Intermolecular cooperativity in the action of antiviral agents

使用适当的抗逆转录病毒药物组合治疗可将HIV-1病毒血症降低到 低于检测极限，并允许重建免疫系统。正确的选择 抗逆转录病毒药物对于实现和维持对病毒复制的抑制至关重要。 有趣的是，没有被广泛接受的系统来比较不同药物的抗病毒活性。 该实验室以前的研究表明，抗逆转录病毒药物的抑制潜力很强。 这取决于以前被忽略的因素，这里称为斜率参数。此参数 描述了剂量-反应曲线的陡度。抗逆转录病毒药物的抑制潜力， 在本实验室开发的新指数中描述的，不同类别的抗逆转录病毒药物 超过10个对数(10,000,000,000倍！)以一种受坡度参数强烈影响的方式。因此， 斜率参数是抑制势分析中的一个关键缺失维度 抗逆转录病毒药物。这项提议试图了解这一现象背后的分子机制。 并利用这些信息指导药物和疫苗的开发，这些药物和疫苗将最大限度地 抑制病毒复制。第一个特定的目标是测试一个涉及唯一的 可解释抗病毒药物剂量-效应曲线形状的分子间协作性形式 毒品。这一假设将使用表型混合实验以非常具体的方式进行验证 携带药物靶标蛋白的野生型和突变型的病毒。第二个目标 是为了了解酶的斜率参数的高幅度和药物间的可变性 抑制类抗逆转录病毒药物。到目前为止，在测试的药物类别中，蛋白酶抑制剂 显示最高的坡度值。因此，其中一些药物的临床浓度可以抑制 单轮感染约10个对数。这个项目试图确定这一现象背后的机制 HIV-1对某些蛋白水解酶抑制剂的特殊敏感性。第三个具体目标是 将这种分析方法应用于新型抗逆转录病毒药物。因为高斜率值是 对于实现单轮感染的多对数抑制至关重要，拟议的分析可以确定 在体内可能特别有效的药物类别。第四个目标是确定效果 耐药突变在剂量-反应曲线斜率上的变化。正确的临床预测 耐药突变的后果需要了解突变的影响 在坡度参数上。最终目的是测量中和抗体的斜率参数。 针对HIV-1被膜蛋白，因为剂量-反应曲线斜率的概念也适用于 疫苗诱导的效应机制，包括中和抗体。

Treatment with appropriate combinations of antiretroviral drugs can reduce HIV-1 viremia to below the limit of detection and allow reconstitution of the immune system. The correct choice of antiretroviral drugs is critical for achieving and maintaining suppression of viral replication. Interestingly, there is no widely accepted system for comparing the antiviral activity of different drugs. Previous studies from this lab have shown that the inhibitory potential of antiretroviral drugs is strongly dependent upon a previously ignored factor termed here the slope parameter. This parameter describes the steepness of the dose-response curve. The inhibitory potential of antiretroviral drugs, described in a new index developed by this lab, varies for different classes of antiretroviral drugs by over 10 logs (10,000,000,000 fold!) in a manner that is strongly influence by the slope parameter. Thus the slope parameter is a critical missing dimension in the analysis of the suppressive potential of antiretroviral drugs. This proposal seeks to understand the molecular mechanisms underlying this effect and to use that information to guide the development of drugs and vaccines that will maximally suppress viral replication. The first specific aim is to test a mechanistic hypothesis involving a unique form of intermolecular cooperativity which can explain the shapes of dose-response curves for antiviral drugs. This hypothesis will be tested in a very specific way using phenotypic mixing experiments with viruses carrying both wild type and mutant forms of the proteins targeted by the drugs. The second aim is to understand the high magnitude and drug-to-drug variability of the slope parameter for the protease inhibitor class of antiretroviral drugs. Among the drug classes tested to date, the protease inhibitors show the highest slope values. As a result, clinical concentrations of some of these drugs can inhibit single round infections by ~10 logs. This project seeks to identify the mechanism underlying this extraordinary susceptibility of HIV-1 to inhibition by some protease inhibitors. The third specific aim is to apply this method of analysis to novel classes of antiretroviral drugs. Since a high slope value is critical for achieving multi-log inhibition of single round infection, the proposed analyses could identify drug classes that are likely to be particularly effective in vivo. The fourth aim is to determine the effect of drug resistance mutations on dose-response curve slope. Correct prediction of the clinical consequences of drug resistance mutations requires an understanding of the effects of the mutations on the slope parameter. The final aim is to measure the slope parameter of neutralizing antibodies directed at the HIV-1 envelope protein since concept of dose-response curve slope also applies to vaccine-induced effector mechanisms including neutralizing antibodies.