SYNTHESIS OF SPECIFICALLY MODIFIED OLIGONUCLEOTIDES

本项目将探索多标记DNA的合成和应用 包括15N和13C标签的片段。借成立为法团 我们应该能够大幅增加多个标签的数量 可以从核磁共振实验中获得的信息。这个多- 标签方法将通过合并多个- 标记的单体，也通过加入不止一个标记的单体 单体。我们已经为准备工作制定的路线 单标记的15N脱氧核苷将作为起点 用于制备多标记化合物。新标签 目前的目标化合物如下： 1.[7-15N]-次黄嘌呤 2.[6，7-15N]-腺嘌呤 3.[6，7-15N]-脱氧腺苷 4.[3，6-15N]-腺嘌呤 5.[3，6-15N]-脱氧腺苷 6.[1，6，7-15N]-脱氧腺苷 7.[2-13C]-[2，3-15N]-脱氧鸟苷 8.[2-13C]-[1，2-15N]-脱氧鸟苷 9.[2-13C]-[1，2，7-15N]-脱氧鸟苷 10.[2-13C]-[2，3-15N]-O6-甲基脱氧鸟苷 11.[2-13C]-[1，2-15N]-O6-甲基脱氧鸟苷 12.[2-13C]-[1，2，7-15N]-O6-甲基脱氧鸟苷 这些化合物，连同[4-15N]-脱氧胞苷将被合并 变成核磁共振研究用的DNA片段。[1-15N]-、[2-15N]-和[6-15N]- 标签监测沃森-克里克脸的相互作用，而[7-15N]- 和[3-15N]-标签监测主槽和副槽中的相互作用。 [2-13C]原子将提供关于环电流效应的信息， 并将用于检测分子中的15N化学位移 15N标签上有不可交换的质子。 具体来说，我们将定义O6MeG.A和O6MeG.A中存在的碱基配对 O6MeG.G错配，碱基配对平行-DNA。我们会调查 药物.DNA和多肽.DNA相互作用，包括DNA和 配体。DNA氢键，以及发生在 药物对多肽的结合。此外，在每个系统中，我们都将 尝试将氢键受体(O或N)的身份与 供体NH基团的化学位移变化。此外，有了这些 多标记化合物，我们也将能够监测变化 发生在病变部位、配体部位的水合作用 有约束力，或建筑群中的其他地方。

This project will explore the synthesis and use of multiply labeled DNA fragments that include both 15N and 13C labels. By incorporation of multiple labels we should be able to increase dramatically the amount of information that can be obtained from an NMR experiment. This multi- labeling approach will be accomplished both by incorporation of multiple- labeled monomers, and also by incorporation of more than one labeled monomer. The routes that we have already developed for preparation of the singly-labeled 15N deoxynucleosides will serve as a starting point for preparation of the multiply labeled compounds. The new labeled compounds targeted at this time are listed below: 1. [7-15N]-Hypoxanthine 2. [6,7-15N]-Adenine 3. [6,7-15N]-Deoxyadenosine 4. [3,6-15N]-Adenine 5. [3,6-15N]-Deoxyadenosine 6. [1,6,7-15N]-Deoxyadenosine 7. [2-13C]-[2,3-15N]-Deoxyguanosine 8. [2-13C]-[1,2-15N]-Deoxyguanosine 9. [2-13C]-[1,2,7-15N]-Deoxyguanosine 10. [2-13C]-[2,3-15N]-O6-Methyldeoxyguanosine 11. [2-13C]-[1,2-15N]-O6-Methyldeoxyguanosine 12. [2-13C]-[1,2,7-15N]-O6-Methyldeoxyguanosine These compounds, along with [4-15N]-deoxycytidine, will be incorporated into DNA fragments for NMR studies. The [1-15N]-, [2-15N]-, and [6-15N]- labels monitor interactions of the Watson-Crick face, while the [7-15N]- and [3-15N]-labels monitor interactions in the major and minor grooves. The [2-13C] atom will provide information about ring current effects, and will be used to detect the 15N chemical shift in molecules where there is non-exchangeable proton coupled to the 15N label. Specifically, we will define the base pairing present in O6MeG.A and O6MeG.G mismatches, and the base pairing in parallel-DNA. We will probe drug.DNA and peptide.DNA interactions, including both DNA.DNA and ligand.DNA H-bonding, as well as the exclusion of water that occurs upon drug on peptide binding. Further, in each of these systems, we will attempt to correlate the identity of the H-bond acceptor (O or N) with the chemical shift change of the donor NH group. In addition, with these multi-labeled compounds, we will also be able to monitor changes in hydration which occur at the site of the lesion, the site of ligand binding, or elsewhere in the complex.