A Rapid Assay for RNA Targeted Drugs

RNA 靶向药物的快速检测

基本信息

批准号：
8715189
负责人：
DEV PRIYA ARYA
金额：
$ 68.47万
依托单位：
NUBAD, LLC
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8715189
关键词：
Accounting Address Affinity Amikacin Amino Sugars Aminoglycosides Anti-Bacterial Agents Antibiotic Resistance Antibiotic Therapy Antibiotics Antineoplastic Agents Antiviral Agents Bacteria Bacterial Infections Binding Biological Assay Cells Cessation of life Clinical Collaborations Derivation procedure Dermal Development Disease Drug Targeting Environment Equilibrium Escherichia coli Fibroblasts Fluorescein Fluorescence Growth HIV Hospitals Housing Human Immune response Industry Infection Kanamycin Length Libraries Life Liquid substance Malignant Neoplasms Medical Modeling Neomycin Nucleic Acids Pathology Pharmaceutical Preparations Phase Preparation RNA Reader Reporter Research Ribosomes Robot Sales Site Skin South Carolina Structure System Technology Tobramycin Toxic effect United States United States National Institutes of Health Universities Work antimicrobial bacterial resistance base clinically relevant commercialization cost cytotoxicity deprotonation drug discovery effective therapy functional group high throughput screening in vivo keratinocyte nephrotoxicity next generation novel ototoxicity pathogen public health relevance resistant strain screening

项目摘要

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.

描述（由申请人提供）：治疗疾病靶核酸的一些最重要的疗法。以核酸为靶点的药物包括抗癌药物、抗生素和抗病毒药物。这些类别的药物在全球的年销售额合计约为 1,390 亿美元。在所有抗生素年销售额 420 亿美元中，针对核酸的抗生素占 145 亿美元，并且占前五类抗生素中的三类。抗生素耐药性危机造成了巨大的经济和生命损失。百分之七十的传染性细菌对至少一种常用抗生素治疗 (NIH) 具有耐药性。仅在美国，就有 170 万人在医院环境中受到感染，其中不到 10 万例导致死亡 (NIH)。这种情况表明对开发新抗生素疗法的需求不断增加。快速检测新型氨基糖苷类与 A 位点和其他核酸位点的结合亲和力将有助于发现有效的治疗方法。在第一阶段，我们开发了一种基于荧光的竞争测定法，使用核糖体 A 位点的 27 碱基 RNA 模型和新型荧光报告分子 F-neo。随着更大的化合物库的建立，该测定很容易适应高通量格式。在第二阶段，我们的具体目标是 1) 开发一种基于自动化荧光的检测方法，用于筛选广泛的 RNA 靶标。这一目标包括将当前的测定扩展到高通量自动筛选，并调整该测定以包括其他核酸药物靶标。 2) 扩展氨基糖苷结合分子库。第一阶段产生了一个小型的内部化合物库，该库是使用第一阶段开发的测定法进行筛选的。在第二阶段，该库将通过使用各种接头长度和类型将氨基糖苷类与不同类别的化合物缀合而大大扩展。 3) 筛选 NUBAD 化合物以确定和表征活性。这种体内测定将有助于建立测定的“命中”与细胞活性之间的相关性。在这项工作结束时，我们将建立一种商业化的检测方法，用于高通量筛选靶向核酸的化合物。该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的疾病的化合物。