Platelet GPIIb/IIIa receptor occupancy studies using a novel fluoresceinated cyclic Arg-Gly-Asp peptide.
DMP 728 is a potent and specific platelet GPIIb/IIIa antagonist. Like all GPIIb/IIIa antagonists, DMP 728 has a steep dose-response relationship in inhibiting platelet aggregation. In this study the relationships between receptor occupancy, platelet aggregation and bleeding time was determined in anesthetized dogs after intravenous infusion of DMP 728 (0.01 and 0.1 mg/kg/2h). Receptor occupancy was determined by flow cytometry using XL086, a novel fluorescent cyclic RGD peptide that binds to GPIIb/IIIa with high specificity and affinity (kd approximately 55 nM). Mean number of GPIIb/IIIa as determined by flow cytometric assay was approximately 53,8000 and 79,000 on unactivated and ADP-activated platelets respectively. After DMP 728 intravenous infusion, there was a dose- and time-dependent increase in receptor occupancy, inhibition of platelet aggregation and bleeding time. The two methods of receptor occupancy determination correlate with each other with an r2 = 0.78. The present data suggest that blockade of only 40-60% (approximately 40,000 receptors) of the total platelet GPIIb/IIIa was required to achieve > 90% inhibition of platelet aggregation and > 15 min bleeding time. Our results showed the potential clinical utility of this approach in the study of GPIIb/IIIa dose-response relationship.
Tsao
P W
PW
DuPont Merck Pharmaceutical Company, Cardiovascular Diseases Division, Wilmington, DE 19880-0400, USA.
Bozarth
J M
JM
Jackson
S A
SA
Forsythe
M S
MS
Flint
S K
SK
Mousa
S A
SA
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D002138:Calibration; D004285:Dogs; D019793:Fluorescein; D005452:Fluoresceins; D006801:Humans; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000097:blood*; D010456:Peptides, Cyclic / Q000097:blood*; D010980:Platelet Membrane Glycoproteins / Q000037:antagonists & inhibitors*; D012044:Regression Analysis
D005452:Fluoresceins; D009842:Oligopeptides; D010456:Peptides, Cyclic; D010980:Platelet Membrane Glycoproteins; C047981:arginyl-glycyl-aspartic acid; D019793:Fluorescein