化合物信息

Description

NPS 2143 is a novel potent and selective antagonist of Ca(2+) receptor with IC50 of 43 nM in HEK 293 cells.

Targets

IC₅₀

In Vitro

NPS 2143 blocks increases in cytoplasmic Ca²⁺ concentrations with IC50 of 43 nM elicited by activating the Ca²⁺ receptor in HEK 293 cells expressing the human Ca²⁺ receptor. ^[1] NPS 2143 stimulates parathyroid hormone (PTH) secretion from bovine parathyroid cells with EC50 of 41 nM. Moreover, NPS 214 also blocks the inhibitory effects of calcimimetic NPS R-467 on PTH secretion from bovine parathyroid cells and the inhibitory effects of extracellular Ca2+ on isoproterenol-stimulated increases in cyclic AMP formation. ^[1] In HEK-293 cells transiently expressing hCaSRs, NPS 2143 significantly suppresses the kokumi taste by effectively inhibiting the activity of both GSH (data not shown) and γ-Glu-Val-Gly. ^[3] A recent study shows that NPS 2143 treatment suppresses low molecular weight fractions of azuki hydrolysate-induced cholecystokinin (CCK) secretion in CaSR-transfected HEK 293 cells. ^[4]

In Vivo

In rats, NPS 2143 results in a rapid 4- to 5-fold increase in plasma PTH levels and also a transient increase in plasma Ca²⁺ levels. ^[1] In normotensive rats, NPS 2143 administration (1 mg/kg) by i.v. markedly increases mean arterial blood pressure (MAP) in the presence of parathyroid glands. ^[2]

Clinical Trials

Features

Assays for Assessing Potency and Selectivity of Compounds on Ca2+ Receptor

This clonal cell line, referred to as HEK 293 4.0-7 cells, are used in a high-throughput screening format to detect agonists and allosteric activators of the Ca²⁺ receptor. Changes in the concentration of cytoplasmic Ca²⁺([Ca2+]i) provide a quantitative and functional assessment of Ca²⁺receptor activity in these cells and the results using this assay parallel those obtained using a homologous expression system of bovine parathyroid cells. On-line continuous measurements of fluorescence in fluo-3- or fura-2-loaded HEK 293 4.0-7 cells are obtained using a custom-built spectrofluorimeter or a fluorescence imaging plate reader instrument. NPS 2143 is incubated with cells for 1 minute before increasing the concentration of extracellular Ca²⁺ from 1.0 mM to 1.75 mM. NPS 2143 is tested individually at a concentration of 100 μg/ml (20 μM–80 μM) and those causing more than a 40% inhibition of the control response are considered to be biologically active. To determine the potencies (IC50) of NPS 2143 with biological activity, concentration-response curves are obtained and then, as an initial assessment of selectivity, the effects of NPS 2143 on [Ca²⁺]i evoked by other G protein-coupled receptors are examined at a concentration several times their IC50. Wild-type HEK 293 cells (and HEK 293 4.0-7 cells) express receptors for thrombin, bradykinin, and ATP, which couple to the mobilization of intracellular Ca²⁺. These responses can be studied to quickly assess any nonselective action of compounds on G protein-coupled receptors. Additional assays for selectivity include HEK 293 cells engineered to express receptors most homologous in sequence and topology to the Ca2+ receptor. These include native or chimeric receptors for various metabotropic glutamate and γ-aminobutyric acid type B receptors (GABABRs). Chimeric receptors are created using partial sequences of metabotropic glutamate receptors and Ca²⁺ receptors, engineered to couple to activation of phospholipase C and release of intracellular Ca^{2+ in HEK 293 cells. NPS 2143 lacking pan-activity are then subjected to structural modifications and their potencies and selectivities monitored using these HEK 293 4.0-7 cell assays in an iterative process.}

Animal Models

Chronic indwelling catheters are implanted in the inferior vena cava and in the abdominal aorta of male Sprague-Dawley rats.

Formulation

NPS 2143 is dissolved in 20% aqueous solution of 2-hydroxypropyl-β-cyclodextrin.

Doses

≤0.1 μmol/kg · min

Administration

Administered via i.v.