3 research outputs found

    Synthesis and Evaluation of the First Fluorescent Antagonists of the Human P2Y<sub>2</sub> Receptor Based on AR-C118925

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    The human P2Y<sub>2</sub> receptor (<i>h</i>P2Y<sub>2</sub>R) 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 <i>h</i>P2Y<sub>2</sub>R antagonists and molecular probes to study this receptor. Herein, we report the development of a new series of non-nucleotide <i>h</i>P2Y<sub>2</sub>R antagonists, based on the known, non-nucleotide <i>h</i>P2Y<sub>2</sub>R antagonist AR-C118925 (<b>1</b>), leading to the discovery of a series of fluorescent ligands containing different linkers and fluorophores. One of these conjugates, <b>98</b>, displayed micromolar affinity for <i>h</i>P2Y<sub>2</sub>R (p<i>K</i><sub>d</sub> = 6.32 ± 0.10, <i>n</i> = 17) in a bioluminescence-energy-transfer (BRET) assay. Confocal microscopy with this ligand revealed displaceable membrane labeling of astrocytoma cells expressing untagged <i>h</i>P2Y<sub>2</sub>R. These properties make <b>98</b> one of the first tools for studying <i>h</i>P2Y<sub>2</sub>R distribution and organization

    A Thieno[2,3‑<i>d</i>]pyrimidine Scaffold Is a Novel Negative Allosteric Modulator of the Dopamine D<sub>2</sub> Receptor

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    Recently, a novel negative allosteric modulator (NAM) of the D<sub>2</sub>-like dopamine receptors <b>1</b> was identified through virtual ligand screening. This ligand comprises a thieno­[2,3-<i>d</i>]­pyrimidine scaffold that does not feature in known dopaminergic ligands. Herein, we provide pharmacological validation of an allosteric mode of action for <b>1</b>, revealing that it is a NAM of dopamine efficacy and identify the structural determinants of this allostery. We find that key structural moieties are important for functional affinity and negative cooperativity, while functionalization of the thienopyrimidine at the 5- and 6-positions results in analogues with divergent cooperativity profiles. Successive compound iterations have yielded analogues exhibiting a 10-fold improvement in functional affinity, as well as enhanced negative cooperativity with dopamine affinity and efficacy. Furthermore, our study reveals a fragment-like core that maintains low μM affinity and robust negative cooperativity with markedly improved ligand efficiency

    A Thieno[2,3‑<i>d</i>]pyrimidine Scaffold Is a Novel Negative Allosteric Modulator of the Dopamine D<sub>2</sub> Receptor

    No full text
    Recently, a novel negative allosteric modulator (NAM) of the D<sub>2</sub>-like dopamine receptors <b>1</b> was identified through virtual ligand screening. This ligand comprises a thieno­[2,3-<i>d</i>]­pyrimidine scaffold that does not feature in known dopaminergic ligands. Herein, we provide pharmacological validation of an allosteric mode of action for <b>1</b>, revealing that it is a NAM of dopamine efficacy and identify the structural determinants of this allostery. We find that key structural moieties are important for functional affinity and negative cooperativity, while functionalization of the thienopyrimidine at the 5- and 6-positions results in analogues with divergent cooperativity profiles. Successive compound iterations have yielded analogues exhibiting a 10-fold improvement in functional affinity, as well as enhanced negative cooperativity with dopamine affinity and efficacy. Furthermore, our study reveals a fragment-like core that maintains low μM affinity and robust negative cooperativity with markedly improved ligand efficiency
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