Oral Antithrombotic Effects of Acylhydrazone Derivatives

Abstract

Aim:In the search for new antithrombotic drug candidates, the synthesis and anti-platelet activity of a new series of N-acylhydrazones that were designed as thrombin inhibitors has been previously described. The aim of this work was to further characterize the effects of these compounds on thrombin-induced platelet aggregation and induced thrombosis in vivo. Methods: In this work, four compounds were tested, LASSBio-693, 694, 743 and 752, on platelet aggregation induced by thrombin, ADP and TRAP-4A. These compounds were further tested using a mouse pulmonary thromboembolism model induced by collagen (500 μg/kg) and norepinephrine (80 μg/kg) or thrombin (2,000 UI), and a deep venous thrombosis model.

New approaches in tail‐bleeding assay in mice: improving an important method for designing new anti‐thrombotic agents

“…; Song & Wang ; Frattani et al . ). Dejana and co‐workers compared these two techniques and showed that the bleeding time is significantly longer in the transection model (Dejana et al .…”

“…; Song & Wang ; Frattani et al . ). In the filter paper method, the operator may compromise the assays during blotting by pushing the injury against the paper and traumatizing the tail, wrongly inducing longer bleeding times (Greene et al .…”

“…; Song & Wang ; Frattani et al . ). However, the temperature of the solution may interfere in the assay as bleeding times detected at 37°C can be significantly shortened due to the higher blood flow in comparison with a room temperature counterpart (Dejana et al .…”

“…; Song & Wang ; Frattani et al . ), (ii) sharpness of the scalpel and/or surgical knife inducing crush injury and (iii) instrument used (razor blade, scalpel, surgical knife, retractable knife) (Greene et al . ).…”

“…Currently, bleeding induction assays in mice involve (i) a small incision (template model) (Stella et al 1975;Dejana et al 1979;Chelucci et al 2014) or (ii) total amputation of the tip of the tail (transection model) (Dejana et al 1979;Herbert & Bernat 1993;Tanaka et al 1998;Elg et al 1999;Gadi et al 2009;Greene et al 2010;Liu et al 2012;Song & Wang 2012;Frattani et al 2013). Dejana and coworkers compared these two techniques and showed that the bleeding time is significantly longer in the transection model (Dejana et al 1979).…”