Rational Design of HIV Entry Inhibitors

HIV进入抑制剂的合理设计

• 批准号: 6799431
• 负责人: Monique Regail Ferguson
• 金额: $ 22.27万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6799431
• 关键词: HIV envelope protein gp120; Staphylococcus; chimeric proteins; computer simulation; drug design /synthesis /production; drug screening /evaluation; inhibitor /antagonist; murine leukemia virus; nuclease; peptide library; phage display; protein binding; protein protein interaction; protein structure function; surface plasmon resonance; virus infection mechanism

DESCRIPTION (provided by applicant): Combination therapy, restricted to reverse transcriptase and protease inhibitors, has dramatically improved outcomes of HIV-infected patients. In spite of the initial success, the increased prevalence of HIV strains resistant to multiple FDA-approved antiretroviral drugs mandates the design of agents directed to new therapeutic targets in HIV, such as the envelope protein. HIV infection is initiated by binding of the viral envelope to CD4 at the cell membrane. This induces a conformational change in gp120 to reveal cryptic epitopes important in viral entry. In particular, interaction of gp120 V3 loop with chemokine receptors occurs after gp120 cenformational changes, when the V3 loop becomes more exposed. The V3 loop is a critical determinant for HIV tropism via its interaction with the chemokine receptor CCR5 or CXCR4, and anti-V3 loop monoclonal antibodies can be neutralizing. The VIN2 loop may be an important determinant in shielding the V3 loop and other regions essential for binding to the appropriate HIV co-receptor. Since the VIN2 and V3 domains contain critical determinants for coreceptor interactions, these loops are suitable targets for the design of small molecule inhibitors. Our goal is to identify high-affinity small peptides that target VIN2 and V3 regions that may inhibit viral entry. We plan to generate hybrid proteins containing these loops for use as targets in the selection of specific high affinity peptides from combinatorial libraries. In order to obtain peptides specific to these loops, we will generate constructs that constrain gp120 V3 or VlN2 loops grafted into regions of murine leukemia virus (MLV) envelope protein or Staphylococcal nuclease (SN), which are well-characterized scaffold proteins that can tolerate large insertions while maintaining their native structure. Phage display will be used to select high affinity peptides directed to these loops from combinatorial libraries. The affinity of phagederived peptide(s) for the VlN2 and V3 constructs will then be assayed. There is high probability that small molecules binding to these gp120 regions will disrupt viral entry, and may be used for the development of new antiretroviral drugs. Currently, peptides targeting the envelope structure have demonstrated clinical efficacy in patients resistant to other antiretroviral therapies. Since our peptides will be targeting viral structures, we expect them to have low toxicity, and no cross-resistance with the current antiretroviral drugs.

描述（由申请人提供）：联合治疗，仅限于逆转录酶和蛋白酶抑制剂，已大大改善了感染HIV的患者的预后。尽管取得了最初的成功，但抗多种FDA批准的抗逆转录病毒药物的HIV菌株的患病率增加，这授权针对HIV新治疗靶标的剂的设计，例如包膜蛋白。 HIV感染是通过病毒包膜与细胞膜CD4结合而引发的。这引起了GP120的构象变化，以揭示在病毒进入中重要的神秘表位。特别是，当V3回路变得更加暴露时，GP120 V3环与趋化因子受体的相互作用发生在GP120增长变化后。 V3环是HIV对tropism的关键决定因素，它与趋化因子受体CCR5或CXCR4相互作用，抗V3环路单克隆抗体可以中和。 VIN2回路可能是屏蔽V3环和与适当HIV共受体所必需的其他区域的重要决定因素。由于VIN2和V3结构域包含对共感受器相互作用的关键决定因素，因此这些环是设计小分子抑制剂的合适靶标。 我们的目标是确定靶向可能抑制病毒式进入的Vin2和V3区域的高亲和力小肽。我们计划生成包含这些环的混合蛋白，以用作组合文库的特定高亲和肽的靶标。为了获得特定于这些环的肽，我们将生成构造的构建体，以限制接枝到鼠白血病病毒（MLV）包膜蛋白或葡萄球菌核酸酶（SN）的区域的gp120 v3或VLN2环，这些构造可以耐受耐受性的网核蛋白（SN）大型插入，同时保持其本地结构。噬菌体显示器将用于选择与组合库的这些环的高亲和力肽。然后将分析含肽对VLN2和V3构建体的亲和力。与这些GP120区域结合的小分子很可能会破坏病毒的进入，并且可用于开发新的抗逆转录病毒药物。目前，针对包膜结构的肽表现出对其他抗逆转录病毒疗法具有抗性患者的临床功效。由于我们的肽将靶向病毒结构，因此我们预计它们的毒性低，并且与当前的抗逆转录病毒药物没有交叉抗性。