Pharmacology of HIV Viral DNA & Retroviral Integrases

HIV病毒DNA的药理学

• 批准号: 6950193
• 负责人: YVES POMMIER
• 金额: --
• 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/6950193
• 关键词: antiAIDS agent; antiviral agents; binding sites; cofactor; crosslink; drug design /synthesis /production; drug interactions; drug screening /evaluation; enzyme inhibitors; human immunodeficiency virus 1; integrase; intermolecular interaction; ketoacid; pharmacokinetics; protein structure function; virus DNA; virus infection mechanism; virus replication

In an effort to further extend the number of targets for development of anti-retroviral agents, we are studying HIV-1 integrase inhibitors using in vitro assays using recombinant enzyme and short oligonucleotides corresponding to the proviral ends (LTR's). Integrase is a rationale target for inhibitor development because it is essential for viral replication. It is encoded by the viral genome and does not have a cellular equivalent. Our laboratory has pioneered this research field and reported various families of inhibitors. We have written several reviews on this topic; the most recent will be published in early 2004. This year, the first class of HIV-1 integrase inhibitors has been introduced in clinical trials. We are investigating these diketo acid (DKA) derivatives in collaboration with Dr. Terrence Burke (Laboratory of Medicinal Chemistry, CCR, NCI) and Dr. Vinay Pathak (Antiviral Drug Resistance Program, CCR, NCI). We have elucidated the structure-activity relationship for this new type of compounds, and found that DKAs discriminate between wild-type and mutant HIV-1 integrase and between magnesium and manganese, the two metal cofactors for integrase. Our goal is to elucidate the drug binding site(s) in the enzyme-DNA complex, and to discover agents with a greater therapeutic index and/or novel structural motifs. We discovered that azido derivatives of diketo acids are potent and selective anti-integrase inhibitors, and are antiviral. The azido group can chelate the divalent metal in the enzyme catalytic site. A patent application has been filed for these derivatives. We are also studying novel types of inhibitors that can prevent integration by binding to the proviral DNA ends as well as small peptide and nucleotide inhibitors. The peptide inhibitors are also antiviral and have been patented. We are also studying the molecular interactions between integrase and its DNA substrate using enzyme-DNA crosslinking assays in order to model drug-enzyme-DNA interactions. A study using minor groove scanning with benzo[a]pyrene diol epoxide dG-N2 adduct is in press in The Journal of Biological Chemistry. We recently found a novel interaction between one of the viral DNA bases and the enzyme.

为了进一步扩大抗逆转录病毒药物的靶标数量，我们使用重组酶和对应于前病毒末端（LTR）的重组酶和短寡核苷酸（LTR）研究了HIV-1积分酶抑制剂。整合酶是抑制剂开发的基本原理目标，因为它对于病毒复制至关重要。它由病毒基因组编码，没有细胞等效。我们的实验室开创了这一研究领域，并报告了各种抑制剂家族。我们已经写了几篇有关此主题的评论。最新的将于2004年初出版。今年，在临床试验中引入了第一类HIV-1整合酶抑制剂。我们正在与Terrence Burke博士（CCR，NCI）和Vinay Pathak博士（抗病毒耐药性计划，CCR，NCI）合作研究这些二甲酸（DKA）衍生物。我们已经阐明了这种新型化合物的结构活性关系，并发现DKA会区分野生型和突变的HIV-1积分酶以及镁和锰（镁和锰），镁和锰是两种集成酶的金属辅因子。我们的目标是阐明酶-DNA复合物中的药物结合位点，并发现具有更大治疗指数和/或新型结构基序的药物。我们发现，二酮酸的氮化衍生物是有效的抗积酶抑制剂，并且是抗病毒。偶氮基团可以在酶催化位点螯合二价金属。这些衍生工具已提交专利申请。我们还正在研究新型的抑制剂类型，这些抑制剂可以通过与前病毒DNA末端以及小肽和核苷酸抑制剂结合来防止整合。肽抑制剂也是抗病毒，已获得专利。我们还使用酶-DNA交联测定法研究了整合酶及其DNA底物之间的分子相互作用，以模拟药物 - 酶-DNA相互作用。在生物化学杂志上使用苯并[A] pyrene Diol环氧化物DG-N2加合物进行少量凹槽扫描的研究。最近，我们发现了一个病毒DNA碱基和酶之间的新型相互作用。