title: In Vitro Evaluation of Aptamer-Based Reversible Inhibition of Anticoagulant Activated Protein C as a Novel Supportive Hemostatic Approach.
abstract: Activated protein C (APC) is a critical regulator of thrombin formation and thereby protects against thrombosis. On the other hand, overwhelming formation of APC increases the risk of bleeding such as in trauma-induced coagulopathy. Thus, pharmacological inhibition of APC activity may improve blood clottability in certain clinical situations. In this study, we demonstrate that the DNA aptamer HS02-52G binds with fast onset (1.118 ± 0.013 × 105M-1s-1) to APC and possesses a long residence time of 13.5 min within the aptamer-APC complex. Functional analysis revealed HS02-52G as a highly potent and specific inhibitor of APC in plasma and whole blood with IC50values ≤30 nM, whose activity can be readily neutralized by the short complementary DNA molecule AD22. These features qualify the novel aptamer-antidote pair as a candidate treatment option for acute APC-related bleedings.
authors.affiliation: 1 Institute of Experimental Hematology and Transfusion Medicine, University of Bonn Medical Center , Bonn, Germany .
authors.forename: Nasim Shahidi
authors.initials: NS
authors.lastname: Hamedani
authors.affiliation: 1 Institute of Experimental Hematology and Transfusion Medicine, University of Bonn Medical Center , Bonn, Germany .
authors.forename: Heiko
authors.initials: H
authors.lastname: Rühl
authors.affiliation: 1 Institute of Experimental Hematology and Transfusion Medicine, University of Bonn Medical Center , Bonn, Germany .
authors.forename: Julia Janina
authors.initials: JJ
authors.lastname: Zimmermann
authors.affiliation: 2 Pall GmbH , Dreieich, Germany .
authors.forename: Tim
authors.initials: T
authors.lastname: Heiseler
authors.affiliation: 1 Institute of Experimental Hematology and Transfusion Medicine, University of Bonn Medical Center , Bonn, Germany .
authors.forename: Johannes
authors.initials: J
authors.lastname: Oldenburg
authors.affiliation: 3 Life and Medical Sciences Institute, University of Bonn , Bonn, Germany .
authors.forename: Günter
authors.initials: G
authors.lastname: Mayer
authors.affiliation: 1 Institute of Experimental Hematology and Transfusion Medicine, University of Bonn Medical Center , Bonn, Germany .
authors.forename: Bernd
authors.initials: B
authors.lastname: Pötzsch
authors.affiliation: 1 Institute of Experimental Hematology and Transfusion Medicine, University of Bonn Medical Center , Bonn, Germany .
authors.forename: Jens
authors.initials: J
authors.lastname: Müller
mesh_terms: D000925:Anticoagulants / Q000138:chemical synthesis / Q000737:chemistry*; D052157:Aptamers, Nucleotide / Q000138:chemical synthesis / Q000737:chemistry*; D020029:Base Pairing; D006801:Humans; D007700:Kinetics; D009690:Nucleic Acid Conformation; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000737:chemistry*; D010314:Partial Thromboplastin Time; D011485:Protein Binding; D011486:Protein C / Q000037:antagonists & inhibitors* / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D013816:Thermodynamics; D013917:Thrombin / Q000819:agonists / Q000037:antagonists & inhibitors / Q000737:chemistry*; D014914:Whole Blood Coagulation Time
chemical_list: D000925:Anticoagulants; D052157:Aptamers, Nucleotide; D016376:Oligonucleotides, Antisense; D011486:Protein C; D011994:Recombinant Proteins; D013917:Thrombin; C421124:drotrecogin alfa activated
keywords: activated protein C (APC); antidote; aptamer; hemophilia; inhibitor; trauma-induced coagulopathy (TIC)