Abstract
This study was designed to isolate and identify aptamer sequences, acting as thrombin antagonists, from defibrotide, a single-stranded DNA fraction. Two different aptamers were identified: 5′-GGTTGGATTGGTTGG-3′ and 5′-GGTTGGATCGGTTGG-3′. A third aptamer: 5′-GGATGGATCGGTTGG-3′ was found in the PCR product from the double-stranded defibrotide precursor. All aptamers were potent inhibitors of thrombin-induced platelet aggregation, thromboxane biosynthesis, Ca ++ and fibrin clot formation, effective concentrations being in the nanomolar range. There was no effect on U 46,619 (thromboxane mimetic) or collagen-induced responses. Similar effects were obtained with unfractionated defibrotide while the "nonsense" oligonucleotide 5′-GGTGGTGGTTGTGGT-3′ was inactive. The data suggest that antithrombin effects of defibrotide are caused by aptamers.
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