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.
Department of Vascular Biology, Holland Laboratory, American Red Cross, Rockville, MD, USA. crandad@wyeth.com
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Crandall
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Elokdah
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Di
J K
JK
Hennan
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Gorlatova
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Lawrence
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