Sequence-Specific Alkylating Agent as A Novel Type of Antitumor Agent

随着人类基因组计划的完成，包括癌症、遗传病和病毒性疾病在内的许多疾病都可以从DNA序列水平上得到理解。对特定基因表达的控制将提供最终的基因治疗。含有N-甲基吡咯和N-甲基咪唑氨基酸的微小沟槽结合聚酰胺在识别核酸序列方面表现出良好的性能。我们最近证明了多卡霉素A(Du)的A节段与Py-Im发夹聚酰胺之间的杂交物，选择性地烷化了450-bp DNA片段中的匹配序列之一。在这个项目中，已经开发了各种类型的序列特异性DNA结合剂，并用于基因表达的调节。L的引入促进了高效的DNA烷基化，使我们开发出一种新型的序列特异性DNA链间交联剂，它只在9-BP序列5‘-PYGGC(T/A)GCCPU-…上存在ImPy的情况下才能使双链交联更多的3英尺。在开发有效的序列特异性烷基化发夹聚酰胺时，烷基化部分的选择及其在小沟槽中的位置被发现是非常重要的。例如，四种不同烷基化的Py-Im发夹聚酰胺对DNA烷基化反应的比较研究清楚地表明，乙烯基接头的组成对DNA烷基化反应活性有显著影响。此外，在DNA烷化试剂的序列特异性识别中，Im和乙烯基接头的新组合有效地作用于靶G-C碱基对。这些合成的化合物使预定的DNA序列烷基化以抑制转录，其中一些化合物在筛选小组试验中对某些癌细胞株具有显著不同的效力。本研究开发的有效DNA烷基化试剂为开发新型生物制剂控制基因转录提供了一条很有前途的途径。我们目前正在研究针对编码区的DNA烷基化发夹多酰胺是否抑制转录过程中聚合酶的延长。提出了后基因组时代分子医学合理设计的未来方向之一。较少

By the completion of the human genome project, many diseases including cancer, hereditary and viral diseases can be understood by the DNA sequence level. Control of the specific gene expression will provide ultimate gene therapy. Minor groove binding polyamides containing N-methylpyrrole and N-methylimidazole amino acids exhibit promising performance based on the recognition of nucleic acid sequences. We recently demonstrated that a hybrid between segment A of duocarmycin A (Du) and a Py-Im hairpin polyamide, selectively alkylates one of the matched sequences within a 450-bp DNA fragment. During this project various types of sequence-specific DNA binding agents have developed and used for the regulation of gene expression. The highly efficient DNA alkylation facilitated by the incorporation of L allowed us to develop a new type of sequence-specific DNA interstrand crosslinking agent that cross-links double strands only in the presence of ImImPy, at the 9-bp sequence 5'-PyGGC(T/A)GCCPu- … More 3'. In developing an efficient sequence-specific alkylating hairpin polyamide, the choice of alkylating moiety and its location in the minor groove was found to be very important. For instance, comparative study of DNA alkylation by four different alkylating Py-Im hairpin polyamides clearly demonstrated that the composition of the vinyl linker dramatically affects the DNA alkylating reactivity. Furthermore, a new combination of Im and the vinyl linker effectively acted to target G-C base pairs in the sequence-specific recognition by DNA alkylating reagents. These synthetic compounds alkylated predetermined DNA sequences to inhibit transcription, and some of them possessed significantly different potency for certain cancer cell lines in the screening panel assay. The effective DNA alkylating agent developed in the present investigation provides a promising approach for developing new types of biological agents to control gene transcription. We are currently investigating whether DNA alkylating hairpin polyamides targeting coding regions inhibit polymerase elongation during transcription. One of the future directions of rational design of molecular medicine in the post genome era has been proposed. Less