the identification of the clinical candidate, JNJ-42847922 (<b>34</b>), are described herein