Intermolecular cooperativity in the action of antiviral agents

适当的抗逆转录病毒药物组合治疗可以减少HIV-1病毒血症， 低于检测限，并允许免疫系统的重建。的正确选择 抗逆转录病毒药物对于实现和维持病毒复制的抑制是至关重要的。 有趣的是，目前还没有一个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， 这取决于先前忽略的因子，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， 在这个实验室开发的一个新的指数中描述的，不同类别的抗逆转录病毒药物的差异， 超过10根原木（10，000，000，000倍！）以受斜率参数强烈影响的方式。因此 斜率参数是分析抑制潜力的关键缺失维度， 抗逆转录病毒药物这项提议旨在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， 抑制病毒复制。第一个具体的目的是测试一个机械假说，涉及一个独特的 一种分子间协同作用的形式，可以解释抗病毒药物剂量反应曲线的形状。 毒品这一假设将以一种非常具体的方式进行测试，使用表型混合实验， 携带药物靶向蛋白质的野生型和突变型的病毒。第二个目的 了解蛋白酶斜率参数的高幅度和药物间变异性 抑制剂类抗逆转录病毒药物。在迄今为止测试的药物类别中，蛋白酶抑制剂 显示最高斜率值。因此，这些药物中的一些的临床浓度可以抑制 单轮感染减少约10个对数。本项目旨在确定这一现象背后的机制， HIV-1对某些蛋白酶抑制剂的抑制作用的异常敏感性。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。由于高斜率值是 对于实现单轮感染的多对数抑制至关重要，所提出的分析可以识别 可能在体内特别有效的药物类别。第四个目标是确定效果 剂量-反应曲线斜率上的耐药突变。正确预测临床 耐药突变的后果需要了解突变的影响 斜率参数。最终目的是测量中和抗体的斜率参数 由于剂量-反应曲线斜率的概念也适用于HIV-1包膜蛋白， 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.