Screening Assay for Inhibitors of TRPA1 Activation by a Lower Alkyl Phenol

A method for identifying a compound that modulates the activation of TRPA1 by Compound I, wherein Compound I is represented by Formula I:

• wherein each instance of R1 is independently hydrogen or a C1-6 alkyl, and n is an integer from 1-5, said method comprising:
• (a) incubating cells expressing TRPA1 with an indicator for a time sufficient to allow incorporation of the indicator into the cells, wherein the indicator is capable of signaling activation of TRPA1;
• (b) incubating said cells with a candidate compound while maintaining said cells in a solution that contains Compound I and is suitable to allow a flux of ions through TRPA1;
• (c) after completing steps (a) and (b), measuring a signal from the indicator in the presence of the candidate modulator compound;
• and
• (d) comparing the signal of step (c) to a standard value, wherein the indicator indicates the flux of ions via Compound I-activated TRPA1 and a difference between the signal of step (c) and the standard value indicates that the candidate compound modulates TRPA1 activation.

A method for identifying a compound that modulates the activation of TRPA1 by thymol, said method comprising:

• (a) incubating cells expressing TRPA1 with an indicator for a time sufficient to allow incorporation of the indicator into the cells, wherein the indicator is capable of signaling activation of TRPA1;
• (b) incubating said cells with a candidate compound while maintaining said cells in a solution that contains thymol and is suitable to allow a flux of ions through TRPA1;
• (c) after completing steps (a) and (b), measuring a signal from the indicator in the presence of the candidate modulator compound;
• and
• (d) comparing the signal of step (c) to a standard value, wherein the indicator indicates the flux of ions via thymol-activated TRPA1 and a difference between the signal of step (c) and the standard value indicates that the candidate compound modulates TRPA1 activation.