Yoda1 analogue (<scp>D</scp>ooku1) which antagonizes <scp>Y</scp>oda1‐evoked activation of <scp>P</scp>iezo1 and aortic relaxation

TLDR

The mechanosensitive Piezo1 channel plays key roles in vascular physiology and disease. The authors synthesized a Yoda1 analogue, Dooku1, that lacks agonist activity but reversibly antagonizes Yoda1, and tested its effects using Ca²⁺ and Tl⁺ imaging in HEK293/CHO/HUVEC cells and isometric tension recordings in mouse aortic rings. Dooku1 potently blocks Yoda1‑induced Piezo1 activation (IC₅₀ ≈1.3–1.5 µM) and Yoda1‑mediated aortic relaxation, without affecting constitutive Piezo1 activity or other Ca²⁺ pathways, indicating a specific chemical antagonism site.

Abstract

The mechanosensitive Piezo1 channel has important roles in vascular physiology and disease. Yoda1 is a small-molecule agonist, but the pharmacology of these channels is otherwise limited.Yoda1 analogues were generated by synthetic chemistry. Intracellular Ca2+ and Tl+ measurements were made in HEK 293 or CHO cell lines overexpressing channel subunits and in HUVECs, which natively express Piezo1. Isometric tension recordings were made from rings of mouse thoracic aorta.Modification of the pyrazine ring of Yoda1 yielded an analogue, which lacked agonist activity but reversibly antagonized Yoda1. The analogue is referred to as Dooku1. Dooku1 inhibited 2 μM Yoda1-induced Ca2+ -entry with IC50 s of 1.3 μM (HEK 293 cells) and 1.5 μM (HUVECs) yet failed to inhibit constitutive Piezo1 channel activity. It had no effect on endogenous ATP-evoked Ca2+ elevation or store-operated Ca2+ entry in HEK 293 cells or Ca2+ entry through TRPV4 or TRPC4 channels overexpressed in CHO and HEK 293 cells. Yoda1 caused dose-dependent relaxation of aortic rings, which was mediated by an endothelium- and NO-dependent mechanism and which was antagonized by Dooku1 and analogues of Dooku1.Chemical antagonism of Yoda1-evoked Piezo1 channel activity is possible, and the existence of a specific chemical interaction site is suggested with distinct binding and efficacy domains.