High Throughput Screening Assay for the TRPM5 Ion Channel
AssigneeRedpoint Bio Corporation
InventorsRobert W. BRYANT, S. Paul Lee, R. Kyle Palmer, Qifeng Yang, M.N. Tulu Buber
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Abstract
There exists a need in the art for high throughput screening assays that can identify compounds that specifically modulate the activity of fast-acting ion channels, such as TRPM5. Current methods suffer from a lack of sensitivity, low throughput, and are labor intensive. The claimed methods provide fluorescent assays with an optical readout that gives rapid readout of the results, has a high signal to noise background ratio, are easy to use, can be modified for automation and miniaturization, and provide verification that a compound specifically modulates TRPM5.
Claims23
1-14. (canceled)
A high throughput screening assay for determining whether a test compound is a TRPM5 ion channel-specific modulator comprising:
- (a) contacting a cell that expresses TRPM5 and has been preloaded with a membrane potential fluorescent dye, with a test compound in the presence of potassium chloride;
- (b) using an optical detector, measuring the fluorescent intensity of said cell in the presence of said potential modulating compound;
- (c) comparing the measured fluorescent intensity determined in step (b) to the fluorescent intensity of a different cell that expresses TRPM5 and has been preloaded with a membrane potential fluorescent dye, in the presence of potassium chloride and the absence of the test compound;
- and
- (d) evaluating whether the test compound may be a TRPM5-specific modulator by determining if the ratio of the fluorescent intensity with potassium chloride and the test compound to the intensity with potassium chloride in the absence of the test compound is less than or greater than 1.
23-26. (canceled)
30-63. (canceled)
Dependencies are inferred from the claim text.