Compound library screening using Giardia labmblia high throughput viability assay

使用贾第鞭毛虫高通量活力测定筛选化合物库

• 批准号: 8069710
• 负责人: OSNAT HERZBERG
• 金额: $ 3.19万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-01 至 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8069710
• 关键词: 2-cyclopentyl-5-(5-isoquinolylsulfonyl)-6-nitro-1H-benzo(D)imidazole; Adverse effects; Biological Assay; Bioluminescence; Bioterrorism; Categories; Cell Survival; Cells; Centers for Disease Control and Prevention (U.S.); Chemicals; Chinese Hamster Ovary Cell; Collaborations; Development; Diarrhea; Disease Outbreaks; Drug resistance; Epithelial Cells; Exhibits; Genomics; Giardia; Giardia lamblia; Giardiasis; Growth; Human; Inhibitory Concentration 50; Intestines; Libraries; Luciferases; Mammalian Cell; Methods; Metronidazole; Organism; Outcome; Pharmaceutical Preparations; Recurrence; Screening procedure; Specificity; Structure; System; Tinidazole; United States; United States National Institutes of Health; base; cytotoxicity; high throughput screening; killings; miniaturize; pathogen; research study; text searching

DESCRIPTION (provided by applicant): Giardia lamblia is a human pathogen afflicting impoverished nations, and the most common cause of outbreaks of diarrhea in the United States. Giardia has been classified by the CDC as a category B bioterrorism organism. The favored anti giardiasis drugs in the US are metronidazole or tinidazole, compounds that have undesirable side effects. Moreover, increasing resistance to drug regimes and recurrence are a concern. It is thus clear that alternative drug treatments are needed. We have developed a bioluminescence Giarida viability assay based on the trophozoites ATP content that is suitable for high throughput screening and miniaturized the assay for compound screening in 1536-well plates. We propose to screen the NIH compound library and investigate the selectivity of hit compounds using initially CHO cells and later CACO-2 cells. The minimum lethal concentration of selective compounds exhibiting low IC50 will be determined by re- growth experiments. For a few top hits, identification of the Giardia target will be undertaken and if successful, advanced to structure-based compound optimization. PUBLIC HEALTH RELEVANCE: Giardia lamblia is a human pathogen afflicting billions of people annually and a category B bioterrorism organism. Giardiasis treatments have undesirable side effects, recurrence, and increasing drug resistance. To facilitate discovery of better drugs, we have developed a Giardia viability assay and miniaturized it for high throughput compound screening. This assay will be used to screen the NIH compound library and identify compounds that kill Giardia trophozoites.

描述（由申请人提供）：贾迪亚·兰布利亚（Giardia Lamblia）是一种遭受贫困国家的病原体，也是美国腹泻爆发的最常见原因。贾第鞭疾病人已被疾病预防控制中心（CDC）分类为B类生物恐怖生物体。美国有利的抗贾第鞭毛病毒是甲硝唑或替诺唑，具有不良副作用的化合物。此外，对药物体制和复发的耐药性增加引起了人们的关注。因此，很明显需要替代药物治疗。我们已经基于滋养体ATP含量开发了一种生物发光的贾拉达生存力测定法，该测定法适用于高吞吐量筛选，并将1536孔板中化合物筛选的测定法进行了微型化。我们建议筛选NIH化合物库，并使用最初的CHO细胞和后来的CACO-2细胞研究HIT化合物的选择性。表现出低IC50的选择性化合物的最低致命浓度将由重生实验确定。对于一些最高的热门单曲，将实现贾第鞭毛虫目标的识别，如果成功，则将其提升为基于结构的化合物优化。 公共卫生相关性：贾迪亚·兰布利亚（Giardia Lamblia）是一种人类的病原体，每年遭受数十亿人的痛苦和B类生物恐怖生物体。贾第鞭毛虫治疗具有不良的副作用，复发和耐药性的增加。为了促进发现更好的药物，我们开发了贾第鞭毛的生存力测定法，并将其用于高通量化合物筛选。该测定法将用于筛选NIH化合物库，并识别杀死贾第鞭毛虫的化合物。