Fig. 2: AZ2429 binds to PAR2 and activates multiple signalling pathways.
a Chemical structure of most active enantiomer of the potent agonist compound (AZ2429). AZ2429 (blue) and known PAR2 agonists SLIGKV-NH2 (black) and GB110 (grey) were pharmacologically profiled for: b competitive binding against [3H]-GB110 in membrane preparations of HEKexpi293 cells overexpressing human PAR2; c Ca2+ mobilisation and d IP1 production in 1321N1-hPAR2 cells; as well as e β-arrestin-2 recruitment and f phosphorylation of ERK1/2 in U2OS-hPAR2 cells. No agonistic effects were detected in parental 1321N1 cells, exemplified in the Ca2+ assay (open symbols). Agonist responses were calculated as % maximum response of SLIGKV-NH2. Graphs show representative data of 2 or more experiments, presented as individual data points with error bars denoting the s.e.m. The comprehensive pharmacological data are stated in Table 2. g Refined model of AZ2429 (blue) in the PAR2 receptor generated through molecular docking calculations in an adapted model of the PAR2 structure. Superimposed is AZ8838 (magenta) from the crystal structure (PDB 5NDD) showing the overlap of the benzyl substituents of each compound. Important receptor side chains are colour-coordinated based on their spatial localisation.
