High-throughput screening for new inhibitors of Giardia lamblia

兰氏贾第鞭毛虫新型抑制剂的高通量筛选

基本信息

批准号：
7936904
负责人：
GIOVANNI WIDMER
金额：
$ 20.57万
依托单位：
TUFTS UNIVERSITY BOSTON
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-21 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7936904
关键词：
Affect Biological Assay Bioterrorism Categories Cellular biology Charge Chemistry Clinical Collection Communities Country Cyst Data Databases Deposition Depressed mood Development Disease Drug Delivery Systems Drug resistance Emerging Communicable Diseases Equipment Exploratory/Developmental Grant Flow Cytometry Funding Future Genome Genotype Giardia Giardia lamblia Giardiasis Grant Growth High Prevalence Human Individual Infection Infectious Diseases Research Institutes Intestines Knowledge Laboratories Lead Letters Libraries Life Cycle Stages Liquid substance Logistics Metabolic Pathway Metronidazole Metronidazole resistance Microscope Microscopy Molecular Molecular Target National Institute of Allergy and Infectious Disease New England Parasites Pathogenesis Pathway interactions Pharmaceutical Preparations Pharmacotherapy Phase Preclinical Drug Evaluation Process Protozoa Reader Recurrence Relative (related person)Reporting Research Research Personnel Robotics Screening procedure Specificity Staging Symptoms System Taxon Therapeutic Time Viral Work base biodefense drug development high throughput screening inhibitor/antagonist interest medical schools miniaturize novel pathogen public health relevance response small molecule small molecule libraries

项目摘要

DESCRIPTION (provided by applicant): Metronidazole is commonly used for the treatment of giardiasis, an intestinal infection caused by the protozoan parasite Giardia lamblia. Reports of clinical resistance to this drug and recurrent infections underscore the need for alternative therapeutics. Since G. lamblia trophozoite proliferation in the intestine is intimately associated with the symptoms of giardiasis, we developed a miniaturized proliferation assay amenable to high-throughput screening for compounds which inhibit trophozoite proliferation. A pilot screen of ~1500 bioactive compounds uncovered 50 small molecules which significantly depressed or arrested growth. Many of these compounds were not known to inhibit G. lamblia. We propose to use this simple, high-throughput assay to screen a large collection of compounds. This screen will be conducted simultaneously with two G. lamblia assemblages (genotypes) to identify compounds which inhibit both assemblages, and compounds which differentially perturb proliferation of assemblage A or B. Inhibitors confirmed in a secondary screen will be categorized with respect to their molecular target, thus identifying new druggable pathways in G. lamblia. The dual screening will enable the identification of compounds which differentially affect G. lamblia assemblages infecting humans. Based on the mode of action of such compounds, we will be able to formulate hypotheses on the molecular basis of phenotypic differences between assemblage. This proposal is submitted in response to the R21 Developmental/Exploratory grant solicitation which is intended to support projects which are in their early phase and are based on novel experimental systems. Specifically, we will use a trophozoite proliferation assay to screen, in 384-well plates, small molecule libraries with an assemblage A and an assemblage B isolate of G. lamblia. Inhibitors identified in the high-throughput screen will be verified with additional assemblage A and assemblage B isolates. Microscopy and flow cytometry will be used to investigate the target of selected, confirmed inhibitors. PUBLIC HEALTH RELEVANCE: Giardia lamblia is a common cause of intestinal infections. We propose to screen large collections of small molecules to identify new leads for future development of drugs against this parasite.

描述（由申请人提供）：甲硝唑通常用于治疗贾第鞭毛病，这是由原生动物寄生虫Lamblia引起的肠道感染。对这种药物临床抵抗和复发感染的报道强调了对替代治疗的需求。由于肠道中的G. lamblia在肠道中的滋养体增殖与贾第鞭毛疾病的症状密切相关，因此我们开发了一种小型化的增殖测定法，可以对抑制滋养体增殖的化合物进行高通量筛查。〜1500个生物活性化合物的试验筛选发现了50个小分子，这些小分子显着降低或阻碍了生长。这些化合物中的许多尚未抑制G. lamblia。我们建议使用这种简单的高通量测定法来筛选大量化合物。该屏幕将与两种G. lamblia组合（基因型）同时进行，以鉴定抑制两种组合的化合物，并差异化合物A或B差异化合物B的抑制剂在二级筛选中确认的抑制剂将被分类为分子靶标，从而识别出可识别新药物的途径G. lamblia in lamblia。双重筛选将使能够鉴定化合物，这些化合物会差异地影响G. lamblia组合感染人类。基于此类化合物的作用方式，我们将能够根据分子基础表达组合之间的表型差异来提出假设。该提案是针对R21的发展/探索性授予招标提交的，该授予的旨在支持其早期阶段的项目，并基于新颖的实验系统。具体而言，我们将在384孔板中使用滋养体增殖测定法进行筛选，该板，带有A组合A的小分子库和G. Lamblia的组合B分离。在高通量屏幕中鉴定的抑制剂将通过其他组合A和组合B分离株进行验证。显微镜和流式细胞仪将用于研究选定的确认抑制剂的靶标。公共卫生相关性：贾迪亚·兰布利亚（Giardia Lamblia）是肠道感染的普遍原因。我们建议筛选大量的小分子，以确定针对这种寄生虫的未来开发药物的新潜在客户。