Viral and Host Proteins as Therapeutic Targets

病毒和宿主蛋白作为治疗靶点

• 批准号: 8349026
• 负责人: Stuart F. J. Le Grice
• 金额: $ 78.98万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8349026
• 关键词: Address; Affect; Antiviral Agents; Attention; Binding; Binding Sites; Child Development; Child health care; Clinical; Collaborations; DNA-Directed DNA Polymerase; Data; Development; Enzymes; Excision; Fluorescence Polarization; Goals; HIV; HIV Drug Resistance Program; HIV-1; HIV-2; Human; Inhibitory Concentration 50; Institutes; Integrase; Juniperus occidentalis; Legal patent; Libraries; Methods; Molecular Target; Nucleosides; Proteins; RNA-Directed DNA Polymerase; Reporting; Reverse Transcriptase Inhibitors; Ribonuclease H; Robotics; Screening procedure; Site; Site Visit; Tropolone; Universities; Urea; Viral; Virus Replication; Western Red Cedar; base; calanolide A; inhibitor/antagonist; lens epithelium-derived growth factor; novel; protein protein interaction; small molecule; therapeutic target

A multicenter collaboration between the NCI-Frederick Molecular Targets Discovery and HIV Drug Resistance Programs, the National Institute of Child Health and Development, and the University of Pittsburgh has used high-throughput robotics to screen several libraries, totaling 250,000 compounds, for small-molecule inhibitors of HIV RNase H function. Secondary screening against bacterial and human RNase H has addressed whether selectivity for the retroviral enzyme can be achieved. Several structural classes of RNase H inhibitors have been identified by this strategy, the most potent of which was the hydroxylated tropolone beta-thujaplicinol. Derived from the bark of the western cedar Thuja plicata, beta-thujaplicinol inhibited HIV-1 RT/RNaseH at a concentration of 0.2 uM, while the IC50 for human RNase H was 6.0 uM and that of the bacterial enzyme >50 uM. In addition, beta-thujaplicinol was shown to synergize with the nonnucleoside inhibitor calanolide A, strengthening contentions from other groups that both the DNA polymerase and RNase H activities of HIV-1 RT can be simultaneously targeted. Vinylogous ureas constitute a second structural class of RNase H inhibitors, and a patent covering these inhibitors has been submitted. Structural studies to define the binding site of the most potent RNase H inhibitors are currently underway. We are continuing our studies on RNase H as an antiviral target by (1) using crystallographic data to alter residues of RT implicated in inhibitor binding, (2) synthesizing novel derivatives of both structural classes, and (3) investigating the relationship between impaired RNase H function and increased excision of chain-terminating nucleoside RT inhibitors (NRTIs). Site-specific derivatization with a novel trifunctional agent will also be investigated as a general method of creating fluorescent proteins, allowing fluorescence polarization to be used for screening protein:protein interactions. Initial studies will focus on the interaction of the host protein lens epithelium-derived growth factor (LEDGF) with HIV-1 integrase. [Corresponds to Le Grice Project 3 in the April 2007 site visit report of the HIV Drug Resistance Program]

NCI-Frederick分子目标发现与抗HIV耐药性计划，国家儿童健康与发展研究所以及匹兹堡大学之间的多中心合作使用了HIV HIV RNase HIV RNase HIV RNase H函数的小分子抑制剂的高通用机器人。针对细菌和人类RNase H的次级筛查已经解决了是否可以实现逆转录病毒酶的选择性。该策略已经确定了RNase H抑制剂的几种结构类别类别，其中最有效的是羟基化的Troponβ-硫代克环醇。 beta-thujaplicinol源自西雪松Thuja plicata的树皮，以0.2 um抑制HIV-1 RT/RNASEH，而人RNase H的IC50为6.0 um，细菌酶> 50 um的IC50 um。此外，显示β-硫代克林醇与非核苷抑制剂calanolide A协同作用，从而加强了其他群体的观点，即HIV-1 RT的DNA聚合酶和RNase H活性都可以同时靶向。乙烯基尿素构成了RNase H抑制剂的第二个结构类别，并且已提交了覆盖这些抑制剂的专利。目前正在进行定义最有效的RNase H抑制剂的结合位点的结构研究。我们正在使用（1）使用晶体学数据继续对RNase H作为抗病毒靶点进行研究，以改变与抑制剂结合有关的RT残基，（（2）合成两种结构类别的新型衍生物，以及（3）研究损伤RNase H功能与链末期型核苷型核苷RT rt Inibisitors（NRTIS）的关系之间的关系。与新型三功能剂的位点特异性衍生化也将作为产生荧光蛋白的一般方法研究，从而使荧光极化可用于筛选蛋白质：蛋白质相互作用。最初的研究将集中于宿主蛋白透镜上皮衍生的生长因子（LEDGF）与HIV-1积分酶的相互作用。 [在2007年4月的艾滋病毒耐药性计划的现场访问报告中，对应于Le Grice项目3]