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序列水平上得到了解。控制特定基因的表达将提供最终的基因治疗。含有N-甲基吡咯和N-甲基咪唑氨基酸的小沟结合聚酰胺基于对核酸序列的识别表现出有前途的性能。我们最近证明了倍癌霉素A（Du）和Py-Im发夹聚酰胺的A段之间的杂交，选择性地烷基化450 bp DNA片段内的匹配序列之一。在该项目期间，开发了各种类型的序列特异性DNA结合剂，并用于调节基因表达。通过掺入L促进的高效DNA烷基化使我们能够开发一种新型的序列特异性DNA链间交联剂，其仅在ImImPy存在下在9-bp序列5 '-PyGGC（T/A）GCCPu处交联双链。 ...更多信息 3'.在开发有效的序列特异性烷基化发夹聚酰胺时，发现烷基化部分的选择及其在小沟中的位置非常重要。例如，通过四种不同的烷基化Py-Im发夹聚酰胺对DNA烷基化的比较研究清楚地表明，乙烯基接头的组成显著影响DNA烷基化反应性。此外，Im和乙烯基接头的新组合有效地在DNA烷基化试剂的序列特异性识别中靶向G-C碱基对。这些合成化合物烷基化预定的DNA序列以抑制转录，并且它们中的一些在筛选组测定中对某些癌细胞系具有显著不同的效力。本研究开发的DNA烷基化试剂为开发新型的基因转录调控生物试剂提供了一条很有前途的途径。我们目前正在研究靶向编码区的DNA烷基化发夹聚酰胺是否会抑制转录过程中聚合酶的延伸。提出了后基因组时代合理设计分子药物的方向之一。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