Screening Repurposing Libraries for the Identification of Drugs with Novel anti-Coccidioidal Activity

筛选再利用文库以鉴定具有新型抗球虫活性的药物

基本信息

批准号：
10541230
负责人：
Jieh-Juen Yu
金额：
$ 37.09万
依托单位：
UNIVERSITY OF TEXAS SAN ANTONIO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-01-01 至 2026-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10541230
关键词：
Acceleration Amphotericin B Animal Model Antifungal Agents Area Azole resistance Bioinformatics Biological Assay Cell Membrane Permeability Cell Wall Cells Central Nervous System Clinical Coccidioides Coccidioides immitis Coccidioides posadasii Coccidioidomycosis Complement Consumption Cytology Development Diameter Disease Drug Screening Drug Targeting Flow Cytometry Fluconazole Fluconazole resistance Gene Expression Genes Genetic Genomics Goals Grant Growth Health Hyphae Image In Vitro Incidence Infection Itraconazole Lead Libraries Life Cycle Stages Lower respiratory tract structure Lung Lung diseases Medical Medicine Meningitis Metabolic Methodology Methods Mexico Microscopy Modeling Molecular Molecular Mechanisms of Action Morphology Motivation Mus Mycoses New Agents Nuclear Pathway interactions Patients Persons Pharmaceutical Preparations Pharmacotherapy Phase Phenotype Positioning Attribute Property Public Health RNA analysis Reproduction spores Research Resolution Rodent Model Soil South America Symptoms System Testing The science of Mycology Therapeutic Therapeutic Index Time Toxic effect Transmission Electron Microscopy Treatment Efficacy United States Vaccines acute infection bench-to-bedside translation chemotherapy clinically relevant combat cost cost effective cytotoxicity desert fever drug candidate drug development experience fungus in vivo insight meter mouse model novel novel strategies pandemic response pharmacologic preclinical development resistant strain screening side effect statistics therapeutic development transcriptome transcriptome sequencing transcriptomics vaccine strategy

项目摘要

Summary (Project 1) Coccidioidomycosis is a fungal infection caused by Coccidioides posadasii and Coccidioides immitis. It is estimated that 150,000 new infections occur in the United States each year. The incidence of this infection continues to rise in endemic regions. There is an urgent need for the development of better therapeutic drugs against coccidioidomycosis. Currently, the management of coccidioidomycosis includes antifungal agents such as amphotericin B, fluconazole and itraconazole. However, their toxicity and side effects, and concerns over the rise of azole-resistant clinical isolates point to an urgent need to develop new agents to combat this disease. In this project, we propose to screen repurposing libraries in search for drugs with novel activity against parasitic spherules of C. posadasii, and to apply our newly developed fungal cytological profiling (FCP) methodology for the identification of potential anti-CM drug candidates. Coccidioides spp. have a unique dimorphic life cycle characterized by the conversion of arthroconidia to spherules to enter parasitic phase. Using spherules to screen drug library is more medically relevant compare to saprobic spores/hyphae. Alteration of critical factor and phenotype (e.g. isotropic growth, cell wall integrity, cell wall remodeling, multi- nuclear formation) associated with coccidioidal parasitic cycle development can be assessed by FCP for initial drug library screening and later for drug cellular action discovery. Lead compounds from the drug library screening will be characterized in vitro for their antifungal activity against relevant clinical Coccidioides isolates of both C. posadasii and C. immitis, including azole-resistant strains. Subsequent in vivo antifungal efficacy will be conducted using our newly developed Galleria mellonella model of coccidioidomycosis, which provides a robust and cost-effective pre-screening system before testing in a more laborious, time-consuming and expensive rodent models. The leading compounds will be further characterized to gain insights into their molecular mechanisms of action. Global transcriptomic analyses will identify putative drug-targeted genes/pathways, which will be further assessed by gene-specific deletion and complementation. This molecular assessment will be complemented by fungal phenotypic analyses including microscopy (confocal, TEM) and FCP to gain further insights into the mode of action of each drug candidate. The goal is to identify at least 4 leading repositionable compounds at the completion of this project for advancing to preclinical development for chemotherapy against coccidioidomycosis.

摘要（项目1）球孢子菌病是由波萨达球孢子菌和粗球孢子菌引起的真菌感染。这是据估计，美国每年新增感染病例达 15 万例。此次感染的发生率流行地区继续上升。迫切需要开发更好的治疗药物对抗球孢子菌病。目前，球孢子菌病的治疗包括抗真菌药物，例如如两性霉素 B、氟康唑和伊曲康唑。然而，它们的毒性和副作用以及人们对它们的担忧唑类抗药性临床分离株的增加表明迫切需要开发新药物来应对这种情况疾病。在这个项目中，我们建议筛选再利用文库以寻找具有新活性的药物对抗 C. posadasii 的寄生小球，并应用我们新开发的真菌细胞学分析 (FCP) 识别潜在抗 CM 候选药物的方法。球孢子菌属拥有独特的以节分孢子转变为小球并进入寄生期为特征的二态性生命周期。与腐生孢子/菌丝相比，使用小球筛选药物库在医学上更具相关性。关键因素和表型的改变（例如各向同性生长、细胞壁完整性、细胞壁重塑、多与球孢子寄生周期发育相关的核形成）可以通过 FCP 进行评估，以进行初步评估药物库筛选以及随后的药物细胞作用发现。药物库中的先导化合物将在体外筛选针对相关临床球孢子菌分离株的抗真菌活性 C. posadasii 和 C. immitis，包括唑类抗性菌株。随后的体内抗真菌功效将使用我们新开发的大蜡蚧球孢子菌病模型进行，该模型提供了在进行更加费力、耗时和昂贵的啮齿动物模型。主要化合物将被进一步表征，以深入了解其分子作用机制。全球转录组分析将确定假定的药物靶向基因/途径，将通过基因特异性删除和互补来进一步评估。这分子评估将得到真菌表型分析的补充，包括显微镜检查（共聚焦、 TEM）和 FCP，以进一步了解每种候选药物的作用模式。目标是确定该项目完成后至少有 4 种领先的可重新定位化合物可进入临床前阶段开发针对球孢子菌病的化疗。