Preferential formation of (5S,6R)-thymine glycol for oligodeoxyribonucleotide synthesis and analysis of drug binding to thymine glycol-containing DNA
Preferential formation of (5S,6R)-thymine glycol for oligodeoxyribonucleotide synthesis and analysis of drug binding to thymine glycol-containing DNA
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DOI：
10.1093/nar/gkj443
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发表时间：
2006-01-01
影响因子：
14.9
通讯作者：
Iwai, S
中科院分区：
文献类型：
--
作者：
Shimizu, T;Manabe, K;Iwai, S
我们之前报道了含有胸腺嘧啶乙二醇的寡核苷酸的化学合成，这是DNA氧化损伤的主要形式。在制备酰胺磷构筑块的过程中，四氧化锇氧化保护胸腺嘧啶的主要产物是(5R,6S)-胸腺嘧啶乙二醇。为了获得另一种同分异构体(5S,6R)-胸腺嘧啶乙二醇的构建基块，以足够的量合成寡核苷酸，研究了Sharpless不对称二羟基化（AD）反应。虽然反应非常缓慢，但与（5R,6S）异构体相比，(5S,6R)-胸苷乙二醇优先得到。（5S,6R）-和(5R,6S)-胸苷二醇的比例为2:1，并形成反式异构体。以离子液体1-丁基-3-甲基咪唑六氟磷酸为助溶剂，在不改变偏好的情况下，反应速度加快，收率提高。制备了（5S,6R）胸腺嘧啶乙二醇的磷酰胺基块，合成了含有5S-胸腺嘧啶乙二醇的寡核苷酸。其中一个寡核苷酸通过圆二色（CD）光谱和表面等离子体共振（SPR）测量分析了二霉素A与含胸腺嘧啶二醇DNA的结合。双霉素A在AATT靶点内与含有胸腺嘧啶二聚体的双链结合，即使胸腺嘧啶二聚体与胞嘧啶相对放置，也能观察到其结合。
We previously reported the chemical synthesis of oligonucleotides containing thymine glycol, a major form of oxidative DNA damage. In the preparation of the phosphoramidite building block, the predominant product of the osmium tetroxide oxidation of protected thymidine was (5R,6S)-thymidine glycol. To obtain the building block of the other isomer, (5S,6R)-thymidine glycol, in an amount sufficient for oligonucleotide synthesis, the Sharpless asymmetric dihydroxylation (AD) reaction was examined. Although the reaction was very slow, (5S,6R)-thymidine glycol was obtained in preference to the (5R,6S) isomer. The ratio of (5S,6R)- and (5R,6S)-thymidine glycols was 2: 1, and a trans isomer was also formed. When an ionic liquid, 1-butyl-3-methylimidazolium hexafluorophosphate, was used as a co-solvent, the reaction became faster, and the yield was improved without changing the preference. The phosphoramidite building block of (5S,6R)thymidine glycol was prepared, and oligonucleotides containing 5S-thymine glycol were synthesized. One of the oligonucleotides was used to analyze the binding of distamycin A to thymine glycol-containing DNA by Circular dichroism (CD) spectroscopy and surface plasmon resonance (SPR) measurements. Distamycin A bound to a duplex containing either isomer of thymine glycol within the AATT target site, and its binding was observed even when the thymine glycol was placed opposite cytosine.