(< 0

(< 0.0001). When both aptamers were used collectively, each at a concentration Ixazomib citrate of 1000 nM for a total aptamer concentration of 2000 nM, the aPTT increased to 278 0.8 sec, which was greater than the aPTT of each aptamer individually or the DNA aptamer in conjunction with a mutant version of the TOG25 RNA aptamer (White et al. both exosites with the aptamers prospects to synergistic inhibition of thrombin-dependent platelet activation and procoagulant activity. This combination of exosite 1 and exosite 2 inhibitors Ixazomib citrate may provide a particularly effective antithrombotic approach. < 0.0001) (Fig. 3A). At a concentration of 2000 nM, the DNA aptamer improved the aPTT from a baseline of 32.3 0.1 sec to 143.5 4.5 sec, which decreased inside a dose-dependent manner to 44.8 0.3 sec at an aptamer concentration of 31.25 nM. At a concentration of 2000 nM, the RNA aptamer improved the aPTT to 100.7 0.8 sec, and like the DNA aptamer, decreased inside a dose-dependent manner to 41 sec. Open Ixazomib citrate in a separate window Number 3. Clotting activity with the DNA + RNA aptamers better than DNA or RNA only. (< 0.0001). When both aptamers were used collectively, each at a concentration of 1000 nM for a total aptamer concentration of 2000 nM, the aPTT increased to 278 0.8 sec, which was greater than the aPTT of each aptamer individually or the DNA aptamer in conjunction with a mutant version of the TOG25 RNA aptamer (White et al. 2001) that contains a single nucleotide substitution (Fig. 3A). This apparent synergistic effect of both aptamers was also seen at total concentration of 1000 nM. At 500 nM, however, there Rabbit polyclonal to INPP1 was no significant difference between the ideals of the sum of the aPTT of the DNA and RNA and both compounds tested collectively (166.4 2.6 sec versus 167.5 0.4 sec, = 0.72). At doses below 500 nM, the effect of both the DNA and RNA ligands in the aPTT remains greater than each aptamer only, but the synergistic effect is no longer observed (Fig. 3A). At concentrations below 32 nM the aptamers have minimal effect (Fig. 4A). Open in a separate window Number 4. Clotting activity with the DNA + RNA aptamers at concentrations <40 nM. (< 0.0001) (Fig. 3B). At a concentration of 2000 nM, the DNA aptamer improved the PT from a baseline of 13.2 0.1 sec to 82.0 0.8 sec. This decreased inside a dose-dependent manner to 14.1 0.2 sec, which was statistically insignificant from baseline (= 0.06). At a concentration of 2000 nM, the RNA aptamer nominally Ixazomib citrate improved the PT to 16.4 sec and at a concentration of 62.5 nM, had essentially returned to baseline (13.4 sec, = 0.18 compared with baseline). However, when both aptamers were tested collectively at 2000 nM total concentration, the PT was 177.6 0.4 sec. At concentrations below 31.25 nM, the PT for the DNA + RNA did not significantly change (Fig. 4B). The TCT is definitely a specific assay that actions the conversion of fibrinogen to fibrin in the presence of thrombin and is consequently sensitive to inhibitors that interfere with the catalytic activity of thrombin. A statistically significant difference was observed between the DNA (or DNA + mutRNA), RNA, and DNA + RNA aptamer within the TCT (< 0.0001); however, the picture was quite different from that observed in the additional two thrombin-sensitive clotting assays. The effect of the DNA aptamer on TCT was quite pronounced, and at concentrations of aptamer above 62.5 nM, the clotting time exceeded the top limit of the assay (>999 sec) (Fig. 3C). The RNA aptamer, on the other hand,.