Inhibitors of poxvirus enzymes as novel drugs

痘病毒酶抑制剂作为新药

基本信息

批准号：
7220568
负责人：
Esteban Edward Mena
金额：
$ 86.18万
依托单位：
LIFEPHARMS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-03-01 至 2010-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7220568
关键词：
Agaricales Antiviral Agents Ascomycota Basidiomycota Biogenesis Biological Assay Biological Factors Cells Chemoprophylaxis Cultured Cells DNA Topoisomerases DNA biosynthesis DNA-Directed RNA Polymerase DNA-dependent ATPase Disease Disease Outbreaks Drug Compounding Drug Delivery Systems Ecology Enzymatic Biochemistry Enzyme Inhibitor Drugs Enzyme Inhibitors Enzymes Equipment and supply inventories Genetic Transcription Goals Growth Guanine Human Immunization In Vitro Individual Infection Institutes Lead Libraries Mass Spectrum Analysis Messenger RNA Methodology Methyltransferase Midwestern United States Molecular Biology Molecular Target Molluscum Contagiosum Monkeypox Morphogenesis One-Step dentin bonding system Orthopoxvirus Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacology Poison Poisoning Poxviridae Poxviridae Infections Process Processed Genes Protein Biosynthesis RNA RNA Helicase RNA triphosphatase Reaction Research Risk Sampling Screening procedure Smallpox Source Structure Supportive care Transcription Elongation Vaccinia virus Viral Viral Proteins Virion Virus Replication X-Ray Crystallography analog base drug development drug discovery fungus inhibitor/antagonist mRNA guanylyltransferase novel particle prophylactic research study termination factor tool transcription factor

项目摘要

Antipoxvirus drug targets are a pressing issue, given the concern that undeclared stocks of smallpox might be used as a bioterror weapon. Whereas the eradication of smallpox was a triumph of prophylactic immunization, the treatment of smallpox never advanced beyond supportive therapy. Treatments for other poxvirus infections of humans (molluscum contagiosum, monkeypox, and complications of immunization with vaccinia virus) are also either nonspecific or nonexistent. The outbreak of human monkeypox infections in the US Midwest in 2003 highlighted the risks of re,emergence of human poxvirus disease. Our goal is to identify novel drugs for the treatment and chemoprophylaxis of smallpox by blocking the transcription and capping of viral mRNAs. We will screen LifePharms' unique and proprietary library of extracts from >13,000 wild mushrooms to discover new inhibitors of poxvirus replication targeted to the mRNA transcription apparatus packaged within the core of the infectious virion. The in vitro transcription reaction of permeabilized virions is generally accepted to faithfully recapitulate the process of viral early mRNA biogenesis as it occurs in the host cell. By screening in vitro for inhibition of mRNA synthesis and processing by permeabilized virions, we expect to identify candidate antivirals that block one or more of the key viral enzymes responsible for transcription and capping of poxvirus early mRNAs. This strategy for primary screening has key advantages over screens against individual viral proteins because: (i) it selects for compounds that are capable of accessing the target within the virion core; and (ii) it embraces multiple potential enzymatic targets within a single assay platform. The targets include: (i) DNA-dependent RNA polymerase; (ii) ETF (early transcription factor), a DNA-dependent ATPase; (iii) the capping enzymes RNA triphosphatase, RNA guanylyltransferase and RNA guanine-N7 methyltransferase; (iv) NPH1, a transcription elongation/termination factor with DNA-dependent ATPase activity; (v) NPH2, an RNA helicase; and (vi) DNA topoisomerase. These poxvirus enzymes are outstanding drug targets and specific inhibitors of these enzymes will provide lead compounds for drug development as well as key tools for basic studies of poxvirus replication. Although fungal natural products has made major contributions to pharmacology and drug discovery, only a fraction of all fungal species have been screened for bioactive compounds. LifePharms' library contains species diversity nearly equivalent to all fungal species examined previously. The drug discovery project outline here merges the complementary expertise of LifePharms, Inc. in fungal ecology and natural product extract acquisition, Dr. Stewart Shuman in poxvirus enzymology and molecular biology, and Research Triangle Institute in natural product dru 9 discovery and medicinal chemistry.

鉴于人们担心未申报的天花库存，抗痘病毒药物靶点是一个紧迫的问题可能被用作生物恐怖武器。鉴于消灭天花是预防性药物的胜利免疫接种、天花治疗从未超越支持疗法。其他治疗人类痘病毒感染（传染性软疣、猴痘以及免疫接种的并发症）痘苗病毒）也是非特异性的或不存在的。人类猴痘感染暴发 2003年美国中西部地区强调了人类痘病毒疾病再次出现的风险。我们的目标是通过阻断天花的传播来识别治疗和化学预防天花的新药病毒 mRNA 的转录和加帽。我们将筛选 LifePharms 独特的专有库从超过 13,000 种野生蘑菇中提取，发现针对痘病毒复制的新抑制剂 mRNA 转录装置包装在感染性病毒粒子的核心内。体外转录透化病毒粒子的反应被普遍认为忠实地再现了病毒早期的过程 mRNA 生物合成发生在宿主细胞中。通过体外筛选 mRNA 合成抑制通过透化病毒颗粒的处理，我们期望鉴定出能够阻断一种或多种病毒的候选抗病毒药物。负责痘病毒早期 mRNA 转录和加帽的关键病毒酶。这一策略对于与针对单个病毒蛋白的筛选相比，初级筛选具有关键优势，因为：（i）它选择对于能够接近病毒粒子核心内的靶标的化合物； (ii) 它包含多种单一检测平台内的潜在酶靶标。目标包括： (i) DNA 依赖性 RNA 聚合酶； (ii) ETF（早期转录因子），一种 DNA 依赖性 ATP 酶； (iii) 加帽酶 RNA 三磷酸酶、RNA鸟苷基转移酶和RNA鸟嘌呤-N7甲基转移酶； (iv) NPH1，转录具有 DNA 依赖性 ATP 酶活性的延伸/终止因子； (v) NPH2，RNA解旋酶；以及（六） DNA拓扑异构酶。这些痘病毒酶是杰出的药物靶点和这些酶的特异性抑制剂酶将为药物开发提供先导化合物以及痘病毒基础研究的关键工具复制。尽管真菌天然产物对药理学和药物发现做出了重大贡献，仅对所有真菌物种的一小部分进行了生物活性化合物筛选。 LifePharms 图书馆包含的物种多样性几乎与之前检查的所有真菌物种相同。药物发现此处的项目大纲融合了 LifePharms, Inc. 在真菌生态学和自然学方面的互补专业知识产品提取物采集，斯图尔特·舒曼博士在痘病毒酶学和分子生物学方面的研究，以及 Research Triangle Institute 从事天然产物 dru 9 发现和药物化学研究。