Figure 4. 3D53 is more effective than W54011 and JJ47 in inhibiting C5a-induced migration or calcium release for human HMDM due to longer residence time on C5aR.

(A) Migration of HMDM through 5 μm Transwell inserts with or without 3 nM C5a for 16 h. For antagonism of C5aR, cells were treated with 3D53 (0.1 or 1 μM), W54011 (0.1 or 1 μM) or JJ47 (0.1 or 1 μM) for 1 h. Unbound antagonists were washed away prior to stimulation with 3 nM C5a. Chemotactic index is expressed as fold change against control. (B) C5a induced Ca²⁺ release from HMDM, pre-treated with 1 μM C5aR antagonist (●) 3D53, (▲) W54011 and (○) JJ47 for 1 h. Excess unbound antagonists were removed by washing. Antagonist residence time on C5aR was determined by subjecting treated cells to 3 nM C5a during the stated duration post antagonist-treatment up to 4 h and (C) up to 60 h for 3D53 in a calcium release assay. Calculated half-life for 3D53, W54011 and JJ47 are 18.2 h, 1.2 h and 0.6 h respectively. Error bars are means ± SEM of three independent experiments (n = 3). ***p < 0.001 by student t-test.