硫代磷酸酯键的掺入最近已广泛用于治疗性寡核苷酸中。为了它们的分离和质量控制,需要新的高效、高灵敏度的分析方法。在这项工作中,一种新的亲和毛细管电泳方法已被开发并用于分离潜在的抗癌药物,2\',3'-环二腺苷二硫代磷酸酯(Rp,Rp)(ADU-S100),和三个最近新合成的二氟化衍生物的非对映异构体,3\',3'-环二(2'-氟,2'-脱氧腺苷硫代磷酸酯)。使用几种天然和衍生的环糊精(CD)作为BGE的手性添加剂,在pH范围5-9的各种背景电解质(BGE)中进行分离。用β-获得了相对良好的分离,γ-,和2-羟丙基-γ-CD在一些测试的BGE中。然而,使用2-羟丙基-β-CD手性选择剂在由40mMTris组成的BGE中以43.5mM平均浓度实现了最佳分离,40mMtricine,pH8.1。在这些条件下,所有先前的四个环状二核苷酸(CDN)在4分钟内进行基线分离.此外,测定上述BGE中所有四种CDN与2-羟丙基-β-CD的复合物的平均表观结合常数和平均实际离子迁移率。发现形成的复合物相对较弱,平均表观结合常数在12.2-94.1Lmol-1范围内,实际离子迁移率的范围为(-7.8至-12.7)×10-9m2V-1s-1。所开发的方法可用于分离,分析,以及上述和类似CDN的表征。
The incorporation of phosphorothioate linkages has recently been extensively employed in therapeutic oligonucleotides. For their separation and quality control, new high-efficient and high-sensitive analytical methods are needed. In this work, a new affinity capillary electrophoresis method has been developed and applied for the separation of a potential anticancer drug, 2\',3\'-cyclic diadenosine diphosphorothioate (Rp, Rp) (ADU-S100), and three recently newly synthesized diastereomers of its difluorinated derivative, 3\',3\'-cyclic di(2\'-fluoro, 2\'-deoxyadenosine phosphorothioate). The separation was performed in the various background electrolytes (BGEs) within a pH range 5-9 using several native and derivatized cyclodextrins (CDs) as chiral additives of the BGE. Relatively good separations were obtained with β-, γ-, and 2-hydroxypropyl-γ-CDs in some of the BGEs tested. However, the best separation was achieved using the 2-hydroxypropyl-β-CD chiral selector at 43.5 mM average concentration in the BGE composed of 40 mM Tris, 40 mM tricine, pH 8.1. Under these conditions, all the previous four cyclic dinucleotides (CDNs) were baseline separated within 4 min. Additionally, the average apparent binding constants and the average actual ionic mobilities of the complexes of all four CDNs with 2-hydroxypropyl-β-CD in the above BGE were determined. The formed complexes were found to be relatively weak, with the average apparent binding constants in the range of 12.2-94.1 L mol-1 and with the actual ionic mobilities spanning the interval (-7.8 to -12.7) × 10-9 m2 V-1 s-1. The developed method can be applied for the separation, analysis, and characterization of the above and similar CDNs.