Improving Thermodynamic Stability and Anticoagulant Activity of a Thrombin Binding Aptamer by Incorporation of 8-trifluoromethyl-2′-deoxyguanosine
Improving Thermodynamic Stability and Anticoagulant Activity of a Thrombin Binding Aptamer by Incorporation of 8-trifluoromethyl-2′-deoxyguanosine
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DOI：
10.1021/acs.jmedchem.0c01711
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发表时间：
2021-01-14
影响因子：
7.3
通讯作者：
Xu, Yan
中科院分区：
文献类型：
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作者：
Bao, Hong-Liang;Ishizuka, Takumi;Xu, Yan
在这项研究中，我们将8-三氟甲基-2 '-脱氧鸟苷（（F）G）掺入凝血酶结合适体（TBA）中。通过圆二色谱、核磁共振、电泳和凝血酶原时间测定等方法研究了（F）G修饰TBA序列的结构、热力学稳定性、生物学稳定性和抗凝活性。我们发现，与天然序列相比，将（F）G替换为TBA序列导致解链温度显著提高，最高可达30摄氏度。三氟甲基使我们能够通过F-19 NMR光谱研究TBA G-四链体结构。PT试验表明，与天然TBA相比，修饰后的TBA具有更好的抗凝血活性。最后，我们证明了三氟甲基修饰的TBA序列可以在活体大鼠中作为抗凝剂发挥作用。我们的研究结果有力地表明，（F）G是一个强大的核苷衍生物，以增加TBA的热力学稳定性和抗凝血活性。
In this study, we incorporated 8-trifluoromethyl-2'-deoxyguanosine ((F)G) into a thrombin binding aptamer (TBA ). Circular dichroism, nuclear magnetic resonance (NMR), electrophoresis, and prothrombin time (PT) assay were performed to investigate the structure, thermodynamic stability, biological stability, and anticoagulant activity of the (F)G-modified TBA sequences. We found that the replacement of (F)G into TBA sequences led to a remarkable improvement in the melting temperature up to 30 degrees C compared with the native sequence. The trifluoromethyl group allowed us to investigate the TBA G-quadruplex structure by F-19 NMR spectroscopy. Furthermore, PT assays showed that the modified sequences can significantly improve the anticoagulant activity in comparison with the native TBA. Finally, we demonstrated that the trifluoromethyl-modified TBA sequence could function as an anticoagulant reagent in live rats. Our results strongly suggested that (F)G is a powerful nucleoside derivative to increase the thermodynamic stability and anticoagulant activity of TBA.