Synthesis and testing of DNA bending agents

DNA弯曲剂的合成与测试

基本信息

批准号：
6702937
负责人：
Steven M Firestine
金额：
$ 21.64万
依托单位：
DUQUESNE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-06-01 至 2007-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6702937
关键词：
DNA binding protein DNA repair DNA topoisomerases amides gel mobility shift assay gene expression nuclear magnetic resonance spectroscopy nucleic acid chemical synthesis organic chemicals polymerase chain reaction polymers synthetic nucleic acid

项目摘要

DESCRIPTION (provided by applicant): A number of well-known and well-investigated DNA binding drugs have been shown to induce bends in DNA. Studies on the biological affects of these agents have indicated that DNA bending by these agents plays an important role in their mechanism of action. Examination of the literature has suggested that there are at least four mechanisms by which DNA bending produces a biological affect. The four mechanisms are: 1. induction of DNA repair resulting in induced strand breakage; 2. facilitating DNA cleavage by binding to topoisomerases; 3. inhibit the binding of transcriptional proteins; 4. facilitating the binding of transcriptional proteins. In addition, the integral role that DNA bending plays in the regulation of gene expression indicates that DNA bending agents may be able to regulate the expression of disease-associated genes. Unfortunately, currently available agents are not sequence specific and therefore cannot be targeted to a specific region of the human genome. The creation of sequence specific DNA bending agents is hampered by the fact that the design principles for preparing DNA bending agents are non-existent in the chemical literature. To date, there has been only one example of a rationally designed DNA bending agent and this agent consisted of a triple helix making "drug" delivery difficult. The purpose of this grant is to explore the design principles that would lead to the synthesis of a DNA bending agents. It is envisioned that this goal will provide utility in the creation of new medicinal agents that function by one of the five mechanisms outlined above. We will utilize the agents prepared in this proposal to investigate their ability to control transcription of simple model systems, as well as more complex systems related to angiogenesis. To accomplish this goal, we will leverage a combination of techniques from organic synthesis to molecular biology to focus on three specific aims. Specific Aim I: The design, synthesis and evaluation of polyamide-based DNA bending agents that bind in a 1:1 complex with DNA; Specific Aim II: Incorporation of DNA bending agents into polyamides that can target a specific gene in the human genome; Specific Aim III: The design and synthesis of DNA bending agents that "pull" the DNA together.

描述（由申请人提供）：许多众所周知且经过充分研究的 DNA 结合药物已被证明可以诱导 DNA 弯曲。对这些药物的生物学影响的研究表明，这些药物引起的 DNA 弯曲在其作用机制中起着重要作用。对文献的审查表明，DNA 弯曲至少有四种机制可以产生生物学影响。这四种机制是： 1. 诱导 DNA 修复，导致诱导链断裂； 2.通过与拓扑异构酶结合促进DNA切割； 3.抑制转录蛋白的结合； 4.促进转录蛋白的结合。此外，DNA弯曲在基因表达调节中发挥的不可或缺的作用表明DNA弯曲剂可能能够调节疾病相关基因的表达。不幸的是，目前可用的试剂不是序列特异性的，因此不能靶向人类基因组的特定区域。由于化学文献中不存在制备 DNA 弯曲剂的设计原理，因此阻碍了序列特异性 DNA 弯曲剂的创建。迄今为止，只有一个合理设计的 DNA 弯曲剂的例子，这种弯曲剂由三螺旋组成，使得“药物”输送变得困难。这笔资助的目的是探索合成 DNA 弯曲剂的设计原理。预计这一目标将为创造通过上述五种机制之一发挥作用的新药剂提供实用性。我们将利用本提案中制备的试剂来研究它们控制简单模型系统以及与血管生成相关的更复杂系统的转录的能力。为了实现这一目标，我们将利用从有机合成到分子生物学的技术组合来重点实现三个具体目标。具体目标 I：设计、合成和评估与 DNA 结合成 1:1 复合物的聚酰胺基 DNA 弯曲剂；具体目标二：将 DNA 弯曲剂掺入聚酰胺中，可以针对人类基因组中的特定基因；具体目标 III：设计和合成将 DNA“拉”在一起的 DNA 弯曲剂。