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Thrombin inhibitors based on single-stranded DNA aptamers.

Irina V Gribkova1, Vera A Spiridonova, Alexander S Gorbatenko

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Blood Coagulation & Fibrinolysis : an International Journal in Haemostasis and Thrombosis
|November 20, 2013
PubMed
Summary

Two DNA aptamers, 15TBA and 31TBA, effectively inhibit thrombin and act as anticoagulants. Aptamer 31TBA demonstrates superior efficacy, and both show stability in human blood, suggesting the liver clears them in vivo.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Pharmacology

Background:

  • Thrombin plays a crucial role in blood coagulation.
  • Aptamers are short DNA or RNA molecules with therapeutic potential.
  • Understanding aptamer stability and efficacy is vital for anticoagulant drug development.

Purpose of the Study:

  • To investigate the inhibitory effects of two DNA aptamers (15TBA and 31TBA) on fibrin polymerization.
  • To compare the anticoagulant properties of these aptamers using thrombodynamics.
  • To assess the in vitro stability of aptamers in human plasma and blood.

Main Methods:

  • In vitro measurement of thrombin inhibition via fibrin polymerization assays.
  • Assessment of anticoagulant activity using a thrombodynamics assay measuring spatial clot growth rate.
  • In vitro stability testing in human plasma and whole blood, monitoring activated partial thromboplastin time.

Main Results:

  • Both 15TBA and 31TBA dose-dependently inhibited fibrin polymerization and exhibited anticoagulant activity in human plasma.
  • Aptamer 31TBA was found to be 2-3 times more effective than 15TBA.
  • Both aptamers remained functionally stable in human plasma and whole blood for at least 4 hours in vitro.

Conclusions:

  • DNA aptamers 15TBA and 31TBA are potent inhibitors of thrombin and effective anticoagulants.
  • The stability of these aptamers in vitro suggests 3'-exonuclease is not responsible for their rapid clearance in vivo.
  • The liver is likely the primary organ responsible for the clearance of these oligonucleotide-based therapeutics from circulation.