Characterization and comparative evaluation of a structurally unique PAI-1 inhibitor exhibiting oral in-vivo efficacy.
Plasminogen activator inhibitor-1 (PAI-1) is the major physiological inhibitor of both tissue-type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA). Elevated levels of PAI-1 are associated with thrombosis and vascular disease, suggesting that high plasma PAI-1 may promote a hypercoagulable state by disrupting the natural balance between fibrinolysis and coagulation. In this study, we identify WAY-140312 as a structurally novel small molecule inactivator of PAI-1, compare its inhibitory activity with other previously identified small molecule inhibitors, and investigate the mechanism of inactivation of PAI-1 in the presence of both tPA and uPA. In an immunofunctional assay, WAY-140312 inhibited PAI-1 with an estimated inhibitory concentration (IC(50)) of 11.7 micro m, which was the lowest value obtained of the four different PAI-1 inactivators tested. Surface activity profiling indicated that the critical micelle concentration for WAY-140312 was 95.8 micro m, and that each inhibitor exhibited unique physical chemical properties. Using a sensitive direct activity assay, the IC(50) for WAY-140312 was similar when either tPA or uPA was used as the target protease. Immunoblot analysis demonstrated that WAY-140312 near the IC(50) inhibited the complex formation between either tPA or uPA and PAI-1. After oral administration, WAY-140312 exhibited 29% bioavailability with a plasma half-life of approximately 1 h. In an in-vivo model of vascular injury, a 10 mg kg(-1) oral dose of WAY-140312 was associated with improvement in arterial blood flow and reduction in venous thrombosis. Thus, WAY-140312 represents a structurally novel small molecule inhibitor of PAI-1, and is the first such molecule to exhibit efficacy in animal models of vascular disease following oral administration.
Crandall
D L
DL
Department of Vascular Biology, Holland Laboratory, American Red Cross, Rockville, MD, USA. crandad@wyeth.com
Elokdah
H
H
Di
L
L
Hennan
J K
JK
Gorlatova
N V
NV
Lawrence
D A
DA
D000284:Administration, Oral; D000818:Animals; D001158:Arteries / Q000473:pathology; D001572:Benzofurans / Q000737:chemistry* / Q000494:pharmacology*; D001774:Blood Chemical Analysis / Q000379:methods*; D001777:Blood Coagulation; D002339:Carotid Arteries / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D005342:Fibrinolysis; D007118:Immunoassay; D015151:Immunoblotting; D020128:Inhibitory Concentration 50; D008823:Micelles; D008956:Models, Chemical; D017395:Plasminogen Activator Inhibitor 1 / Q000737:chemistry*; D015849:Plasminogen Inactivators / Q000097:blood / Q000494:pharmacology*; D051381:Rats; D013777:Tetrazoles / Q000737:chemistry* / Q000494:pharmacology*; D013927:Thrombosis; D013997:Time Factors; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism
D001572:Benzofurans; D008823:Micelles; D017395:Plasminogen Activator Inhibitor 1; D015849:Plasminogen Inactivators; D013777:Tetrazoles; C496986:WAY-140312; D014568:Urokinase-Type Plasminogen Activator