DDR SUBPRJ 5:FLEXIBLE PEPTIDE INHIBITORS INDUCING A STABLE CONFORMATION

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 HIV-1蛋白水解酶抑制剂是近几年发展起来的。然而，抗艾滋病毒药物治疗的一个主要困难是病毒基因组的快速突变，通过改变蛋白质靶标导致对药物的抗药性。联合疗法和鸡尾酒疗法已经开发出来，以应对病毒对蛋白酶抑制剂的抗药性。此外，许多耐药突变可能发生在远离活性部位的地方。这些观察结果支持一种不是每个抑制剂结构特有的耐药机制。相反，它们支持一种机制，该机制影响蛋白质酶关闭的动态过程和配体结合时的构象变化。这一应用的中心假设是，蛋白酶抑制剂应该考虑到配体结合时发生的动态过程。该方案中的抑制剂具有一个“还原”的多肽键和一个中间的苯丙氨酸基团，两端都有一个萘丙氨酸。我们试图通过检查这些多肽的核磁共振谱来确定它们的灵活性，并将这些抑制剂对接到HIV-1蛋白酶中进行计算分子动力学模拟，来验证我们的假设。一方面，有一定的灵活性是好的，因为多肽可以适应开放构象中酶的活性部位。另一方面，过于刚性的多肽抑制剂将减少最佳构象的数量，并将更不能适应影响其动态的蛋白酶的点突变。这项研究中积累的数据将为这项提议的长期目标奠定基础，即设计更有效的抑制剂，以抵抗艾滋病毒-1蛋白酶的突变。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Protease Inhibitors for HIV-1 protease have been developed in recent years. However, a major difficulty in the treatment with anti-HIV drugs has been the rapid mutations in the viral genome that result in resistance to the drug through changes in the protein target. Combination therapies and cocktails have developed in response to the resistance of the virus to protease inhibitors. Moreover, many resistant mutations can occur distant from the active site. These observations support a mechanism for drug resistance that is not specific to each inhibitor structure. Rather, they support a mechanism that affects a dynamic process of protease closure and conformational change upon ligand binding. The central hypothesis of this application is that the protease inhibitor should account for the dynamic process that occurs upon ligand binding. The inhibitors in this proposal have a "reduced" peptide bond and a phenylalanine group in the middle, and a naphthlylalanine on either end. We seek to prove our hypothesis through the specific aims of examining the NMR spectra of these peptides to determine their flexibility, and docking these inhibitors into HIV-1 protease performing computational molecular dynamic simulations. On the one hand, some flexibility is good in that the peptide can adapt to the active site of the enzyme in its open conformation. On the other hand, too rigid a peptide inhibitor will reduce the population of the best conformer and will be less able to adapt to the point mutations in the protease that affect its dynamics. The data accumulated in this investigation will lay the groundwork for the long range goals of this proposal to design more effective inhibitors that affect the dynamics of the HIV-1 protease in a manner that resist mutations in this enzyme.