Pharmacological characterisation of novel adenosine A
| Autor: | Kerry, Barkan, Panagiotis, Lagarias, Margarita, Stampelou, Dimitrios, Stamatis, Sam, Hoare, Dewi, Safitri, Karl-Norbert, Klotz, Eleni, Vrontaki, Antonios, Kolocouris, Graham, Ladds |
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| Rok vydání: | 2020 |
| Předmět: |
Binding Sites
Receptor Adenosine A3 Drug Evaluation Preclinical Receptors Purinergic P1 Adenosine A3 Receptor Antagonists CHO Cells Molecular Dynamics Simulation Binding Competitive Recombinant Proteins Article Rats Kinetics Radioligand Assay Structure-Activity Relationship Cricetulus Species Specificity Receptor pharmacology Cyclic AMP Mutagenesis Site-Directed Animals Humans Extracellular signalling molecules Molecular modelling |
| Zdroj: | Scientific Reports |
| ISSN: | 2045-2322 |
| Popis: | The adenosine A3 receptor (A3R) belongs to a family of four adenosine receptor (AR) subtypes which all play distinct roles throughout the body. A3R antagonists have been described as potential treatments for numerous diseases including asthma. Given the similarity between (adenosine receptors) orthosteric binding sites, obtaining highly selective antagonists is a challenging but critical task. Here we screen 39 potential A3R, antagonists using agonist-induced inhibition of cAMP. Positive hits were assessed for AR subtype selectivity through cAMP accumulation assays. The antagonist affinity was determined using Schild analysis (pA2 values) and fluorescent ligand binding. Structure–activity relationship investigations revealed that loss of the 3-(dichlorophenyl)-isoxazolyl moiety or the aromatic nitrogen heterocycle with nitrogen at α-position to the carbon of carboximidamide group significantly attenuated K18 antagonistic potency. Mutagenic studies supported by molecular dynamic simulations combined with Molecular Mechanics—Poisson Boltzmann Surface Area calculations identified the residues important for binding in the A3R orthosteric site. We demonstrate that K18, which contains a 3-(dichlorophenyl)-isoxazole group connected through carbonyloxycarboximidamide fragment with a 1,3-thiazole ring, is a specific A3R ( |
| Databáze: | OpenAIRE |
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