Early Stage Discovery of Natural Products Targeting Anaerobic Protozoal Pathogen

针对厌氧原虫病原体的天然产物的早期发现

• 批准号: 9088344
• 负责人: Robert Henry Cichewicz
• 金额: $ 19万
• 依托单位: UNIVERSITY OF OKLAHOMA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-12 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9088344
• 关键词: Address; Adverse effects; Affect; Alcohols; American; Back; Biological; Biological Assay; Case Study; Cells; Chemicals; Clinical; Cyclosporins; Data; Detection; Development; Disease; Engineering; Entamoeba histolytica; Evaluation; Evolution; Exhibits; Face; Future; Giardia lamblia; Goals; HIV; Health; Human; In Vitro; Individual; Infection; Infectious Agent; Infertility; Inflammatory; Investigation; Libraries; Low Birth Weight Infant; Malignant neoplasm of cervix uteri; Mammalian Cell; Medical; Metronidazole; Mission; Morbidity - disease rate; Names; Natural Products; Neonatal Mortality; Neuraxis; Nitroimidazoles; Organism; Parasites; Pelvic Inflammatory Disease; Penicillins; Pharmaceutical Preparations; Phase; Predisposition; Prevalence; Protozoa; Research; Research Design; Resistance; Risk; Secure; Small Business Innovation Research Grant; Source; Staging; Stevens-Johnson Syndrome; Structure; System; Testing; Therapeutic Agents; Tinidazole; Toxic effect; Toxin; Trichomonas Infections; Trichomonas vaginalis; United States; United States National Institutes of Health; Woman; Work; attributable mortality; base; bioactive natural products; combat; drug discovery; fungus; inhibitor/antagonist; innovation; meetings; men; metabolic abnormality assessment; milligram; novel; novel therapeutics; pathogen; preclinical evaluation; programs; prostatitis; response; scaffold; screening; success; tool; transmission process

 DESCRIPTION (provided by applicant): Trichomonas vaginalis is the most prevalent sexually-transmitted infectious organism in the United States with roughly 7 million people affected. Individuals infected with T. vaginalis face an increased risk of cervical cancer, infertility, pelvc inflammatory disease, prostatitis, and susceptibility to HIV. The nitroimidazoles are the only class of drugs available to treat trichomoniasis, but they suffer from several serious side effects Additionally, the emergence and spread of resistance to nitroimidazoles is becoming a concern. Consequently, there is a critical need for the development of new therapeutic agents to combat T. vaginalis and the proposed studies, designed in response to this RFA, provide a mechanism for the discovery and development of bioactive molecules that meet this need. Vital to the success of a drug discovery program is access to an outstanding library of chemically diverse substances that are well suited to become early stage leads. Natural products meet this essential requirement with metabolites from fungi having not been previously screened against T. vaginalis. Thus, this work is innovative since we will couple a novel natural product source with a new high-content screening system to identify promising bioactive substances from a large fungal secondary metabolite library. During the R21 phase of this investigation, we will focus on the following objectives to secure new compounds for treating T. vaginalis infections: (i) implement a high-content screening system for the detection of natural products that inhibit T. vaginalis viability; (ii) test a one-of-a-kind library of pure natural products, pre-fractionated materials, and secondary-metabolite-enriched fungal extracts for inhibitors of T. vaginalis; (iii) employ a mammalian-cell-based counter-screen to remove non-selective toxins from screening; and (iv) use fungal taxonomic information and secondary metabolite dereplication tools to select extracts and fractions for further studies. The most promising agents emerging from the R21 studies will be subjected to detailed R33 studies via the following aims: (i) purify bioactive natural products from fungal fractions and extract mixtures, (ii) establish the structures of bioactive metabolites and secure milligram quantities for biological evaluation, (iii) develop a complimentary high-content screening assay to evaluate each hit molecule against additional anaerobic protozoan targets (Giardia intestinalis and Entamoeba histolytica), and (iv) conduct in vitro metabolism studies of the most promising hit molecules and test their metabolites for toxicity and bioactivity. The successful accomplishment of these aims is expected to yield a focused set of natural products that inhibit T. vaginalis viability with minimal toxicity to human cells. It is anticipated that 3-5 promising inhibitors will be obtained that will serve as bioactiv leads for immediate investigation during subsequent R01/SBIR studies. The focus on securing high-quality bioactive compounds during the R21/R33 study is anticipated to help generate first-in-class molecules with applications for treating T. vaginalis and other anaerobic protozoans.

 描述（由申请人提供）：阴道毛滴虫是美国最常见的性传播感染生物，大约有 700 万人受到感染，感染阴道毛滴虫的个人患宫颈癌、不孕不育、盆腔炎和前列腺炎的风险增加。和对艾滋病毒的易感性 硝基咪唑是唯一一类可用于治疗滴虫病的药物，但它们还存在一些严重的副作用，如出现和感染。硝基咪唑耐药性的传播正在成为一个令人担忧的问题，因此迫切需要开发新的治疗药物来对抗阴道毛滴虫，而针对该 RFA 设计的拟议研究为发现和开发阴道毛滴虫提供了一种机制。满足这一需求的生物活性分子对于药物发现计划的成功至关重要，是获得一个优秀的化学多样性物质库，这些物质非常适合成为早期先导化合物，而天然产物可以满足这一基本要求，而真菌的代谢物以前从未有过。筛选针对因此，这项工作具有创新性，因为我们将一种新型天然产物来源与新的高内涵筛选系统结合起来，从大型真菌次级代谢物库中鉴定出有前景的生物活性物质。重点关注以下目标，以确保获得用于治疗阴道毛滴虫感染的新化合物：（i）实施高内涵筛选系统来检测抑制阴道毛滴虫感染的天然产物。 阴道毛滴虫的活力；（ii）测试一种独一无二的纯天然产物、预分馏材料和富含次生代谢物的真菌提取物的库，以检测阴道毛滴虫的抑制剂；基于反筛选，从筛选中去除非选择性毒素；(iv) 使用真菌分类信息和次级代谢物去重复工具来选择提取物和级分以进行进一步研究。 R21研究将通过以下目标进行详细的R33研究：（i）从真菌组分中纯化生物活性天然产物并提取混合物，（ii）建立生物活性代谢物的结构并确保用于生物学评估的毫克量，（iii）开发一项免费的高内涵筛选试验，用于评估每种命中分子针对其他厌氧原生动物靶标（肠贾第鞭毛虫和溶组织内阿米巴）的效果，以及 (iv) 进行体外代谢研究的最有前途的命中分子并测试其代谢物的毒性和生物活性预计将产生一组集中的天然产物，其抑制阴道毛滴虫的活力，并且对人类细胞的毒性最小。将获得 5 种有前途的抑制剂，这些抑制剂将作为后续 R01/SBIR 研究期间立即研究的生物活性先导化合物。 R21/R33 研究期间的重点是确保高质量的生物活性化合物，预计将有助于生成具有应用价值的一流分子。用于治疗阴道毛滴虫和其他厌氧原生动物。