A new method for measurement of plasma concentration of orally active glycoprotein IIb/IIIa antagonists.
A bioassay for determining concentrations of antiplatelet compounds in plasma or aqueous solution has been developed. The method uses an aliquot of plasma from treated animals to inhibit collagen-induced platelet aggregation in pooled platelet-rich plasma (PRP) obtained from donor dogs. The concentration in plasma from treated animals was estimated using a standard curve of inhibition established using plasma from untreated animals which had been spiked with known amounts of compound. For independent validation, plasma concentrations of certain compounds were determined in identical dog plasma samples by both bioassay and HPLC. Results from the two methods were concordant. The bioassay provides an accurate and sensitive method for measuring antiplatelet activity without the need for extraction of plasma samples and may be used to measure activity in any solution which is compatible with PRP. This assay is routinely used to provide an estimate of absorption of prodrugs and systemic conversion to active compound after oral dosing. Some of the compounds of interest are ester-acid pairs with the inactive ester prodrug being cleaved to the active acid following administration. Compounds were administered orally (ester) or IV (acid) and blood samples were taken periodically for 24 hours. Plasma concentration of active moiety was determined for each time point and the area under the curve (AUC) of concentration vs. time was calculated. Comparing the AUCs for oral and IV routes of administration yielded the Oral Systemic Activity (OSA), a measure of active compound available after oral dosing.
Salyers
A K
AK
Szalony
J A
JA
Taite
B B
BB
Haas
N F
NF
Mehrotra
D V
DV
Feigen
L P
LP
Nicholson
N S
NS
D000284:Administration, Oral; D000818:Animals; D001681:Biological Assay; D018592:Cross-Over Studies; D004285:Dogs; D010975:Platelet Aggregation Inhibitors / Q000097:blood*; D010980:Platelet Membrane Glycoproteins / Q000037:antagonists & inhibitors*; D011897:Random Allocation; D015203:Reproducibility of Results
D010975:Platelet Aggregation Inhibitors; D010980:Platelet Membrane Glycoproteins