In Vitro Evaluation of Aptamer-Based Reversible Inhibition of Anticoagulant Activated Protein C as a Novel Supportive Hemostatic Approach

Nucleic Acid Ther. 2016 Dec;26(6):355-362. doi: 10.1089/nat.2016.0645. Epub 2016 Oct 13.

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 × 105 M-1 s-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 IC50 values ≤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.

Keywords: activated protein C (APC); antidote; aptamer; hemophilia; inhibitor; trauma-induced coagulopathy (TIC).

MeSH terms

  • Anticoagulants / chemical synthesis
  • Anticoagulants / chemistry*
  • Aptamers, Nucleotide / chemical synthesis
  • Aptamers, Nucleotide / chemistry*
  • Base Pairing
  • Humans
  • Kinetics
  • Nucleic Acid Conformation
  • Oligonucleotides, Antisense / chemical synthesis
  • Oligonucleotides, Antisense / chemistry*
  • Partial Thromboplastin Time
  • Protein Binding
  • Protein C / antagonists & inhibitors*
  • Protein C / chemistry
  • Recombinant Proteins / chemistry
  • Thermodynamics
  • Thrombin / agonists
  • Thrombin / antagonists & inhibitors
  • Thrombin / chemistry*
  • Whole Blood Coagulation Time

Substances

  • Anticoagulants
  • Aptamers, Nucleotide
  • Oligonucleotides, Antisense
  • Protein C
  • Recombinant Proteins
  • Thrombin
  • drotrecogin alfa activated