NPS-2143_hydrochloride_DataSheet_MedChemExpress

Inhibitors, Agonists, Screening Libraries Data SheetBIOLOGICAL ACTIVITY:NPS–2143(SB 262470A ) is a selective potent calcium ion–sensing receptor antagonist with IC50 of 43 and 41 nM for cytoplasmic Ca2+ concentrations and parathyroid hormone secretion, respectively.IC50 value: 43 nM(for Ca2+ receptor) [1]Target: CaSRin vitro: NPS 2143, even when tested at much higher concentrations (3 microM), did not affect the activity of a number of other G protein–coupled receptors, including those most structurally homologous to the Ca2+ receptor. NPS 2143 stimulated parathyroid hormone (PTH) secretion from bovine parathyroid cells (EC50 of 41 nM) over a range of extracellular Ca2+ concentrations and reversed the effects of the calcimimetic compound NPS R–467 on [Ca2+]i and on secretion of PTH [1]. The first reported calcilytic compound was NPS 2143, an orally active molecule which elicits rapid, 3– to 4–fold increases in circulating levels of PTH [2].in vivo: When infused intravenously in normal rats, NPS 2143 caused a rapid and large increase in plasma levels of PTH. Ca2+ receptor antagonists are termed calcilytics and NPS 2143 is the first substance (either atomic or molecular) shown to possess such activity [1].When administered together with an antiresorptive agent (estradiol), NPS 2143 causes an increase in trabecular bone volume and bone mineral density in osteopenic rats [2].PROTOCOL (Extracted from published papers and Only for reference)Cell assay [6]The effect of NPS 2143 on Ca2+ receptor–dependent regulation of PTH secretion and cyclic AMP formation was assessed using primary cultures of dissociated bovine parathyroid cells. Following overnight culture, the cells were removed from the flasks by decanting and washed with buffer containing 126 mM NaCl, 4 mM KCl, 1 mM MgSO4, 0.5 mM CaCl2, 0.7 mM K2HPO4/KH2PO4, 20mM Na–HEPES, pH 7.45, 1 mg/ml glucose, and 0.1% bovine serum albumin. Portions (0.2 ml) of this cellular suspension were added to 12– × 75–mm polystyrene tubes with or without NPS 2143 and/or varying concentrations of CaCl2. Incubations (in triplicate) at 37°C for 20 or 30 min were terminated by placing the tubes on ice. Cells were pelleted by centrifugation (500g for 10 min at 4°C) and 0.1 ml of supernatant was immediately assayed for PTH content. A portion of the cellular suspension was left on ice during the incubation period and then processed in parallel with the other tubes. The amount of PTH in the supernatant from the tubes maintained on ice was defined as "basal release" and was subtracted from all other samples. PTH levels were quantified using a rat PTH(1–34)immunoradiometric assay kit, which also detects bovine PTH. For each experiment, results were expressed as picograms of PTH released/106 cells and then normalized to PTH released in 0.5 mM Ca2+. Cell numbers were determined by counting nuclei in a hemocytometer after lysing the cells and staining the nuclei with cresyl violet.Animal administration [6]On the day of study, the rats were infused intravenously (0.1 ml/kg . min) for 120 min with NPS 2143 (0.1 μmol/kg . min) or vehicle, aProduct Name:NPS–2143 (hydrochloride)Cat. No.:HY-10171CAS No.:324523-20-8Molecular Formula:C 24H 26Cl 2N 2O 2Molecular Weight:445.38Target:CaSR Pathway:GPCR/G Protein Solubility:10 mM in DMSO20% aqueous solution of 2–hydroxypropyl–β–cyclodextrin. Blood samples (0.5 ml) were collected before and at various times after the start of the infusion for measurements of plasma levels of PTH and Ca2+. To prevent excessive blood volume loss during the course of the experiment, for each blood sample the erythrocyte pellet was resuspended in an equal volume of normal rat plasma and reinjected. Plasma levels of Ca2+ were measured immediately after collection using a model 634 ionized calcium analyzer. PTH levels were measured using the Immutopics rat PTH(1–34) immunoradiometric assay kit.References:[1]. Huang Y, Breitwieser GE. Rescue of calcium–sensing receptor mutants by allosteric modulators reveals a conformational checkpoint in receptor biogenesis. J Biol Chem. 2007 Mar 30;282(13):9517–25[2]. Marquis, Robert W.; Lago, Amparo M.; Callahan, James F.;Antagonists of the Calcium Receptor. 2. Amino Alcohol–Based Parathyroid Hormone Secretagogues. Journal of Medicinal Chemistry (2009), 52(21), 6599–6605.[3]. Yamamura A, Guo Q, Yamamura H, Zimnicka AM, Pohl NM, Smith KA, Fernandez RA, Zeifman A, Makino A, Dong H, Yuan JX.Enhanced Ca2+–sensing receptor function in idiopathic pulmonary arterial hypertension.Circ Res. 2012 Aug 3;111(4):469–81. Epub 2012 Jun 22.[4]. Nakajima S, Hira T, Hara H.Calcium–sensing receptor mediates dietary peptide–induced CCK secretion in enteroendocrine STC–1 cells.Mol Nutr Food Res. 2012 May;56(5):753–60.[5]. Davey AE, Leach K, Valant C, Conigrave AD, Sexton PM, Christopoulos A.Positive and negative allosteric modulators promote biased signaling at the calcium–sensing receptor.Endocrinology. 2012 Mar;153(3):1232–41. Epub 2011 Dec 30.[6]. Nemeth EF, et al. Calcilytic compounds: potent and selective Ca2+ receptor antagonists that stimulate secretion of parathyroid hormone. J Pharmacol Exp Ther. 2001 Oct;299(1):323–31.Caution: Product has not been fully validated for medical applications. 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