Development of a Testing Funnel for Identification of Small-Molecule Modulators Targeting Secretin Receptors

Dengler DG, Sun Q, Holleran J, Pollari S, Beutel J, Brown BT, Iwaya AS, Ardecky R, Harikumar KG, Miller LJ, Sergienko EA (2020)

Publication Type: Journal article

Publication year: 2020

Journal

SLAS Discovery SAGE Publications

DOI: 10.1177/2472555220945284

Abstract

The secretin receptor (SCTR), a prototypical class B G protein-coupled receptor (GPCR), exerts its effects mainly by activating Gαs proteins upon binding of its endogenous peptide ligand secretin. SCTRs can be found in a variety of tissues and organs across species, including the pancreas, stomach, liver, heart, lung, colon, kidney, and brain. Beyond that, modulation of SCTR-mediated signaling has therapeutic potential for the treatment of multiple diseases, such as heart failure, obesity, and diabetes. However, no ligands other than secretin and its peptide analogs have been described to regulate SCTRs, probably due to inherent challenges in family B GPCR drug discovery. Here we report creation of a testing funnel that allowed targeted detection of SCTR small-molecule activators. Pursuing the strategy to identify positive allosteric modulators (PAMs), we established a unique primary screening assay employing a mixture of three orthosteric stimulators that was compared in a screening campaign testing 12,000 small-molecule compounds. Beyond that, we developed a comprehensive set of secondary assays, such as a radiolabel-free target engagement assay and a NanoBiT (NanoLuc Binary Technology)-based approach to detect β-arrestin-2 recruitment, all feasible in a high-throughput environment as well as capable of profiling ligands and hits regarding their effect on binding and receptor function. This combination of methods enabled the discovery of five promising scaffolds, four of which have been validated and further characterized with respect to their allosteric activities. We propose that our results may serve as starting points for developing the first in vivo active small molecules targeting SCTRs.

Authors with CRIS profile

Jannis Beutel Lehrstuhl für Pharmazeutische Chemie

Involved external institutions

Sanford Burnham Prebys Medical Discovery Institute US United States (USA) (US) Mayo Clinic US United States (USA) (US)

How to cite

APA:

Dengler, D.G., Sun, Q., Holleran, J., Pollari, S., Beutel, J., Brown, B.T.,... Sergienko, E.A. (2020). Development of a Testing Funnel for Identification of Small-Molecule Modulators Targeting Secretin Receptors. SLAS Discovery. https://dx.doi.org/10.1177/2472555220945284

MLA:

Dengler, Daniela G., et al. "Development of a Testing Funnel for Identification of Small-Molecule Modulators Targeting Secretin Receptors." SLAS Discovery (2020).

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