Unprecedented Fragment-Based screening using Spectral Shift for GPCRs

Описание: Specifically, we will outline how our experts and internal capabilities can speed up your drug discovery processes taking advantage of their expertise in GPCRs protein production and screening with the most recent biophysics technologies.
Fragment-based drug design presents a unique opportunity to identify novel binding sites on membrane proteins. We leveraged fragment screening using the most recent biophysical technology, Spectral Shift, to discover antagonist hit fragments on the human adenosine A2A receptor. This receptor belongs to the G-protein-coupled receptor (GPCR) family, expressed in the plasma membrane of neurons and immune cells, including macrophages, cytotoxic and regulatory T cells. Its abnormal expression in immune cells inhibits the immune response and promotes the growth and escape of tumor cells from immune cells. Eurofins CALIXAR optimized the production of the wild-type, full-length, and active recombinant protein solubilized from Sf9 insect cells using Eurofins CALIXAR proprietary detergents.
We will highlight the advantages of Spectral Shift technology, one of the possible approaches using the Dianthus (well-known for the MST-TRIC). Using Echo® acoustic droplet ejection, and specifically nanoliter dispensing in 384-well plates, robust data by Spectral Shift were generated for the fragment-based screening campaign with a 9.2% hit rate. Among 86 fragments, 20 fragment hits were confirmed by dose-response testing, with µM-range affinity values. NanoDSF was then used as an orthogonal assay for the identification of stabilizers. Additionally, the fragment hits were tested with one of our cell based LeadHunter® assays (A2A receptor), and interestingly, some were found to be antagonists at high concentration with very good ligand efficiency. Docking studies to identify the fragment binding sites are currently ongoing. The starting points will be used as the basis for fragment growing.