Identification of positive modulators of TRPM5 channel from a high-throughput screen using a fluorescent membrane
Caterina Virginio1, Laura Aldegheri1, Selena Nola1
1Aptuit, an Evotec Company, Via Alessandro Fleming, 4, 37135 Verona, Italy.
Abstract:
Transient Receptor Potential Melastatin 5 (TRPM5) is an intracellular calcium-activated cation-selective ion channel expressed in a variety of cell types. Dysfunction of this channel has recently been implied in a range of disease states including diabetes, enteric infections, inflammatory responses, parasitic infection and other pathologies. However, to date, agonists and positive modulators of this channel with sufficient selectivity to enable target validation studies have not been described, limiting the evaluation of TRPM5 biology and its potential as a drug target. We developed a high-throughput assay using a fluorescent membrane potential dye and a medium- and high-throughput electrophysiology assay using QPatch HTX and SyncroPatch 384PE. By employing these assays, we conducted a primary screening campaign and identified hit compounds as TRPM5 channel positive modulators. An initial selectivity profile confirmed hit selectivity to TRPM5 and is presented here. These small molecule TRPM5 compounds have a high potential both as early tool compounds to enable pharmacological studies of TRPM5 and as starting points for the development of potent, selective TRPM5 openers or positive modulators as novel drugs targeting several pathological states.
Insights
Researchers identified novel small molecules that activate the Transient Receptor Potential Melastatin 5 (TRPM5) channel. These compounds are valuable tools for studying TRPM5 in diseases like diabetes and inflammation.
Area of Science:
- Ion channel pharmacology
- Molecular and cellular biology
Background:
- Transient Receptor Potential Melastatin 5 (TRPM5) is a calcium-activated ion channel implicated in various diseases.
- Lack of selective TRPM5 agonists/modulators hinders research and drug development.
Purpose of the Study:
- To develop and utilize high-throughput screening assays for identifying TRPM5 positive modulators.
- To discover novel small molecules for TRPM5 target validation and therapeutic development.
Main Methods:
- Development of fluorescent membrane potential and electrophysiology assays (QPatch HTX, SyncroPatch 384PE).
- High-throughput screening campaign to identify hit compounds.
- Initial selectivity profiling of identified compounds.
Main Results:
- Successful identification of hit compounds acting as TRPM5 channel positive modulators.
- Confirmation of TRPM5 selectivity for the identified compounds.
- Demonstration of compounds' potential as pharmacological tools.
Conclusions:
- Novel small molecule TRPM5 positive modulators have been discovered.
- These compounds serve as crucial tools for TRPM5 research and potential drug leads for diseases involving TRPM5 dysfunction.
