Development of a Gene Expression Control Medicine and Analysis of the Interaction with a DNA Duplex.

能够通过识别DNA序列特异性来控制基因表达的有机化合物有望成为可行的基因治疗药物。设计合成了含吡咯聚酰胺的核苷类化合物作为基因治疗药物的先导化合物，并用Tm值和圆二色谱分析了它们与DNA的相互作用。此外，还进行了可用于{(5R)-D-[5-^2H_1；5-^&lt；13&gt；C]ribofuranosyl}nucleoside构象结构分析的DNA的合成。1.用3-氨基丙基连接物(GAP)、氨丙基连接物(GBP)和含腺苷的吡咯聚酰胺(APY)合成了脱氧鸟苷基吡咯聚酰胺(GAP)。2.TM值和CD光谱对DNA双链的亲和力研究表明，与其他DNA络合物相比，含有5‘-AAATT-3’序列的DNA与GAP络合物具有较高的选择性。因此，GAP可能具有选择性地用于基因治疗的可用序列，为开发新的反义或抗基因药物提供了可能性。3.建立了{(5R)-D-[5-^2H_1；5-^&lt；13&gt；C]ribofuranosyl}thymine的高非对映选择性合成方法。用Ph_3P^&lt；13&gt；CH_3i-BuLi经Wittig反应合成了1，2：5，6-二-O-异丙基亚甲基-D-别乳糖的{(5R)-D-[5-^2H_1；5-^&lt；13&gt；C]核糖衍生物，并转化为D-[5-^&lt；13&gt；13&gt；α；13&gt；C-核糖衍生物，以AD反应、选择性酰化、NaIO_4氧化和从阿尔卑斯-硼烷-d到5-氧代-D-[5-^&lt；13&gt；C]核糖衍生物的立体选择性氢转移反应为主。以这种5-^&lt；13&gt；C/^2H_1-双标记核糖为原料，按既定方法合成了{(5R)-D-[5-^2H_1；5-^&lt；13&gt；C]ribofuranosyl}thymine。

Organic compounds capable of controlling gene expression by recognizing DNA sequence-specificity are expected to be viable gene therapy medicine. The nucleosides bearing pyrrolepolyamide which is minor groove binder were designed and synthesized as a lead compounds for gene therapy medicine and the analysis of DNA interaction with these compounds by Tm values and Circular Dichroism (CD) spectra was performed. In addition, the synthesis of {(5R)-D-[5-^2H_1;5-^<13>C]ribofuranosyl}nucleoside that is useful for structural analysis of conformation in DNA was carried out. 1.Deoxyguanosine bearing pyrrolepolyamide using 3-aminopropyl linker (GAP), using aminopropyonyl linker (GBP) and adenosine bearing pyrrolepolyamide (Apy) were synthesized. 2.The investigation of affinity for DNA duplexes by Tm values and CD spectra showed that DNA included the sequence of 5'-AAATT-3' with GAP complex has potential of highly selectivity as compare with complexes of other DNA. In this result, GAP might have usable sequence selectively in gene therapy, and provide possibility to be new series of antisense or antigene drugs. 3.Highly diastereoselective synthesis of {(5R)-D-[5-^2H_1;5-^<13>C]ribofuranosyl}thymine was established. A preparation of {(5R)-D-[5-^2H_1;5-^<13>C]ribose derivative of 1,2:5,6-di-O-isopropyliden-α-D-allofranose was successfully achieved by a ^<13>C Wittig reaction using Ph_3P^<13>CH_3I-BuLi to 5-oxoribose derivative and subsequent transformation into D-[5-^<13>C]ribose derivative with an AD reaction, selective acylation, oxidation with NaIO_4, and a stereoselective deuteride transfer reaction from Alpine-Borane-d to 5-oxo-D-[5-^<13>C]ribose derivative as the main reaction. {(5R)-D-[5-^2H_1;5-^<13>C]ribofuranosyl}thymine was synthesized in the established manner from this 5-^<13>C/^2H_1-double-labeled ribose.