Kinetics of phosphorothioate oligonucleotide metabolism in biological fluids
Kinetics of phosphorothioate oligonucleotide metabolism in biological fluids
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DOI：
10.1093/nar/25.18.3615
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发表时间：
1997-09-15
影响因子：
14.9
通讯作者：
Cohen, AS
中科院分区：
文献类型：
--
作者：
Gilar, M;Belenky, A;Cohen, AS
使用毛细管电泳（CE）研究了GEM（R）91（一种与HIV-1 gag mRNA区域互补的25聚体硫代磷酸反义寡核苷酸）的体外稳定性和代谢。在37 ℃下，在人血浆中120小时内跟踪母体化合物的体外降解，使用激光诱导荧光检测的CE方法能够检测5 '端完整代谢物，包括从生物流体中提取的母体化合物，因为认为主要代谢途径是通过3'-外切核酸酶活性，将该研究的结果与化合物在含有3 ′-核酸外切酶的溶液中的稳定性进行比较。使用连续一级反应的数值解来获得动力学参数，母体化合物的核酸外切酶消化，如使用自动CE-UV仪器测量的，在两种体外系统之间以及在体外和体内系统之间产生了惊人的相似性。
The in vitro stability and metabolism of GEM(R)91, a 25mer phosphorothioate antisense oligonucleotide complementary to the gag mRNA region of HIV-1, was investigated using capillary electrophoresis (CE), The in vitro degradation of the parent compound at 37 degrees C was followed over the course of 120 h in human plasma, A CE method using laser-induced fluorescence detection was able to detect 5'-end intact metabolites including the parent compound extracted from biological fluids, Because the primary metabolic pathway is believed to be via 3'-exonuclease activity, the results of this study were compared with the stability of the compound in a solution containing 3'-exonuclease, The numerical solution of sequential first-order reactions was used to obtain kinetic parameters, Exonuclease digestion of the parent compound, as measured using an automated CE-UV instrument, yielded striking similarities between the two in vitro systems as well as between in vitro and in vivo systems.