Inhibitors of poxvirus enzymes as novel drugs

抗痘病毒药物目标是一个紧迫的问题，因为人们担心未申报的天花库存 可能被用作生物恐怖武器。而根除天花则是预防的胜利 在免疫接种方面，天花的治疗从未超过支持性治疗。对其他人的治疗 人类的痘病毒感染(传染性软体病、猴痘和接种疫苗的并发症 痘苗病毒)也是非特异性的或根本不存在。我国人猴痘暴发疫情 2003年，美国中西部地区强调了再次出现人类痘病毒疾病的风险。 我们的目标是寻找治疗和化学预防天花的新药，通过阻断 病毒mRNAs的转录和封顶。我们将筛选LifePharms独特的专有库 从13,000个野生蘑菇中提取精华以发现针对牛痘病毒复制的新抑制剂 包装在传染性病毒粒子核心内的信使核糖核酸转录装置。体外转录 被渗透的病毒粒子的反应被普遍接受，以忠实地概括病毒早期的过程。 在宿主细胞中发生的信使核糖核酸的生物发生。通过体外筛选抑制信使核糖核酸合成和 通过渗透性病毒粒子的处理，我们希望找到可以阻断一个或多个 负责痘病毒早期mRNAs转录和封端的关键病毒酶。这一战略针对 与针对单个病毒蛋白的筛查相比，初级筛查具有关键优势，因为：(I)它选择 对于能够接近病毒核心内的靶的化合物；以及(Ii)它包含多个 单一分析平台内的潜在酶靶点。靶标包括：(I)依赖DNA的RNA 聚合酶；(Ii)ETF(早期转录因子)，一种依赖DNA的ATPase；(Iii)封闭酶RNA 三磷酸酶、RNA鸟苷酸转移酶和RNA鸟氨酸-N7甲基转移酶；(Iv)NPH1，转录 具有DNA依赖的ATPase活性的延伸/终止因子；(V)RNA解旋酶NPH2；和(Vi) DNA拓扑异构酶。这些痘病毒酶是突出的药物靶点和这些酶的特定抑制剂。 酶将为药物开发提供先导化合物，并为痘病毒的基础研究提供关键工具 复制。 虽然真菌天然产物在药理学和药物发现方面做出了重大贡献， 在所有真菌物种中，只有一小部分被筛选出生物活性化合物。生命制药公司的图书馆 包含的物种多样性几乎等同于之前检查过的所有真菌物种。药物的发现 这里的项目大纲融合了LifePharms，Inc.在真菌生态学和天然 产品提取物收购，斯图尔特·舒曼博士研究痘病毒酶学和分子生物学，以及 研究三角研究所天然产物药物9的发现和药物化学。

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.