Topoisomerases: Target for Antitrypanosomal Therapy

拓扑异构酶：抗锥虫治疗的靶标

• 批准号: 6783633
• 负责人: Theresa A Shapiro
• 金额: $ 40.88万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-08-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6783633
• 关键词: DNA topoisomerases; RNA interference; SDS polyacrylamide gel electrophoresis; Trypanosoma brucei rhodesiense; X ray crystallography; chemical structure function; drug design /synthesis /production; drug screening /evaluation; enzyme activity; enzyme inhibitors; gene induction /repression; laboratory mouse; laboratory rat; molecular cloning; nucleic acid metabolism; parasitic disease chemotherapy; polymerase chain reaction; protozoal genetics; trypanosomiasis

African trypanosomiasis is lethal if not treated and its incidence is increasing. Existing therapies are antique by contemporary standards and no vaccine is available. The DNA topoisomerases are enzymes essential for nucleic acid biosynthesis and cell survival and are proven targets for clinically valuable anti-infective and antitumor drugs. Previous work from this lab has established that topoisomerase inhibitors cause dramatic lesions in nuclear and mitochondrial (kinetoplast) DNA which are directly proportional to cell killing. This is an ongoing project to examine the DNA topoisomerases as targets for antitrypanosomal drug development, and the specific aims are three. First is to study type I enzymes, including the intracellular localization of structurally unique topo IB. This aim also includes the newly identified topoisomerases IA, characterizing their gene sequence and investigating intrace!!ular enzyme function by RNAi silencing. Aim 2 focuses on a previously unrecognized, second type IIA topoisomerase that occurs in trypanosomes: its catalytic features and inhibitor susceptibilities, and the consequences of silencing by RNAi. The third aim is to bring these basic molecular studies closer to the clinic by exploring the structure-activity relationship and selective toxicity of known topoisomerase inhibitors against trypanosomes in vitro. Included are recently discovered inhibitors oftopo IB, as well as topo II inhibitors with representatives currently in clinical trials. Compounds that appear most promising will be evaluated in mice. These studies take a multi-faceted, rational, and tangible approach to the development of much-needed new anti-trypanosomal chemotherapy.

如果不治疗，非洲锥虫病是致命的，并且其发病率正在增加。现有疗法 按照当代标准是古董，没有疫苗可用。 DNA拓扑异构酶是对核酸生物合成和细胞存活必不可少的酶，并且已被证明是对 临床上有价值的抗感染和抗肿瘤药物。该实验室的先前工作已经建立 拓扑异构酶抑制剂在核和线粒体中引起剧烈病变（动型） 与细胞杀伤直接成正比的DNA。这是一个正在进行的项目，以检查 DNA拓扑异构酶作为抗胰腺药物发育的靶标，具体目的是三个。首先是研究I型酶，包括结构上的细胞内定位 独特的Topo IB。该目标还包括新确定的拓扑异构酶IA，表征 他们的基因序列和内在的！ular酶功能通过RNAi沉默。目标2 专注于以前未知的第二类IIA拓扑异构酶，发生在 锥虫：其催化特征和抑制剂敏感性以及 RNAi沉默。第三个目的是将这些基本的分子研究通过 探索已知拓扑异构酶的结构活性关系和选择性毒性 在体外针对锥虫的抑制剂。包括最近发现的抑制剂topo ib， 以及目前正在临床试验中的代表的Topo II抑制剂。出现的化合物 最有希望的人将在小鼠中评估。这些研究采用了多方面的，理性的和 急需的新抗肉质体化学疗法的开发方法。