Giardia drug targets: Structure, function and inhibitors

贾第鞭毛虫药物靶点：结构、功能和抑制剂

基本信息

批准号：
7347031
负责人：
OSNAT HERZBERG
金额：
$ 57.44万
依托单位：
UNIVERSITY OF MD BIOTECHNOLOGY INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-02-01 至 2010-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7347031
关键词：
Active Sites Adverse effects Affect Binding Biochemical Biological Biological Assay Biological Warfare Biotechnology Bioterrorism Categories Centers for Disease Control and Prevention (U.S.)Clinical Cloning Collaborations Complex Country Crystallization Databases Dependence Developed Countries Developing Countries Diarrhea Disease Outbreaks Drug Delivery Systems Enzyme Inhibitor Drugs Enzyme Inhibitors Enzymes Escherichia coli Evaluation Food Supply Genes Genetic Transcription Genome Genomics Giardia Giardia lamblia Giardiasis Goals Growth Human Immunocompetent Immunocompromised Host In Vitro Institutes Kinetics Laboratories Ligands Maryland Methods Molecular New Mexico Organism Parasites Patients Pharmaceutical Preparations Population Production Property Proteins Protozoa Range Rate Recombinant Proteins Recurrence Research Resistance Roentgen Rays Sequence Analysis Solubility Specificity Structure Techniques Testing United States United States National Institutes of Health Universities Validation Water Work X-Ray Crystallography analog base cofactor design foodborne genome sequencing improved inhibitor/antagonist insight multidisciplinary pathogen pre-clinical size waterborne

项目摘要

DESCRIPTION (provided by applicant): Giardia lamblia is a food and waterborne parasite prevalent in developing countries, and a major cause of outbreaks of diarrhea in the United Sates, that has been classified as a bioterrorism category B organism by the Center for Disease Control because compromised water and food supply could affect US populations as well as army units in foreign countries. Drugs commonly used against anaerobic protozoa are used to treat giardiasis. However, they produce undesirable side effects, clinical resistance may occur in both immunocompromised and immunocompetent patients, and the rate of recurrence is high. The goal of this project is to develop new drug targets for alternative treatments of giardiasis. The sequence of the G. lamblia genome is available, thus, a genomic approach combined with biological insight has been taken to identify enzymes essential for the survival of the organism that are absent or are sufficiently divergent from the human enzymes to exploit those differences in the design of inhibitors. The selected enzymes will be cloned and expressed in E. coli using high throughput techniques, and those that are expressed in soluble form will be validated in G. lamblia by transcription interference methods. Essential enzymes will be prepared for structural and functional studies. Crystal structures will be determined, and will serve to guide the design of inhibitors specific to the Giardia enzymes. Assays to determine the kinetic constants of the enzymes will be developed and the catalytic mechanism will be studied to facilitate the in vitro evaluation of the inhibitors. Crystal structures of enzyme/inhibitor complexes will reveal how the inhibitor may be further improved. The effect of the inhibitors on the growth of G. lamblia will be determined. The long range goals of the project is to establish a database of potential drug targets against giardiasis, to gain insight about their mechanism of action, and to prepare inhibitors that are ready for preclinical evaluation.

描述（由申请人提供）：贾尔迪亚·兰布利亚（Giardia Lamblia）是发展中国家普遍存在的食物和水性寄生虫，是联合国腹泻爆发的主要原因，由于疾病控制中心的疾病控制中心被归类为生物恐怖主义的B级生物体，因为水和食品供应受损可能会影响我们的人口众多国家和国外部队。通常使用针对厌氧原生动物的药物用于治疗贾第鞭毛病。但是，它们会产生不良的副作用，免疫功能低下和免疫能力患者可能会出现临床抵抗，并且复发率很高。该项目的目的是为贾第鞭毛疾病的替代治疗而开发新的药物靶标。可以使用G. lamblia基因组的序列，因此，已经采用了一种与生物学见解相结合的基因组方法，以识别对没有生物的生存的酶，这些酶与没有人类酶的生物生存至关重要，从而在抑制剂的设计中利用这些差异来利用这些差异。选定的酶将使用高吞吐量技术克隆并在大肠杆菌中表达，而以可溶性形式表达的酶将通过转录干扰方法在G. lamblia中进行验证。基本酶将准备进行结构和功能研究。将确定晶体结构，并将用于指导贾第鞭毛虫酶特异的抑制剂的设计。将开发确定酶动力学常数的测定法，并将研究催化机制以促进抑制剂的体外评估。酶/抑制剂复合物的晶体结构将揭示如何进一步改善抑制剂。将确定抑制剂对G. lamblia的生长的影响。该项目的远距离目标是建立一个针对贾第鞭毛疾病的潜在药物靶标数据库，以了解其作用机理，并准备准备临床前评估的抑制剂。