Abstract
Purpose. To determine the degradation rates and pathways of GS-522, a potent oligodeoxynucleotide (GGTTGGTGTGGTTGG) inhibitor of thrombin, in serum and plasma.
Methods. A stability-indicating, anion-exchange HPLC method was developed and used to determine concentrations of GS-522 and metabolites.
Results. In monkey plasma at 2 µM or below, the degradation of GS-522 can be fit to a first-order exponential with a kp obs ~ 0.01 min−1. At 3 µM and above the degradation process deviates from a monoexponential decay profile. An initial fast degradation process is followed by a slower phase with an observed rate constant equal to that observed at 2 µM and below. In monkey serum, the KMand Vmaxare 8.4 µM and 0.87 µM min−1, respectively.
Conclusions. The kinetics are consistent with an equilibrium binding of GS-522 to prothrombin in plasma (Kd = 50 nM) which saturates at GS-522 concentrations >2 µM. Compared to a scrambled sequence (GGTGGTGGTTGTGGT), with no defined tertiary structure, GS-522 is 4-fold more stable in serum. The metabolic profile in plasma is consistent with a 3′-exonuclease catalyzed hydrolysis of GS-522.
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Shaw, JP., Fishback, J.A., Cundy, K.C. et al. A Novel Oligodeoxynucleotide Inhibitor of Thrombin. I. In VitroMetabolic Stability in Plasma and Serum. Pharm Res 12, 1937–1942 (1995). https://doi.org/10.1023/A:1016243923195
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DOI: https://doi.org/10.1023/A:1016243923195