A Rapid assay for RNA targeted drugs: Instrumentation Supplement

描述（由申请人提供）：治疗疾病的一些最重要的疗法靶向核酸。靶向核酸的药物包括抗癌药物、抗生素和 抗病毒药这些药物在全球的年销售额约为1390亿美元。靶向核酸的抗生素占所有抗生素年销售额420亿美元中的145亿美元，占前五类抗生素中的三种。抗生素耐药性危机导致了巨大的经济和生命成本。70%的感染性细菌对至少一种常用的抗生素治疗（NIH）具有耐药性。仅在美国，就有170万人在医院环境中感染，其中不到10万例导致死亡（NIH）。这种情况表明，开发新的抗生素疗法的需求增加。一种快速检测新型氨基糖苷类与A位点和其他核酸位点结合亲和力的方法将有助于发现有效的治疗方法。在第一阶段，我们已经开发了一种基于荧光的竞争分析，使用27个碱基的核糖体A位点的RNA模型和一种新的荧光报告分子，F-neo。随着更大的化合物文库的建立，该测定法容易适用于高通量形式。在第二阶段，我们的具体目标是1）开发一种基于荧光的自动化检测方法，用于筛选广泛的RNA靶点。这一目标包括将当前测定扩展到高通量自动化筛选，并使测定适应于包括其他核酸药物靶标。2)扩大氨基糖苷类结合分子库。第I阶段产生了一个小的内部化合物库，使用第I阶段开发的测定法对其进行筛选。在II期，通过使用各种连接体长度和类型将氨基糖苷类与不同类别的化合物缀合，将极大地扩展文库。3)筛选NUBAD化合物以测定和表征活性。该体内测定将有助于建立来自测定的“命中”与细胞中的活性之间的相关性。在这项工作的结论，我们将建立一个商业上可用的测定高通量筛选化合物的目标核酸。这项技术将填补高通量筛选行业的空白，并导致更有效地开发治疗与核酸疗法相关疾病的化合物。

DESCRIPTION (provided by applicant): Some of the most significant therapies that treat disease target nucleic acids. Drugs that target nucleic acid include cancer drugs, antibiotics, and antivirals. Combined these classes of drugs have annual sales worldwide of ~$139 billion. Antibiotics that target nucleic acids account for $14.5 billion of the $42 billion of sales annuall for all antibiotics, and account for three of the top five classes of antibiotics. The crisis of antibiotic resistance has resulted in a significant cost, both financially and in human life. Seveny percent of infectious bacteria are resistant to at least one commonly used antibiotic treatment (NIH). In the United States alone, 1.7 million people get an infection while in a hospital environment with just under 100,000 of these cases resulting in death (NIH). This situation demonstrates the increased need for the development of new antibiotic therapies. A rapid assay examining the binding affinity of novel aminoglycosides to the A-site and other nucleic acid sites will facilitate the discovery of effective therapies. In Phase I, we have developed a fluorescence based competition assay using a 27-base RNA model of the ribosomal A-site and a novel fluorescent reporter molecule, F-neo. This assay is readily adaptable to a high throughput format as larger compound libraries are established. In Phase II our specific aims are 1) develop an automated fluorescent based assay for screening broad based RNA targets. This aim includes expanding the current assay to a high throughput automated screen and adapting the assay to include other nucleic acid drug targets. 2) Expand the library of aminoglycoside conjugated molecules. Phase I resulting in a small in house compound library that was screened using the assay developed in Phase I. In Phase II the library will be greatly expanded by the conjugation of aminoglycosides with different classes of compounds using a variety of linker lengths and types. 3) Screening of NUBAD compounds to determine and characterize the activity. This in vivo assay will help establish a correlation between "hits" from the assay and activity in the cell. At the conclusion of this work, we will have established a commercially available assay for the high throughput screening of compounds that target nucleic acids. This technology will fill a niche in the high throughput screening industry, and result in more efficien development of compounds that treat disease related to nucleic acid therapies.