Discovery of RORγ Allosteric Fluorescent Probes and Their Application: Fluorescence Polarization, Screening, and...

Area of Science:

• Biochemistry and Molecular Biology
• Immunology
• Medicinal Chemistry

Background:

• Retinoic acid receptor-related orphan receptor gamma (RORγ) is a key transcription factor in Th17 cells, implicated in various autoimmune conditions.
• RORγ allosteric inhibitors represent a promising therapeutic strategy to modulate RORγ transcriptional activity.

Purpose of the Study:

• To design, synthesize, and characterize novel RORγ allosteric fluorescent probes.
• To establish a fluorescence polarization-based assay for screening RORγ allosteric binders.
• To identify new RORγ allosteric inhibitors using virtual screening and the developed assay.

Main Methods:

• Design and synthesis of 11 RORγ allosteric fluorescent probes based on MRL-871.
• Development of a fluorescence polarization assay using probe 12h.
• Virtual screening combined with the developed assay to identify novel inhibitors.

Main Results:

• Successful synthesis and characterization of 11 allosteric fluorescent probes.
• Establishment of an efficient and cost-effective fluorescence polarization assay.
• Identification of 10 novel RORγ allosteric inhibitors, including hit compound G381-0087.
• Initial structure-activity relationship (SAR) studies on G381-0087.

Conclusions:

• Probe 12h is a valuable tool for RORγ allosteric inhibitor discovery.
• The developed screening platform facilitates the exploration of RORγ allosteric inhibitors.
• This work contributes to a better understanding of RORγ function in autoimmune disorders.