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：抑制剂测试核心严格测试新型抑制剂以定义其针对侵入性真菌感染的功效是关键的翻译研究。抑制剂测试核心将在体外和体内进行详细的评估全部三个抗真菌活性项目引起的新型抑制剂。这将通过使用标准方法，材料和设施来描述每个抑制剂的全部范围和活动。真菌物种提出了完成和准确抑制剂评估的几个固有挑战，包括不同的疾病某些物种中抗真菌抗性的子类别和抗真菌抗性的出现。因此，全面评估需要研究人员在真菌生物学的细微差别方面的优先方法疾病状态以及目前可用的抗真菌剂，以及发现的背景开发抗真菌化合物和策略。抑制剂测试核心提供了强大的专业知识支持抑制剂验证，因为其领导层已经评估了数百种体外和体内化合物帮助几个临床试验或市场。我们最初将测试用于体外抗真菌活性的新型抑制剂通过对每种抑制剂的抗真菌敏感性进行标准化的高通量评估这三个项目。随后，将以优先的方式对特定感兴趣的抑制剂进行测试临床上与临床相关的真菌以及与其他抗真菌剂相结合的广泛阵列，以评估协同活动。这些协同测试将使用FDA批准的抗真菌类（Polyenes，叠唑，棘齿蛋白，氟胞嘧啶），含有免疫抑制剂（钙调神经酶抑制剂，TOR抑制剂，嘌呤生物合成抑制剂），并在该提案中其他项目中选择了途径抑制剂。然后我们会分析活性新型抑制剂的哺乳动物毒性概况。为此，我们将定义体外活性抑制剂的毒性通过使用人红细胞溶血测量的哺乳动物细胞毒性测量其次是人类呼吸细胞（A549）和人类肾细胞（HEK293）。次要评估将定义动物毒性，包括鼠的最大耐受剂量和急性和慢性鼠的评估毒性。最后，我们将测试主要侵入性真菌病原体动物模型中有效性的抑制剂。在体外表现出活性和可接受的毒性的抑制剂将在体内使用靶向动物进行评估三种主要侵入性真菌感染的模型。动物模型的选择将由体外指导针对各种真菌物种的抗真菌活性。测试还将优先考虑从侵入性开始念珠菌病，其次是浸润性曲霉病，然后继续进行隐孢菌脑膜炎/肺炎隐球菌病。适用的情况下，将用于项目1抑制剂的无脊椎动物模型。通过广泛的测试和多种动物模型评估新型抑制剂是不可能的抑制剂测试核心或未连接的补助金。