Improved PIEZO1 agonism through 4-benzoic acid modification of Yoda1

Background and Purpose

PIEZO1 forms mechanically activated calcium-permeable non-selective cation channels in numerous species and cell types. Options for pharmacological modulation are limited and so we modified a PIEZO1 small-molecule agonist (Yoda1) to advance capability for modulation.

Experimental Approach

Medicinal chemistry generated Yoda1 analogues that were tested in intracellular calcium and patch-clamp assays on cultured cells exogenously expressing human or mouse PIEZO1 or mouse PIEZO2, physico-chemical assays and wire myography assays on vein from mice in which there was genetic disruption of PIEZO1.

Key Results

A Yoda1 analogue (KC159) containing 4-benzoic acid instead of the pyrazine of Yoda1 and its potassium salt (KC289) have equivalent or improved reliability, efficacy and potency compared to Yoda1 in functional assays. Tested against over-expressed mouse PIEZO1 in calcium assays, potency order is KC289 (150)>KC159 (280)>Yoda1 (600) (EC50s in parentheses, nM). There is selectivity for PIEZO1 over other membrane proteins and the physico-chemical properties are more suited to physiological conditions than those of Yoda1. Vasorelaxant effects occur that are consistent with PIEZO1 agonism. 2-benzoic acid substitution, by contrast, fails to generate a modulator.

Conclusion and Implications

4-Benzoic acid modification of Yoda1 improves PIEZO1 agonism. We suggest naming this new modulator Yoda2. It should be a useful tool compound in physiological assays and facilitate efforts to identify a binding site. Such compounds may have therapeutic potential, for example in diseases linked genetically to PIEZO1 such as lymphatic dysplasia.