Synthetic biotinylated peptide compounds derived from Asp-hemolysin: Novel potent inhibitors of platelet-activating factor

Platelet-activating factor (PAF: 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine), a potent inflammatory mediator, is implicated in many inflammatory diseases and may possibly serve as a direct target for anti-inflammatory drugs. We have previously reported that Asp-hemolysin-related synthetic peptides (P4−P29) inhibit the bioactivities of oxidized low-density lipoprotein (ox-LDL) containing PAF-like lipids by direct binding to ox-LDL, which plays a key role in the atherosclerotic inflammatory process. In this study, we investigated whether these peptides inhibit the bioactivities of PAF by binding to PAF and its metabolite/precursor lyso-PAF. In in vitro experiments, P21, one of the peptides, bound to both PAF and lyso-PAF in a dose-dependent manner and markedly inhibited PAF-induced apoptosis in human umbilical vein endothelial cells. Moreover, in in vivo experiments, P4 and P21, particularly their N-terminally biotinylated peptide compounds (BP4 and BP21), inhibited PAF-induced rat paw oedema dose dependently and markedly, and showed sufficient inhibition of the oedema even at doses 150–300 times less than the doses of PAF antagonists. These results provide evidence that direct binding of N-terminally biotinylated peptide compounds derived from Asp-hemolysin to PAF and lyso-PAF leads to a dramatic inhibition of the bioactivities of PAF, both in vitro and in vivo, and strongly suggesting that these compounds may be useful as a novel type of anti-inflammatory drug for the treatment of several inflammatory diseases caused by PAF.