From UTP to AR-C118925, the discovery of a potent non nucleotide antagonist of the P2Y2 receptor

Kindon, Nicholas D.; Davis, A. M.; Dougall, Iain G.; Dixon, J. Michael; Johnson, Timothy A.; Walters, Iain A. S.; Thom, Steve; McKechnie, K.; Meghani, Premji; Stocks, Michael J.

doi:10.1016/j.bmcl.2017.09.043

Cited by 26 publications

(33 citation statements)

References 24 publications

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“… 7 To date, the only reported high-affinity P2Y 2 R antagonists were those developed by scientists from AstraZeneca resulting in the non-nucleotide P2Y 2 R antagonist AR-C118925 ( 1 ). 8 , 9 …”

Section: Introductionmentioning

confidence: 99%

Synthesis and Evaluation of the First Fluorescent Antagonists of the Human P2Y₂ Receptor Based on AR-C118925

et al. 2018

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The human P2Y2 receptor (hP2Y2R) is a G-protein-coupled receptor that shows promise as a therapeutic target for many important conditions, including for antimetastatic cancer and more recently for idiopathic pulmonary fibrosis. As such, there is a need for new hP2Y2R antagonists and molecular probes to study this receptor. Herein, we report the development of a new series of non-nucleotide hP2Y2R antagonists, based on the known, non-nucleotide hP2Y2R antagonist AR-C118925 (1), leading to the discovery of a series of fluorescent ligands containing different linkers and fluorophores. One of these conjugates, 98, displayed micromolar affinity for hP2Y2R (pKd = 6.32 ± 0.10, n = 17) in a bioluminescence-energy-transfer (BRET) assay. Confocal microscopy with this ligand revealed displaceable membrane labeling of astrocytoma cells expressing untagged hP2Y2R. These properties make 98 one of the first tools for studying hP2Y2R distribution and organization.

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Section: Introductionmentioning

confidence: 99%

Synthesis and Evaluation of the First Fluorescent Antagonists of the Human P2Y₂ Receptor Based on AR-C118925

et al. 2018

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“…Applying the Cheng–Prusoff equation generates a K B for AR‐C118925XX of approximately 13 nM, which is closer to the value calculated here. Shortly afterwards, a group from AstraZeneca described the original design and synthesis of AR‐C118925XX and reported pA 2 and K B values of 7.8 and 15.8 nM, respectively, at hP2Y 2 receptors expressed in Jurkat cells (Kindon et al, ). Neither the Schild plot nor the slope of the plot was included, however, so the mode of antagonism cannot be confirmed.…”

Section: Discussionmentioning

confidence: 99%

“…The biggest methodological difference between our study and those of Rafehi, Burbiel, et al (2017), Rafehi, Neumann, et al (2017), and Kindon et al (2017) is that they used multi-well plates and a microplate reader to record changes in cytoplasmic Ca 2+ levels. To generate a CRC, multiple populations of cells were stimulated once only, with a single concentration of UTP.…”

Section: Mode Of Action Of Ar-c118925xxmentioning

confidence: 99%

Characterisation of P2Y₂ receptors in human vascular endothelial cells using AR‐C118925XX, a competitive and selective P2Y₂ antagonist

Muoboghare

Drummond

Kennedy

2019

British J Pharmacology

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Background and Purpose There is a lack of potent, selective antagonists at most subtypes of P2Y receptor. The aims of this study were to characterise the pharmacological properties of the proposed P2Y2 receptor antagonist, AR‐C118925XX, and then to use it to determine the role of P2Y2 receptors in the action of the P2Y2 agonist, UTP, in human vascular endothelial cells. Experimental Approach Cell lines expressing native or recombinant P2Y receptors were superfused constantly, and agonist‐induced changes in intracellular Ca2+ levels monitored using the Ca2+‐sensitive fluorescent indicator, Cal‐520. This set‐up enabled full agonist concentration–response curves to be constructed on a single population of cells. Key Results UTP evoked a concentration‐dependent rise in intracellular Ca2+ in 1321N1‐hP2Y2 cells. AR‐C118925XX (10 nM to 1 μM) had no effect per se on intracellular Ca2+ but shifted the UTP concentration–response curve progressively rightwards, with no change in maximum. The inhibition was fully reversible on washout. AR‐C118925XX (1 μM) had no effect at native or recombinant hP2Y1, hP2Y4, rP2Y6, or hP2Y11 receptors. Finally, in EAhy926 immortalised human vascular endothelial cells, AR‐C118925XX (30 nM) shifted the UTP concentration–response curve rightwards, with no decrease in maximum. Conclusions and Implications AR‐C118925XX is a potent, selective and reversible, competitive P2Y2 receptor antagonist, which inhibited responses mediated by endogenous P2Y2 receptors in human vascular endothelial cells. As the only P2Y2‐selective antagonist currently available, it will greatly enhance our ability to identify the functions of native P2Y2 receptors and their contribution to disease and dysfunction.

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“…AR‐C118925 (5‐[[5‐(2,8‐dimethyl‐5 H ‐dibenzo[a,d]cyclohepten‐5‐yl)‐3,4‐dihydro‐2‐oxo‐4‐thioxo‐1(2 H )‐pyrimidinyl]methyl]‐ N ‐2 H ‐tetrazol‐5‐yl‐2‐furancarboxamide, 16 ) is a potent and selective antagonist for P2Y 2 receptors (Kindon et al, 2017; Rafehi, Burbiel, Attah, Abdelrahman, & Müller, 2017; Rafehi, Neumann, et al, 2017).…”

Section: Selective Ligand Tools To Study P2y Receptorsmentioning

confidence: 99%

Update of P2Y receptor pharmacology: IUPHAR Review 27

Jacobson

Delicado

Gachet

et al. 2020

British J Pharmacology

180

184

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Eight G protein-coupled P2Y receptor subtypes respond to extracellular adenine and uracil mononucleotides and dinucleotides. P2Y receptors belong to the δ group of rhodopsin-like GPCRs and contain two structurally distinct subfamilies: P2Y 1 , P2Y 2 , P2Y 4 , P2Y 6 , and P2Y 11 (principally G q protein-coupled P2Y 1 -like) and P2Y 12-14 (principally G i protein-coupled P2Y 12 -like) receptors. Brain P2Y receptors occur in neurons, glial cells, and vasculature. Endothelial P2Y 1 , P2Y 2 , P2Y 4 , and P2Y 6 receptors induce vasodilation, while smooth muscle P2Y 2 , P2Y 4 , and P2Y 6 receptor activation leads to vasoconstriction. Pancreatic P2Y 1 and P2Y 6 receptors stimulate while P2Y 13 receptors inhibits insulin secretion. Antagonists of P2Y 12 receptors, and potentially P2Y 1 receptors, are anti-thrombotic agents, and a P2Y 2 /P2Y 4 receptor agonist treats dry eye syndrome in Asia. P2Y receptor agonists are generally pro-inflammatory, and antagonists may eventually treat inflammatory conditions. This article reviews recent developments in P2Y receptor pharmacology (using synthetic agonists and antagonists), structure and biophysical properties (using X-ray crystallography, mutagenesis and modelling), physiological and pathophysiological roles, and present and potentially future therapeutic targeting.Abbreviations: BMD, bone mineral density; DUSP, dual specificity protein phosphatase; ECL, extracellular loop; EPAC, exchange protein activated by cAMP; KO, knockout; MSD, musculoskeletal disorder; SNP, single nucleotide polymorphism; SS, Sjögren's syndrome; TM, transmembrane helix.

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From UTP to AR-C118925, the discovery of a potent non nucleotide antagonist of the P2Y2 receptor

Cited by 26 publications

References 24 publications

Synthesis and Evaluation of the First Fluorescent Antagonists of the Human P2Y₂ Receptor Based on AR-C118925

Synthesis and Evaluation of the First Fluorescent Antagonists of the Human P2Y₂ Receptor Based on AR-C118925

Characterisation of P2Y₂ receptors in human vascular endothelial cells using AR‐C118925XX, a competitive and selective P2Y₂ antagonist

Update of P2Y receptor pharmacology: IUPHAR Review 27

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From UTP to AR-C118925, the discovery of a potent non nucleotide antagonist of the P2Y2 receptor

Cited by 26 publications

References 24 publications

Synthesis and Evaluation of the First Fluorescent Antagonists of the Human P2Y2 Receptor Based on AR-C118925

Synthesis and Evaluation of the First Fluorescent Antagonists of the Human P2Y2 Receptor Based on AR-C118925

Characterisation of P2Y2 receptors in human vascular endothelial cells using AR‐C118925XX, a competitive and selective P2Y2 antagonist

Update of P2Y receptor pharmacology: IUPHAR Review 27

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Synthesis and Evaluation of the First Fluorescent Antagonists of the Human P2Y₂ Receptor Based on AR-C118925

Synthesis and Evaluation of the First Fluorescent Antagonists of the Human P2Y₂ Receptor Based on AR-C118925

Characterisation of P2Y₂ receptors in human vascular endothelial cells using AR‐C118925XX, a competitive and selective P2Y₂ antagonist