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虫病，这是一种由原生动物寄生虫蓝氏贾第鞭毛虫引起的肠道感染。该药物的临床耐药性和反复感染的报告强调了替代疗法的必要性。由于肠道中的兰氏贾第虫滋养体增殖与贾第鞭毛虫病的症状密切相关，因此我们开发了一种小型化增殖测定法，适合高通量筛选抑制滋养体增殖的化合物。对约 1500 种生物活性化合物进行的初步筛选发现了 50 种小分子，可显着抑制或阻止生长。尚不清楚这些化合物中的许多是否能抑制兰氏革兰氏杆菌。我们建议使用这种简单的高通量测定来筛选大量化合物。该筛选将同时对两个 G.lambdalia 组合（基因型）进行，以鉴定抑制两个组合的化合物，以及差异干扰组合 A 或 B 增殖的化合物。在二次筛选中确认的抑制剂将根据其分子靶标进行分类，从而确定了 G. Lamplia 中新的药物途径。双重筛选将能够鉴定对感染人类的兰氏贾氏杆菌群落有不同影响的化合物。根据此类化合物的作用模式，我们将能够在组合之间表型差异的分子基础上提出假设。该提案是为了响应 R21 开发/探索性资助征集而提交的，该资助征集旨在支持处于早期阶段且基于新颖实验系统的项目。具体来说，我们将使用滋养体增殖测定法在 384 孔板中筛选具有 G.ambalia 组合 A 和组合 B 分离株的小分子文库。高通量筛选中鉴定出的抑制剂将通过额外的组合 A 和组合 B 分离株进行验证。显微镜和流式细胞术将用于研究选定的、已确认的抑制剂的靶标。公共卫生相关性：兰氏贾第鞭毛虫是肠道感染的常见原因。我们建议筛选大量小分子，以确定未来开发针对这种寄生虫的药物的新线索。