Agonists and Antagonists of Protease-Activated Receptor 2 Discovered within a DNA-Encoded Chemical Library Using Mutational Stabilization of the Target.

Autor:	Brown DG; 1 Discovery Sciences, Innovative Medicines and Early Development Biotech Unit, AstraZeneca R&D, Waltham, MA, USA., Brown GA; 2 Heptares Therapeutics Ltd., Welwyn Garden City, Hertfordshire, UK., Centrella P; 3 X-Chem Pharmaceuticals, Waltham, MA, USA., Certel K; 3 X-Chem Pharmaceuticals, Waltham, MA, USA.; 5 Novartis Institute for BioMedical Research, Cambridge, MA, USA., Cooke RM; 2 Heptares Therapeutics Ltd., Welwyn Garden City, Hertfordshire, UK., Cuozzo JW; 3 X-Chem Pharmaceuticals, Waltham, MA, USA., Dekker N; 4 Discovery Sciences, Innovative Medicines and Early Development Biotech Unit, AstraZeneca R&D, Mölndal, Sweden., Dumelin CE; 3 X-Chem Pharmaceuticals, Waltham, MA, USA.; 6 Novartis Institute for BioMedical Research, Basel, Switzerland., Ferguson A; 1 Discovery Sciences, Innovative Medicines and Early Development Biotech Unit, AstraZeneca R&D, Waltham, MA, USA.; 7 X-Chem, Inc., Waltham, MA, USA., Fiez-Vandal C; 2 Heptares Therapeutics Ltd., Welwyn Garden City, Hertfordshire, UK., Geschwindner S; 4 Discovery Sciences, Innovative Medicines and Early Development Biotech Unit, AstraZeneca R&D, Mölndal, Sweden., Guié MA; 3 X-Chem Pharmaceuticals, Waltham, MA, USA., Habeshian S; 3 X-Chem Pharmaceuticals, Waltham, MA, USA.; 8 École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland., Keefe AD; 3 X-Chem Pharmaceuticals, Waltham, MA, USA., Schlenker O; 2 Heptares Therapeutics Ltd., Welwyn Garden City, Hertfordshire, UK., Sigel EA; 3 X-Chem Pharmaceuticals, Waltham, MA, USA., Snijder A; 4 Discovery Sciences, Innovative Medicines and Early Development Biotech Unit, AstraZeneca R&D, Mölndal, Sweden., Soutter HT; 3 X-Chem Pharmaceuticals, Waltham, MA, USA., Sundström L; 4 Discovery Sciences, Innovative Medicines and Early Development Biotech Unit, AstraZeneca R&D, Mölndal, Sweden., Troast DM; 3 X-Chem Pharmaceuticals, Waltham, MA, USA.; 9 Morphic Therapeutic, Waltham, MA., Wiggin G; 2 Heptares Therapeutics Ltd., Welwyn Garden City, Hertfordshire, UK., Zhang J; 1 Discovery Sciences, Innovative Medicines and Early Development Biotech Unit, AstraZeneca R&D, Waltham, MA, USA.; 10 Entasis Therapeutics, Waltham, MA., Zhang Y; 3 X-Chem Pharmaceuticals, Waltham, MA, USA., Clark MA; 3 X-Chem Pharmaceuticals, Waltham, MA, USA.
Jazyk:	angličtina
Zdroj:	SLAS discovery : advancing life sciences R & D [SLAS Discov] 2018 Jun; Vol. 23 (5), pp. 429-436. Date of Electronic Publication: 2018 Jan 09.
DOI:	10.1177/2472555217749847
Abstrakt:	The discovery of ligands via affinity-mediated selection of DNA-encoded chemical libraries is driven by the quality and concentration of the protein target. G-protein-coupled receptors (GPCRs) and other membrane-bound targets can be difficult to isolate in their functional state and at high concentrations, and therefore have been challenging for affinity-mediated selection. Here, we report a successful selection campaign against protease-activated receptor 2 (PAR2). Using a thermo-stabilized mutant of PAR2, we conducted affinity selection using our >100-billion-compound DNA-encoded library. We observed a number of putative ligands enriched upon selection, and subsequent cellular profiling revealed these ligands to comprise both agonists and antagonists. The agonist series shared structural similarity with known agonists. The antagonists were shown to bind in a novel allosteric binding site on the PAR2 protein. This report serves to demonstrate that cell-free affinity selection against GPCRs can be achieved with mutant stabilized protein targets.
Databáze:	MEDLINE
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