Compound profiling using a panel of steroid hormone receptor cell-based assays

Area of Science:

• Pharmacology
• Molecular Biology
• Drug Discovery

Background:

• Drug discovery for nuclear receptors (NRs) aims to enhance selectivity for reduced side effects.
• Selective modulation of NRs is key to targeting specific gene expression subsets.
• Cellular assays are crucial for evaluating functional activity, potency, and selectivity of NR modulators.

Purpose of the Study:

• To develop a comprehensive panel of cell-based assays for six key steroid hormone receptors.
• To enable high-throughput screening and selectivity profiling of potential drug candidates.
• To differentiate partial agonists, antagonists, and selective modulators.

Main Methods:

• Engineered stable cell lines expressing Gal4-fusion proteins for androgen, estrogen (alpha and beta), glucocorticoid, mineralocorticoid, and progesterone receptors.
• Validated assays using known agonists and antagonists for pharmacology and high-throughput compatibility.
• Profiled 35 compounds in dose-response across the assay panel in agonist and antagonist modes.

Main Results:

• Successfully developed and validated a panel of six steroid hormone receptor assays.
• Demonstrated the ability to identify selective estrogen receptor modulators (SERMs).
• Effectively detected mixed agonists/antagonists and partial agonists/antagonists.
• Chimeric assay data correlated well with literature data for full-length reporter gene assays.

Conclusions:

• The developed steroid hormone receptor assays offer high selectivity, sensitivity, and appropriate pharmacology.
• These assays are suitable for high-throughput screening and selectivity profiling of NR modulators.
• The panel facilitates the discovery of drugs with improved selectivity and reduced side effects.