Solid-phase synthesis of oligonucleotides containing a site-specific psoralen derivative
Solid-phase synthesis of oligonucleotides containing a site-specific psoralen derivative
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DOI：
10.1021/ja9703178
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发表时间：
1997-06-25
影响因子：
15
通讯作者：
Essigmann, JM
中科院分区：
文献类型：
--
作者：
Kobertz, WR;Essigmann, JM
香豆素是线性呋喃香豆素，用于治疗皮肤病1和皮肤T细胞淋巴瘤。2 DNA被认为是peptides治疗活性的细胞靶点。除了医学和DNA诊断应用外，含peptide的寡核苷酸也是确定核酸结构和功能的有用探针，3用于研究DNA修复机制，4和用于捕获RNA转录复合物。5在长波紫外光的作用下，卟啉通过[2+ 2]环加成反应，主要与双链DNA的5′-TA-3′位点上的胸苷形成光产物。该反应具有高度的区域和立体特异性，形成三种主要的具有顺式-顺式立体化学的光产物：支架间交联、吡喃酮侧单加合物和呋喃侧单加合物。6合成大量含紫杉醇的寡核苷酸的传统方法需要利用高强度激光，并且当所需的含紫杉醇的寡核苷酸仅含有一个5′-TA-3′位点时最好应用。[7]最好有一个全合成程序，允许在任何序列背景下制备peptide加合物，因为最近的研究表明，背景在DNA损伤剂的复制旁路、诱变和遗传毒性作用中起着重要作用。不能在任何所需的序列背景下设计寡核苷酸化的寡核苷酸限制了对这些重要治疗剂的生物学效应的研究。在这里，我们报道了2-羧基芘呋喃糖苷单加合物亚磷酰胺的合成及其与DNA的结合。首先，呋喃侧加合物含有完整的香豆素发色团，当与互补链杂交时，其可以在用长波UV光照射时转化为交联。第二，我们认为呋喃侧加合物可以在固相DNA合成中使用的碱性脱保护条件下存活，而已知吡喃酮侧单加合物的不饱和内酯在固相DNA合成中容易转移。
Psoralens are linear furocoumarins used for the treatment of skin diseases1 and cutaneous T-cell lymphoma. 2 DNA is believed to be the cellular target for the therapeutic activity of the psoralens. In addition to medical and DNA diagnostic applications, psoralen-containing oligonucleotides have been useful probes for determining nucleic acid structure and function, 3 for studying the mechanisms of DNA repair, 4 and for arresting RNA transcription complexes. 5 Psoralens react via a [2+ 2] cycloaddition, in concert with long-wave UV light, to form photoproducts primarily with thymidines in 5′-TA-3′ sites in double-stranded DNA. This reaction is highly regio-and stereospecific, forming three major photoproducts with cis-syn stereochemistry: an interstand cross-link, a pyrone-side monoadduct, and a furan-side monoadduct. 6 Traditional methods for the synthesis of large amounts of psoralen-containing oligonucleotides require the utilization of high intensity lasers and are best applied when the desired psoralen-containing oligonucleotide contains only one 5′-TA-3′ site. 7 It would be desirable to have total synthetic procedures allowing for the preparation of psoralen adducts in any sequence context because recent studies show that context plays an important role in the replicative bypass, mutagenic, and genotoxic effects of DNA-damaging agents. 8 The inability to design psoralenated oligonucleotides in any desired sequence context has restricted study of the biological effects of these important therapeutic agents. Here, we report the synthesis of a 2-carboxypsoralen furanside monoadduct phosphoramidite and its incorporation into DNA.Synthesis of the furan-side monoadduct was chosen for two reasons. First, the furan-side adduct contains an intact coumarin chromophore, which when hybridized to a complementary strand can be converted into the cross-link upon irradiation with longwave UV light. 9 Second, we believed that the furan-side adduct could survive the basic deprotection conditions used in solidphase DNA synthesis, whereas the unsaturated lactone of the pyrone-side monoadduct is known to transesterify readily in the