Synthetic and Analytical Methods Targeting Telomerase

靶向端粒酶的合成和分析方法

基本信息

批准号：
7211402
负责人：
SIMON H FRIEDMAN
金额：
$ 17.9万
依托单位：
UNIVERSITY OF MISSOURI KANSAS CITY
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-04-01 至 2009-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7211402
关键词：
Affinity Binding Biological Assay Catalysis Cell Line Chemistry Class Complex DNA DNA-Directed RNA Polymerase Depth Development Dissociation Enzyme Inhibitor Drugs Enzyme Inhibitors Enzymes Exhibits Generations Generic Drugs Genes Genomics Human Genome Intercalating Agents Kinetics Lead Libraries Ligands Malignant Neoplasms Methods Monitor Nucleic Acid Binding Nucleic Acids Nucleotides Numbers Pattern Phase Polymerase Prevention Proteins Purpose RNA Binding RNA-Directed DNA Polymerase Ribonuclease H Side Solid Somatic Cell Specificity Structure Surface Techniques Telomerase Telomerase Inhibitor Telomerase inhibition Validation Variant Work analytical method anti-cancer therapeutic base cancer cell combinatorial combinatorial chemistry design high throughput screening inhibitor/antagonist member novel strategies nucleic acid binding protein rapid technique technique development tool

项目摘要

DESCRIPTION (provided by applicant): The major long term objective of this application is to establish a new strategy to inhibit the enzyme telomerase. Telomerase is a promising universal anti-cancer target. Its activity is found in a majority of cancer cells and immortal cell lines, while being absent from a majority of healthy somatic cells. In addition to this correlation there are strong mechanistic reasons why cancer cells require telomerase. The central hypothesis of this project is that molecules which can bind the RNA/DNA duplex formed during telomerase's catalytic cycle will act as inhibitors of the enzyme, and will therefore be potential anti-cancer therapeutics. The postulated mechanism by which these molecules will inhibit the enzyme is either through the prevention of strand dissociation, a key step in telomerase's catalytic cycle, or by the distortion of the duplex substrate, leading to poor catalysis. Our preliminary studies have demonstrated that known RNA/DNA duplex binding molecules are able to inhibit telomerase, and do so in a manner consistent with this inhibition being due to interaction with the RNA/DNA duplex. The specific aims of this work are: 1) Assess known duplex binding molecules such as intercalators for activity as telomerase inhibitors. This assessment will include in-depth kinetic analysis of the mode of inhibition to definitively characterize the mechanism of action of the compounds 2) Use the most successful compounds as the basis of diverse libraries of compounds synthesized using combinatorial chemistry. The purpose of this is to introduce new moieties into the molecule, which will then introduce specific interactions with the unique surrounding telomerase surfaces. 3) Develop high throughput assays, including affinity methods and enzyme assays, which we will use to identify molecules from the combinatorial libraries with the highest affinity for telomerase. These techniques will allow the rapid assessment of large numbers of compounds, allowing a small group of high affinity compounds to be isolated from a large mixture. Combined, these three aims will allow the validation of a new approach for telomerase inhibition and the development of high specificity, high affinity lead inhibitors.

描述（由申请人提供）：本申请的主要长期目标是建立一种抑制酶端粒酶的新策略。端粒酶是一个有希望的通用抗癌靶标。它的活性是在大多数癌细胞和不朽的细胞系中发现的，而大多数健康的体细胞都没有。除了这种相关性外，癌细胞需要端粒酶的强烈机理原因。该项目的中心假设是可以结合端粒酶催化周期中形成的RNA/DNA双链体的分子将充当酶的抑制剂，因此将是潜在的抗癌疗法。这些分子将抑制酶的假定机制是通过预防链解离，这是端粒酶催化循环的关键步骤，或者是通过双链底物的变形，导致催化不良。我们的初步研究表明，已知的RNA/DNA双链结合分子能够抑制端粒酶，并以与RNA/DNA双链体相互作用而与这种抑制相一致的方式进行抑制。这项工作的具体目的是：1）评估已知的双链结合分子，例如用于端粒酶抑制剂的活性的介导子。该评估将包括对抑制模式的深入动力学分析，以确定表征化合物的作用机理2）使用最成功的化合物作为使用组合化学合成化合物的各种化合物库的基础。这样做的目的是将新部分引入分子，然后将其引入与独特的周围端粒酶表面的特定相互作用。 3）开发高吞吐量测定，包括亲和力方法和酶测定，我们将用它们从组合库中鉴定出对端粒酶最高亲和力的分子。这些技术将允许对大量化合物进行快速评估，从而可以从大型混合物中分离出一小部分高亲和力化合物。这三个目标结合在一起，将允许验证一种新的端粒酶抑制方法和高特异性，高亲和力铅抑制剂的发展。