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Yoda1 analogue ( D ooku1) which antagonizes Y oda1‐evoked activation of P iezo1 and aortic relaxation
Author(s) -
Evans Elizabeth L,
Cuthbertson Kevin,
Endesh Naima,
Rode Baptiste,
Blythe Nicola M,
Hyman Adam J,
Hall Sally J,
Gaunt Hannah J,
Ludlow Melanie J,
Foster Richard,
Beech David J
Publication year - 2018
Publication title -
british journal of pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.432
H-Index - 211
eISSN - 1476-5381
pISSN - 0007-1188
DOI - 10.1111/bph.14188
Subject(s) - hek 293 cells , piezo1 , mechanosensitive channels , chemistry , agonist , trpv4 , microbiology and biotechnology , intracellular , biophysics , ion channel , biochemistry , receptor , biology
Background and Purpose 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. Experimental Approach Yoda1 analogues were generated by synthetic chemistry. Intracellular Ca 2+ 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. Key Results 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 Ca 2+ ‐entry with IC 50 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 Ca 2+ elevation or store‐operated Ca 2+ entry in HEK 293 cells or Ca 2+ 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. Conclusion and Implications 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.