权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Stage-Specific Inhibitors of Orthopoxviruses

正痘病毒阶段特异性抑制剂

基本信息

批准号：
8257919
负责人：
John H Connor
金额：
$ 4.09万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
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：
2011-04-15 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8257919
关键词：
A549 Attenuated Biological Biological Assay Biological Models Bromides Cell Survival Cells Central Africa Chemicals Collaborations Combined Modality Therapy Communities DNA biosynthesis Data Development FDA approved Fluorescence Gene Expression Human Immunity Individual Infection Institutes Kinetics Laboratories Lead Libraries Licensing Life Cycle Stages Marketing Monitor Monkeypox Mosses Noise Orthopoxvirus Pharmaceutical Preparations Phase Population Poxviridae Poxviridae Infections Production Proteins Public Health Reporter Reporting Screening procedure Security Signal Transduction Smallpox Staging Structure-Activity Relationship System Testing Tetrazolium Therapeutic Agents Toxic effect Vaccination Vaccines Vaccinia Vaccinia virus Vaccinium Validation Venus Viral Viral Genes Virus Virus Diseases Virus Inhibitors Virus Replication Western Africa Work base cell killing cytotoxic cytotoxicity high throughput screening human disease improved inhibitor/antagonist interest killings novel pathogen programs prototype public health relevance recombinant virus small molecule tool viral DNA

项目摘要

DESCRIPTION (provided by applicant): We propose screening the MLPCN library to identify novel inhibitors of orthopoxviruses. Orthopoxviruses are a genus of viruses that include monkeypox, variola (the causative agent of smallpox) and vaccinia. Vaccinia is the prototypical orthopoxvirus which was used in the world-wide vaccination program that eradicated smallpox. Smallpox was once the most deadly human pathogen, and is estimated to have killed more than 300 million people. Following the eradication of smallpox, routine vaccination was discontinued in the 1970s and there has consequently been a precipitous decline in population immunity to smallpox and other orthopoxviruses. There are currently no FDA-licensed drugs to treat individuals infected with any poxvirus. Therapies for poxvirus infection are a significant priority given the threat of smallpox weaponization and the rise in reports of humans infected with monkey pox, which is endemic to Central and Western Africa and was exported to the US in 2003. The identification and development of new antipoxviral compounds will be of significant interest both to the public health community and to the Defense and Homeland Security agencies To identify new inhibitors of viral replication we will use the prototype orthopox virus, vaccine, as a screening virus. We have developed an HTS-ready screening approach based on vaccine-dependent expression of a rapidly maturing fluorescent protein. This assay has a signal-to-noise ratio of greater than 300:1 and allows kinetic as well as endpoint testing of replication. We have validated this assay through screening of two small- scale libraries. We will work with our collaborators at the Broad institute to couple this assay with a simple cell- viability assay that will be the basis of our large-scale screen for compounds that block virus replication without killing cells. To further classify relevant compounds we have developed and implemented a suite of assays using different reporter viruses for secondary screening that will allow validation and characterization of the initial screen "hits". These additional assays report on the individual stages of the vaccines life cycle and will allow us to "bin" the identified poxvirus inhibitors as inhibitors of early, intermediate or late stages of replication. We will work with our collaborators at the Broad Institute to develop and optimize the most interesting of our identified compounds (likely those that inhibit after viral DNA replication) towards high-efficacy low-toxicity antipoxviral compounds. Beyond the scope of this R03 application but of great interest to my lab and others in the community, probes will be tested for their abilities to inhibit the replication of other orthopox viruses (Monkey pox and Smallpox) through my laboratory's existing collaboration with USAMRIID. Through these efforts we will develop novel orthopox inhibitors capable of blocking Vaccine, Monkey pox, and Smallpox infection for application both as probes and as a therapeutic agent. PUBLIC HEALTH RELEVANCE: Project Narrative Poxviruses such as Smallpox and Monkeypox cause serious human disease. There are currently no FDA- approved drugs for treatment of these infections. We will use a highly attenuated cousin of these viruses called Vaccines virus to screen the MLPCN compound library for drugs that block poxvirus replication. Effective probes will be developed with the aim of providing new tools to further dissect the mechanism of viral infection and spread, and may also lead to one or more new drugs to protect against poxvirus infection.

描述（由申请人提供）：我们建议筛选MLPCN文库以鉴定新的正痘病毒抑制剂。正痘病毒是一种病毒属，包括猴痘、天花（天花的病原体）和牛痘。牛痘是一种典型的正痘病毒，曾用于世界范围内根除天花的疫苗接种计划。天花曾经是最致命的人类病原体，据估计已造成3亿多人死亡。在消灭天花之后，常规疫苗接种于1970年代停止，因此人口对天花和其他正痘病毒的免疫力急剧下降。目前还没有获得fda许可的药物用于治疗痘病毒感染者。鉴于天花武器化的威胁和人类感染猴痘报告的增加，痘病毒感染的治疗是一个重要的优先事项，猴痘是中非和西非的地方病，并于2003年出口到美国。识别和开发新的抗痘病毒化合物对公共卫生界，国防和国土安全机构来说都是非常重要的。为了识别新的病毒复制抑制剂，我们将使用痘病毒的原型，疫苗，作为筛选病毒。我们已经开发了一种基于疫苗依赖性表达的快速成熟荧光蛋白的hts筛选方法。该分析具有大于300:1的信噪比，并允许动力学和终点的复制测试。我们通过筛选两个小型文库验证了这一分析。我们将与Broad研究所的合作者合作，将这一试验与简单的细胞活力试验结合起来，这将是我们大规模筛选不杀死细胞而阻止病毒复制的化合物的基础。为了进一步对相关化合物进行分类，我们开发并实施了一套使用不同报告病毒进行二次筛选的检测方法，以验证和表征初始筛选的“命中”。这些附加检测报告了疫苗生命周期的各个阶段，并将使我们能够将已确定的痘病毒抑制剂“归类”为早期、中期或晚期复制阶段的抑制剂。我们将与Broad研究所的合作者合作，开发和优化我们发现的最有趣的化合物（可能是那些在病毒DNA复制后抑制的化合物），以实现高效、低毒的抗痘病毒化合物。超出了R03应用的范围，但我的实验室和社区其他人员对此非常感兴趣，探针将通过我的实验室与USAMRIID的现有合作，测试其抑制其他正痘病毒（猴痘和天花）复制的能力。通过这些努力，我们将开发出能够阻断疫苗、猴痘和天花感染的新型正痘抑制剂，既可以作为探针，也可以作为治疗剂。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The master regulator of the cellular stress response (HSF1) is critical for orthopoxvirus infection.

DOI：
10.1371/journal.ppat.1003904
发表时间：
2014-02
期刊：
PLoS pathogens
影响因子：
6.7
作者：
Filone CM;Caballero IS;Dower K;Mendillo ML;Cowley GS;Santagata S;Rozelle DK;Yen J;Rubins KH;Hacohen N;Root DE;Hensley LE;Connor J
通讯作者：
Connor J

Probing the virus host interaction in high containment: an approach using pooled short hairpin RNA.

探索高度遏制下的病毒宿主相互作用：一种使用混合短发夹 RNA 的方法。