# Identification of Chemical Probes for the Orphan Nuclear Receptor NR2F6

> **NIH NIH R01** · SCRIPPS FLORIDA · 2020 · $469,049

## Abstract

PROJECT SUMMARY
Immune checkpoint therapy is proving to be an effective approach for the treatment of a variety of cancers.
However, only a subset of patients exhibits long-lasting responses highlighting the critical need for the
identification of novel options to augment effects. The nuclear receptor (NR) superfamily of ligand-regulated
transcription factors has proven to be an excellent source of targets for therapeutic intervention of a broad
range of diseases. The NR2F subfamily of NRs, COUP-TF1 (NR2F1), COUP-TFII (NR2F2), and COUP-TFIII
(NR2F6), are considered orphan receptors since their endogenous ligands have yet to be identified. NR2F6
has recently emerged as an intracellular T-cell immune checkpoint; NR2F6-deficient mice spontaneously reject
tumors and develop host-protective immunological memory. However, little is known about NR2F6's
transcriptional activity, which is due, in part, to lack of specific molecular probes that would enable interrogation
of its function. In order to identify ligands that modulate NR2F6 activity, we have designed a high-throughput
screening (HTS) compatible primary assay that specifically measures the transcriptional activity of NR2F6 in
vitro. Our goal is to implement a full HTS-campaign using the Scripps Institutional Drug Discovery Library
(SDDL) to identify, validate, and characterize potent small molecule modulators of NR2F6 activity. To achieve
our goal, we have miniaturized our assay into a 1,536-well plate format. Using LOPAC and a 10,000
compound library HTS screen we have validated our assay, meeting HTS automation criteria, and propose to
carry out a “full-deck” HTS campaign to screen the >640,000 compounds in the SDDL (Aim 1).
Cheminformatic analysis of “hits” will help identify the most promising leads by structural clustering,
bioinformatics analysis of compounds to determine promiscuity, and scaffold analysis to determine ease of
chemical synthesis and tractability for further medicinal chemistry efforts to perform structure-activity
relationship studies. In Aim 2, we will use a cascade of follow up assays to validate screening hits, determine
specificity, and begin to understand mechanism of action of NR2F6-mediated transcriptional activity. Finally,
in Aim 3, validated HTS hits will be advanced to early medicinal chemistry for lead optimization and
characterization of novel NR2F6 molecular probes. We expect that completion of this application will deliver
multiple structurally distinct NR2F6 modulators that exhibit suitable levels of cellular activity, potency, and
selectivity. Further medicinal chemistry efforts will enable development of lead compounds to determine
potency and specificity for NR2F6 while minimizing toxicity. Our collaborative research team has a strong
track record of performing high-throughput screens, selective optimization of scaffolds, and in vitro and in vivo
characterization of compounds. Collectively, our screening approach puts us in a unique position to identify,...

## Key facts

- **NIH application ID:** 9928414
- **Project number:** 5R01CA225890-03
- **Recipient organization:** SCRIPPS FLORIDA
- **Principal Investigator:** Louis Daniel Scampavia
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $469,049
- **Award type:** 5
- **Project period:** 2018-06-15 → 2022-04-01

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9928414

## Citation

> US National Institutes of Health, RePORTER application 9928414, Identification of Chemical Probes for the Orphan Nuclear Receptor NR2F6 (5R01CA225890-03). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/9928414. Licensed CC0.

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