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Updated: Jan 2, 2026

Characterization of G Protein-coupled Receptors by a Fluorescence-based Calcium Mobilization Assay
Published on: July 28, 2014
Ligand-Based Design of Allosteric Retinoic Acid Receptor-Related Orphan Receptor γt (RORγt) Inverse Agonists
Femke A Meijer1, Richard G Doveston1,2, Rens M J M de Vries1
1Laboratory of Chemical Biology, Department of Biomedical Engineering and Institute for Complex Molecular Systems , Technische Universiteit Eindhoven , Den Dolech 2 , 5612 AZ Eindhoven , The Netherlands.
Researchers discovered novel allosteric inverse agonists targeting retinoic acid receptor-related orphan receptor gamma t (RORγt) using an in silico approach. These compounds effectively inhibit RORγt activity, offering a new strategy for autoimmune disease treatment.
Area of Science:
- Medicinal Chemistry
- Molecular Biology
- Pharmacology
Background:
- Retinoic acid receptor-related orphan receptor gamma t (RORγt) is implicated in autoimmune disease pathogenesis.
- Allosteric inhibition presents a novel and advantageous therapeutic strategy for RORγt compared to orthosteric methods.
Purpose of the Study:
- To discover novel allosteric RORγt inverse agonists using an efficient in silico-guided approach.
- To characterize the identified compounds for their inhibitory potential and mechanism of action.
Main Methods:
- In silico screening and ligand-based design to identify novel chemical scaffolds.
- Biochemical assays (coactivator recruitment) to assess RORγt inhibition.
- Cell-based assays to measure IL-17a mRNA production.
- Cocrystallization studies to confirm the allosteric binding mode.
Main Results:
- Discovery of a novel isoxazole chemotype with potent allosteric inverse agonism against RORγt.
- Compound 25 (FM26) demonstrated submicromolar inhibition and effectively reduced IL-17a mRNA levels.
- Biochemical and crystallographic data confirmed the allosteric mechanism of action for compound 25.
- Isoxazole compounds exhibited promising pharmacokinetic properties and chemical diversity.
Conclusions:
- The study successfully identified novel allosteric RORγt inverse agonists via an efficient in silico approach.
- The discovered isoxazole compounds represent a promising new class of therapeutics for RORγt-mediated autoimmune diseases.
- The employed ligand-based design strategy is versatile for targeting allosteric sites on RORγt.
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