Journal
JOURNAL OF THROMBOSIS AND HAEMOSTASIS
Volume 10, Issue 5, Pages 870-880Publisher
WILEY
DOI: 10.1111/j.1538-7836.2012.04679.x
Keywords
anticoagulant; antidote; aptamer; exosite I; prothrombin; thrombin
Categories
Funding
- American Heart Association [10PRE3260011]
- NIH [R01HL65222]
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. Background:similar to The conversion of prothrombin to thrombin is one of two non-duplicated enzymatic reactions during coagulation. Thrombin has long been considered an optimal anticoagulant target because it plays a crucial role in fibrin clot formation by catalyzing the cleavage of fibrinogen, upstream coagulation cofactors and platelet receptors. Although a number of anti-thrombin therapeutics exist, it is challenging to use them clinically due to their propensity to induce bleeding. Previously, we isolated a modified RNA aptamer (R9D-14) that binds prothrombin with high affinity and is a potent anticoagulant in vitro. Objectives: We sought to explore the structure of R9D-14 and elucidate its anticoagulant mechanism(s). In addition to designing an optimized aptamer (RNAR9D-14T), we also explored whether complementary antidote oligonucleotides can rapidly modulate the optimized aptamers anticoagulant activity. Methods and Results:similar to RNAR9D-14T binds prothrombin and thrombin pro/exosite I with high affinity and inhibits both thrombin generation and thrombin exosite I-mediated activity (i.e. fibrin clot formation, feedback activity and platelet activation). RNAR9D-14T significantly prolongs the aPTT, PT and TCT clotting assays, and is a more potent inhibitor than the thrombin exosite I DNA aptamer ARC-183. Moreover, a complementary oligonucleotide antidote can rapidly (< 2 min) and durably (>2 h) reverse RNAR9D-14T anticoagulation in vitro. Conclusions:similar to Powerful anticoagulation, in conjunction with antidote reversibility, suggests that RNAR9D-14T may be ideal for clinical anticoagulation in settings that require rapid and robust anticoagulation, such as cardiopulmonary bypass, deep vein thrombosis, stroke or percutaneous coronary intervention.
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