A Robust and Efficient FRET-Based Assay for Cannabinoid Receptor Ligands Discovery

Area of Science:

• Pharmacology
• Biochemistry
• Drug Discovery

Background:

• Cannabinoid Receptors (CBRs) are crucial drug targets due to their involvement in various pathophysiological conditions.
• Developing sensitive high-throughput screening (HTS) assays is vital for identifying novel CBR modulators in drug discovery.
• Existing screening methods require optimization for efficiency and reliability in hit identification.

Purpose of the Study:

• To develop and validate a novel Tag-lite® binding assay for screening human Cannabinoid Receptor 1 (hCB1) and Cannabinoid Receptor 2 (hCB2) binding.
• To assess the assay's suitability for hit identification campaigns in drug discovery.

Main Methods:

• Development of a Tag-lite® binding assay utilizing a dual fluorescent ligand, CELT-335.
• Validation of the assay using known CBR ligands with diverse affinity and functional profiles.
• Implementation of a homogeneous assay format for enhanced screening efficiency.

Main Results:

• The Tag-lite® assay demonstrated robustness and high sensitivity for screening hCB1/hCB2 binding.
• CELT-335 proved to be an effective fluorescent ligand, ensuring optimal assay performance.
• The assay successfully differentiated between compounds with varying affinities and functional profiles.

Conclusions:

• The developed Tag-lite® assay is a reliable and sensitive method for screening CBR modulators.
• This assay is well-suited for hit and lead identification campaigns in drug discovery targeting Cannabinoid Receptors.
• The homogeneous format and fluorescent ligand contribute to the assay's efficiency and viability.