Mechanism of Action of Novel Dual Actiing Pyrimidinediones

新型双作用嘧啶二酮类药物的作用机制

• 批准号: 7554994
• 负责人: Robert Walter Buckheit
• 金额: $ 27.25万
• 依托单位: IMQUEST BIOSCIENCES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-03 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7554994
• 关键词: Antiviral Agents; Applications Grants; Area; Binding; Binding Sites; Biochemical; Biological Assay; Cells; Clinical; Clinical Trials; Code; Combined Modality Therapy; Data; Discrimination; Dose; Drug resistance; Endopeptidases; Engineering; Enzymes; Epitopes; Evaluation; Exhibits; Gagging; Gene Combinations; Genes; Glycoproteins; Goals; HIV; HIV Envelope Protein gp120; HIV-1; HIV-2; Highly Active Antiretroviral Therapy; Human; In Vitro; Individual; Induced Mutation; Laboratories; Maps; Membrane; Methodology; Molecular Target; Monoclonal Antibodies; Multi-Drug Resistance; Mutate; Mutation; Patients; Pattern; Peptide Hydrolases; Pharmaceutical Preparations; Process; Protease Inhibitor; Public Health; Reproducibility; Research; Resistance; Reverse Transcription; Role; Salvage Therapy; Series; Site; Staging; Structure; Testing; Transmembrane Domain; Viral; Viral Genes; Viral Physiology; Virus; Virus Diseases; Virus Inhibitors; Virus Replication; design; drug resistant virus; env Gene Products; env Genes; established cell line; fitness; in vitro Assay; inhibitor/antagonist; non-nucleoside reverse transcriptase inhibitors; novel; pol genes; prevent; programs; recombinant virus; research study; small molecule; virus culture

DESCRIPTION (provided by applicant): The goal of this project is to define the mechanism of anti-HIV activity of the small molecule pyrimidinedione HIV inhibitor IQP-0410. IQP-0410 inhibits HIV replication by two distinct mechanisms, acting as a nonnucleoside reverse transcriptase inhibitor while also potently suppressing virus entry. Unlike other NNRTIs, IQP-0410 inhibits both HIV-1 and HIV-2. The ability of IQP-0410 to prevent HIV entry into target cells occurs through a unique mechanism and the compound has been found to be more potent against clinical strains of HIV than against laboratory-derived viruses. The proposed research will utilize in vitro assays for selection and genotypic and phenotypic characterization of viruses resistant to IQP-0410. The program will focus on resistance engendering mutations within the viral gag, pol and envelope genes. A series of chimeric viruses containing wild type HIV-1 and HIV-2 sequences and sequences from resistant viruses at early, mid, and late stages of the selection process will be engineered to examine the contribution of the individual domains to IQP-0410 resistance and fitness of the virus. Pseudotype viruses possessing envelope glycoproteins from wild type and IQP-0410-resistant HIV-1 and HIV-2 viruses will be evaluated for their ability to infect target cells in the presence and absence of IQP-0410 and to quantify the effects of env mutations on the antiviral activity of IQP-0410. Pseudotype viruses with site-specific mutations engineered into envelope gp120 and gp41 domains identified by resistance selection will be utilized to define the conformational epitope within the envelope structure targeted by IQP-0410 in entry inhibition. Additionally, experiments will be performed to further understand the role of mutations in the gag gene and evaluate potential effects of IQP-0410 on virus maturation from infected cells. The impact of resistance mutations on the activity of viral RT and protease will also be examined using biochemical analysis. PUBLIC HEALTH RELEVANCE The goal of this grant proposal is to define the mechanism of action of a small molecule inhibitor of human immunodeficiency virus (HIV). The HIV inhibitor (IQP-0410) is known to inhibit two essential stages of virus infection and replication (reverse transcription and virus entry), and thus has the potential for providing a combination therapy with administration of a single drug. The proposed research will define the molecular targets of the compound within the virus, the mechanism by which the compound inhibits virus replication, and mutations that occur in the virus that make it resistant to the compound and the effects of these mutations on virus replication and infectivity.

描述（由申请方提供）：本项目的目的是确定小分子嘧啶二酮HIV抑制剂IQP-0410的抗HIV活性机制。IQP-0410通过两种不同的机制抑制HIV复制，作为非核苷逆转录酶抑制剂，同时也有效抑制病毒进入。与其他NNRTI不同，IQP-0410可抑制HIV-1和HIV-2。IQP-0410阻止HIV进入靶细胞的能力是通过一种独特的机制发生的，并且已发现该化合物对HIV临床株比对实验室来源的病毒更有效。拟定研究将利用体外试验选择对IQP-0410耐药的病毒并进行基因型和表型表征。该计划将集中在耐药性产生的病毒gag，pol和包膜基因的突变。将对一系列含有野生型HIV-1和HIV-2序列以及筛选过程早期、中期和晚期耐药病毒序列的嵌合病毒进行工程改造，以检查单个结构域对IQP-0410耐药性和病毒适应性的贡献。将评价具有来自野生型和IQP-0410耐药HIV-1和HIV-2病毒的包膜糖蛋白的假型病毒在存在和不存在IQP-0410的情况下感染靶细胞的能力，并定量env突变对IQP-0410抗病毒活性的影响。将利用通过抗性选择鉴定的具有改造到包膜gp 120和gp 41结构域中的位点特异性突变的假型病毒来定义IQP-0410在进入抑制中靶向的包膜结构内的构象表位。此外，将进行实验以进一步了解gag基因突变的作用，并评价IQP-0410对感染细胞中病毒成熟的潜在影响。还将使用生化分析检查耐药突变对病毒RT和蛋白酶活性的影响。公共卫生相关性本拨款提案的目标是确定人类免疫缺陷病毒（HIV）小分子抑制剂的作用机制。已知HIV抑制剂（IQP-0410）抑制病毒感染和复制的两个基本阶段（逆转录和病毒进入），因此具有提供与施用单一药物的组合疗法的潜力。拟议的研究将确定病毒内化合物的分子靶点，化合物抑制病毒复制的机制，以及病毒中发生的使其对化合物产生抗性的突变以及这些突变对病毒复制和感染性的影响。