EXOCYCLIC ADDUCTS--SYNTHETIC AND ANALYTIC STUDIES

本项目开发的分析化学和有机合成 为该计划提供必要的基础。实验是 旨在鉴定由反应形成的化学修饰的核苷酸- 某些化疗药物和遗传毒性化合物的临床应用 原生DNA。交联剂的形成机制和序列特异性 将使用序列定义的双链寡核苷酸进行研究。 环外加合物、烷基化产物和交联型脱氧核苷酸 将使用先进的FAB质谱学检测技术进行鉴定- 计量、热喷雾LC/MS、核磁共振和高性能 和液相色谱学。 在其他实验中，有机合成导致单链 含有单一化学结构的碱基序列的寡聚脱氧核苷酸 提出了CALLI修饰部位的建议。该反应的已知外环产物 将合成含有BCNU、丙烯醛和氯乙烯的DNA， 转化成它们的亚磷酰胺并加入到合成的 使用固态技术的寡脱氧核苷酸。新的保护团 用于保护固态DNA中使用的核苷亚磷酰胺 合成将会被开发。这将允许包括碱- 合成低聚物中的敏感加合物。

The analytical chemistry and organic syntheses developed in this Project provide an essential underpinning for this Program. Experiments are designed to identify chemically-modified nucleotides formed by the reac- tions of certain chemotherapeutic agents and genotoxic compounds with native DNA. The mechanism and sequence specificity of cross-link formation will be investigated using sequence-defined duplex oligodeoxynucleotides. Exocyclic adducts, alkylation products, and cross-linked deoxynucleotides will be identified using advanced detection techniques of FAB mass spectro- metry, thermospray LC/MS, nuclear magnetic resonance, and high performance and liquid chromatography. In additional experiments, organic syntheses leading to single strand oligodeoxynucleotides of defined base sequence containing a single chemi- cally-modified site are proposed. Known exocyclic products of the reaction of DNA with BCNU, acrolein, and vinyl chloride, will be synthesized, converted to their phosphoramidites and incorporated into synthetic oligodeoxynucleotides using solid state techniques. New protecting groups for protection of the nucleoside phosphoramidites used in solid state DNA synthesis will be developed. This will permit the inclusion of the alkali- sensitive adducts into synthetic oligomers.