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

Discovery of Chemical Probes for RNA-Binding Protein Host Defense Factors

RNA 结合蛋白宿主防御因子化学探针的发现

基本信息

批准号：
9052780
负责人：
Harold C Smith
金额：
$ 60.56万
依托单位：
OYAGEN, INC.
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-15 至 2017-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9052780
关键词：
AIDS prevention APOCEC3G gene Acquired Immunodeficiency Syndrome Address Animal Model Antiviral Agents Attenuated Binding Biological Assay Biological Markers Cell model Cells Characteristics Chemicals Complex Custom DNA Deaminase Development Dimethyl Sulfoxide Diversity Library Dose Drug Compounding Drug Industry Drug Kinetics Drug Targeting Frequencies Future Glycerol Goals Grant HIV HIV Genome HIV Infections Harvest Health Host Defense Human Immune In Vitro Individual Infection Ions Lead Life Life Cycle Stages Modification Molecular Molecular Probes Monitor Multi-Drug Resistance Mus Mutate Mutation NIH Program Announcements Peripheral Blood Mononuclear Cell Pharmaceutical Chemistry Pharmaceutical Preparations Population Powder dose form Property Proteins RNA RNA Binding RNA-Binding Proteins Research Ribonucleoproteins Role Sedimentation process Series Shipping Ships Signal Transduction Technology Testing Therapeutic Index Tissues Toxic effect Triage United States National Institutes of Health Validation Viral Virion Virus Virus Diseases Western Blotting analog base cytotoxicity cytotoxicity test design efficacy testing high throughput screening improved in vivo innovation nanomolar next generation sequencing novel novel strategies particle prevent resistant strain response scaffold screening sexual HIV transmission small molecule transmission process treatment strategy uptake

项目摘要

DESCRIPTION (provided by applicant): This revised proposal is in response to the Program Announcement PA-12-060 entitled 'Solicitation of Validated Hits for the Discovery of in vivo Chemical Probes' (R01). The goal of this proposal is to synthesize and select novel chemical scaffolds as antagonists of APOBEC3G (A3G) binding to inhibitory RNA. These will serve as molecular probes to determine the role that RNA binding to A3G has in preventing host defense against an HIV infection. A3G is effective in host defense against HIV if it can evade destruction by the HIV protein known as Vif (made during late infection) and if it becomes packaged with viral particles and thereby enters cells with virions. The bulk of A3G preexisting in cells prior t an infection is largely inert as its activity is severely attenuated through its nonselective formation of ribonucleoprotein complexes with cellular RNAs. RNA binds to the N-terminus of A3G and allosterically inhibits the ssDNA binding and deaminase domain in the C-terminus. High throughput screening by OyaGen, Inc has identified three validated chemical scaffolds for their ability to: (1) antagonize RNA binding to A3G and (2) preemptively activate A3G in cells to the extent that incoming HIV replication is inhibited. In the proposed research OyaGen, Inc and Sanford/Burnham will collaborate to: (1) apply rational design to synthesize chemical modifications of three validated A3G- selective antiviral compounds, (2) select those with enhanced activity as antagonists of A3G:RNA binding and antiviral activity, (3) select the ultimate probe(s) based on enhanced activation of A3G DNA mutagenic activity and selectivity for cellular A3G-RNA complexes, and. (4) triage compounds to select those with drug-like characteristics based on in vitro ADME/T and in vivo PK in mice. These probes will usher in a significant paradigm change as they will enable studies and monitoring of the mechanism of action of APOBEC3G as it naturally is expressed in human cells during a live virus infection and, in the future, in animal models.

描述（由申请人提供）：本修订提案是对标题为“体内化学探针发现的验证命中征集”（R01）的计划公告 PA-12-060 的回应。该提案的目标是合成和选择新型化学支架作为 APOBEC3G (A3G) 与抑制性 RNA 结合的拮抗剂。这些将作为分子探针来确定 RNA 与 A3G 结合在阻止宿主防御 HIV 感染方面的作用。如果 A3G 能够逃避 HIV 蛋白 Vif（在感染后期产生）的破坏，并且它与病毒颗粒一起包装，从而进入带有病毒颗粒的细胞，那么 A3G 就可以有效地防御 HIV。感染前细胞中预先存在的 A3G 大部分基本上是惰性的，因为其活性通过与细胞 RNA 非选择性形成核糖核蛋白复合物而严重减弱。 RNA 与 A3G 的 N 末端结合，并以变构方式抑制 C 末端的 ssDNA 结合和脱氨酶结构域。 OyaGen, Inc 的高通量筛选已鉴定出三种经过验证的化学支架，它们具有以下能力：(1) 拮抗 RNA 与 A3G 的结合，以及 (2) 先发制人地激活细胞中的 A3G，从而抑制传入的 HIV 复制。在拟议的研究中，OyaGen, Inc 和 Sanford/Burnham 将合作：(1) 应用合理设计来合成三种经过验证的 A3G 选择性抗病毒化合物的化学修饰，(2) 选择具有增强活性的化合物作为 A3G:RNA 结合和抗病毒活性的拮抗剂，(3) 基于 A3G DNA 诱变活性和选择性的增强激活来选择最终探针细胞 A3G-RNA 复合物，以及。 (4) 根据体外 ADME/T 和小鼠体内 PK 对化合物进行分类，选择具有药物样特征的化合物。这些探针将带来重大的范式变化，因为它们将能够研究和监测 APOBEC3G 的作用机制，因为它在活病毒感染期间自然地在人类细胞中表达，并且将来在动物模型中也会表达。