Inhibitor Testing Core

抑制剂测试核心

基本信息

批准号：
8931202
负责人：
WILLIAM J STEINBACH
金额：
$ 22.64万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-06-25 至 2020-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8931202
关键词：
A549 Acute Address Algorithms Amphotericin B Anabolism Anidulafungin Animal Model Animals Antifungal Agents Aspergillosis Azoles Biological Biological Availability Biology Calcineurin inhibitor Candidiasis Caspofungin Cells Chemicals Chronic Clinical Clinical Trials Complex Coupled Cryptococcal Meningitis Cryptococcus neoformans infection Development Disease Ensure Erythrocytes Evaluation FDA approved Flucytosine Fungal Drug Resistance Grant Hemolysis Human Human Resources Immunosuppressive Agents In Vitro Invertebrates Knowledge Leadership Lipids Liposomes Lung Mammalian Cell Marketing Maximum Tolerated Dose Measures Methods Micafungin Modeling Modification Mucormycosis Mus Mycoses Nystatin Organism Pathogenesis Pathway interactions Polyenes Predisposition Program Research Project Grants Purine Antagonist Reproducibility Research Research Personnel Resistance development Resources Subcategory Testing The science of Mycology Tissues Toxic effect Translational Research Validation Variant Voriconazole base clinically relevant cytotoxicity fungus in vitro activity in vitro testing in vivo inhibitor/antagonist interest kidney cell meetings novel pathogen posaconazole pre-clinical programs research clinical testing respiratory

项目摘要

ABSTRACT: Inhibitor Testing Core Rigorous testing of novel inhibitors to define their efficacy against invasive fungal infections is the crux of this translational research. The Inhibitor Testing Core will conduct detailed in vitro and in vivo evaluations of the novel inhibitors arising from all three Projects for antifungal activity. This will unify the Projects by utilizing standard methods, materials, and facilities to depict the full scope and activity of each inhibitor. Fungal species present several inherent challenges to complete and accurate inhibitor assessment, including distinct disease subcategories and emergence of antifungal resistance in some species. Comprehensive evaluation therefore requires a prioritized approach by investigators knowledgeable in the nuances of fungal biology, clinical disease states, and currently available antifungals, coupled with a background in the discovery and development of antifungal compounds and strategies. The Inhibitor Testing Core provides robust expertise to support inhibitor validation, as its leadership has evaluated hundreds of compounds in vitro and in vivo and helped bring several to clinical trials or market. We will initially test novel inhibitors for in vitro antifungal activity by performing standardized, high-throughput evaluations of antifungal susceptibility for inhibitors from each of the three Projects. Subsequently, inhibitors of particular interest will be tested in a prioritized manner against a broader array of clinically-relevant fungi, as well as in combination with other antifungal agents to assess for synergistic activity. These synergy tests will be performed with FDA-approved antifungal classes (polyenes, azoles, echinocandins, flucytosine), with immunosuppressants (calcineurin inhibitors, Tor inhibitors, purine biosynthesis inhibitors), and with selected pathway inhibitors from other Projects in this proposal. We will then analyze mammalian toxicity profile of active novel inhibitors. To accomplish this, we will define the in vitro toxicity of active inhibitors as measured by mammalian cell cytotoxicity using human red blood cell hemolysis, followed by human respiratory cells (A549), and human kidney cells (HEK293). Secondary evaluation will define animal toxicity, including a murine maximum tolerated dose and evaluation of acute and chronic murine toxicities. Lastly, we will test inhibitors for efficacy in animal models of the major invasive fungal pathogens. Inhibitors showing activity in vitro, and acceptable toxicity, will be assessed in vivo using targeted animal models of the three major invasive fungal infections. The choice of animal models will be guided by in vitro antifungal activity against the various fungal species. Testing will be also prioritized to begin with invasive candidiasis, followed by invasive aspergillosis, and then proceed to cryptococcal meningitis/pulmonary cryptococcosis. Invertebrate models of mucormycosis will be utilized for Project 1 inhibitors when applicable. Evaluating novel inhibitors via broad testing and in multiple animal models would not be possible outside of the Inhibitor Testing Core or between unconnected grants.

摘要：抑制剂测试核心对新型抑制剂进行严格测试以确定其对抗侵袭性真菌感染的功效是这一问题的关键转化研究。抑制剂测试核心将对抑制剂进行详细的体外和体内评估来自所有三个抗真菌活性项目的新型抑制剂。这将通过利用统一项目标准方法、材料和设施来描述每种抑制剂的完整范围和活性。真菌种类对完整和准确的抑制剂评估提出了一些固有的挑战，包括不同的疾病子类别以及某些物种中抗真菌耐药性的出现。综合评价因此需要熟悉真菌生物学、临床的细微差别的研究人员采取优先方法疾病状态和目前可用的抗真菌药物，以及发现和治疗的背景抗真菌化合物和策略的开发。抑制剂测试核心提供了强大的专业知识支持抑制剂验证，因为其领导层已在体外和体内评估了数百种化合物，帮助将一些产品推向临床试验或市场。我们将首先测试新型抑制剂的体外抗真菌活性通过对每种抑制剂的抗真菌敏感性进行标准化、高通量评估三个项目。随后，将针对特别感兴趣的抑制剂以优先顺序进行测试更广泛的临床相关真菌，以及与其他抗真菌药物组合来评估协同活动。这些协同作用测试将使用 FDA 批准的抗真菌类药物（多烯、唑类、棘白菌素、氟胞嘧啶），与免疫抑制剂（钙调神经磷酸酶抑制剂、Tor 抑制剂、嘌呤生物合成抑制剂），以及来自本提案中其他项目的选定途径抑制剂。我们随后将分析活性新型抑制剂的哺乳动物毒性特征。为了实现这一目标，我们将定义体外使用人红细胞溶血通过哺乳动物细胞细胞毒性测量活性抑制剂的毒性，其次是人呼吸细胞（A549）和人肾细胞（HEK293）。二次评估将定义动物毒性，包括小鼠最大耐受剂量以及急性和慢性小鼠的评估毒性。最后，我们将测试抑制剂在主要侵入性真菌病原体动物模型中的功效。将使用目标动物在体内评估显示出体外活性和可接受的毒性的抑制剂三种主要侵袭性真菌感染的模型。动物模型的选择将以体外实验为指导针对各种真菌物种的抗真菌活性。测试也将优先从侵入性开始念珠菌病，然后是侵袭性曲霉病，然后进行隐球菌性脑膜炎/肺隐球菌病。如果适用，毛霉菌病的无脊椎动物模型将用于项目 1 抑制剂。在多种动物模型中通过广泛的测试来评估新型抑制剂是不可能的抑制剂测试核心或不相关的拨款之间。