Molecular aspects of cloricromene (AD6) distribution in human platelets and its pharmacological effects.

Cloricromene (AD6) is an investigational drug which inhibits platelet aggregation and release reaction. We studied the relationship between its action and its distribution and metabolism in platelets. Incubation of anticoagulated whole blood or platelet-rich plasma (PRP) without exogenous aggregating agents resulted in a progressive decrease of platelet count with a concomitant increase of beta-thromboglobulin (BTG) release. AD6 (20-50 mumols/l), but not acetylsalicylic acid (ASA), incubated with whole blood or PRP, prevented the fall in platelet count and the release of BTG for at least 150 min. Moreover, incubation of PRP with AD6 (50 mumols/l) and subsequent stimulation by ADP at threshold concentrations resulted in a significant reduction (about 30%) in aggregation for at least 90 min. AD6 (20 mumols/l) added to PRP was rapidly metabolized by hydrolysis of an ester bond to AD6 acid, a stable catabolite pharmacologically inactive in platelets. Significant amounts of AD6 acid (up to 13.26 +/- 2.80 pmol/10(6) platelets) were associated with the platelets after incubation either at 37 degrees C or 4 degrees C. The amount of AD6 acid in the platelet pellet was proportional to AD6 concentration (2 to 100 mumols/l). PRP incubation with AD6 acid (20 mumols/l) resulted in very low levels (less than 1 pmol/10(6) platelets) of the same compound in the platelet pellet after 1, 5 or 30 min. These data suggest that AD6 is taken up as an ester and converted to its acid catabolite with a consequent long-lasting inhibition of platelet function.