An Agonist Radioligand for the Proinflammatory Lipid-Activated G Protein-Coupled Receptor GPR84 Providing Structural Insights.
Meryem KöseThanigaimalai PillaiyarVigneshwaran NamasivayamElisabetta De FilippoKatharina SylvesterTrond UlvenIvar von KügelgenChrista Elisabeth MüllerPublished in: Journal of medicinal chemistry (2019)
The orphan G protein-coupled receptor (GPCR) GPR84 is expressed on immune cells mediating proinflammatory and immunostimulatory effects. In this study, we prepared the fully efficacious, nonbiased GPR84 agonist 6-hexylamino-2,4(1H,3H)-pyrimidinedione (6) in tritium-labeled form ([3H]PSB-1584) by hydrogenation of a hexenyl-substituted precursor with tritium gas. The radioligand was characterized by kinetic, saturation, and competition assays using membranes of Chinese hamster ovary cells recombinantly expressing the human GPR84. [3H]6 reversibly labeled the receptor with high affinity (KD 2.08 nM). Structurally diverse orthosteric and allosteric ligands, including newly designed and synthesized compounds, were studied in competition binding assays. A homology model of GPR84 was generated to perform docking studies rationalizing the experimental data. The radioligand was additionally used for labeling GPR84 in native cells and tissues. [3H]6 constitutes the first GPR84 agonist radioligand representing a powerful tool for this poorly investigated GPCR, which has potential as a future drug target.
Keyphrases
- fatty acid
- induced apoptosis
- cell cycle arrest
- endothelial cells
- gene expression
- small molecule
- pet imaging
- cell death
- molecular dynamics
- oxidative stress
- machine learning
- climate change
- protein protein
- electronic health record
- molecular dynamics simulations
- transcription factor
- artificial intelligence
- ionic liquid
- mass spectrometry
- single molecule
- room temperature
- solid state