Intermolecular cooperativity in the action of antiviral agents

描述(由申请人提供)：使用适当的抗逆转录病毒药物组合治疗可以将艾滋病毒-1病毒血症减少到检测到的限度以下，并允许重建免疫系统。正确选择抗逆转录病毒药物对于实现和保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强烈依赖于一个以前被忽视的因素，在这里称为斜率参数。该参数描述了剂量-反应曲线的陡度。在本实验室开发的一项新指数中描述的抗逆转录病毒药物的抑制潜力，对于不同类别的抗逆转录病毒药物，其抑制潜力的差异超过10个对数(10,000,000倍！)以一种受坡度参数强烈影响的方式。因此，斜率参数是抗逆转录病毒药物抑制潜力分析中的一个关键缺失维度。这项提议试图了解这种效应背后的分子机制，并利用这些信息来指导将最大限度地抑制病毒复制的药物和疫苗的开发。第一个具体目标是测试一种机制假说，该假说涉及一种独特的分子间协同性，它可以解释抗病毒药物的剂量-反应曲线的形状。这一假设将以一种非常具体的方式进行验证，方法是使用携带野生型和突变型蛋白质的病毒进行表型混合实验，这些病毒是药物的靶标。第二个目的是了解抗逆转录病毒药物的蛋白酶抑制剂类药物的斜率参数的高幅度和药物间的可变性。在迄今测试的药物类别中，蛋白酶抑制剂显示出最高的斜率值。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。该项目试图确定HIV-1对某些蛋白酶抑制剂抑制的这种特殊敏感性背后的机制。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。由于高斜率值对于实现对单轮感染的多对数抑制至关重要，建议的分析可以确定在体内可能特别有效的药物类别。第四个目的是确定耐药突变对剂量-反应曲线斜率的影响。正确预测耐药突变的临床后果需要了解突变对斜率参数的影响。最终目的是测量针对HIV-1包膜蛋白的中和抗体的斜率参数，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。公共卫生相关性：该项目涉及一种在实验室中测量艾滋病毒药物抗病毒活性的新方法。该方法包含了一个以前未被认识到的因素，它极大地促进了药物和疫苗抑制艾滋病毒的能力。其目标是促进最大限度地抑制艾滋病毒的药物和疫苗的设计。

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: This project involves a new method for measuring the antiviral activity of HIV drugs in the laboratory. The method incorporates a previously unappreciated factor that contributes enormously to the ability of drugs and vaccines to inhibit HIV. The goal is to facilitate the design of drugs and vaccines that maximally inhibit HIV.