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）：仅限于逆转录酶和蛋白酶抑制剂的联合疗法显着改善了 HIV 感染患者的预后。尽管取得了初步成功，但对 FDA 批准的多种抗逆转录病毒药物产生耐药性的 HIV 毒株的流行率不断增加，要求设计针对 HIV 新治疗靶点（例如包膜蛋白）的药物。 HIV 感染是通过病毒包膜与细胞膜上的 CD4 结合而引发的。这会诱导 gp120 发生构象变化，从而揭示在病毒进入过程中重要的隐藏表位。特别是，gp120 V3 环与趋化因子受体的相互作用发生在 gp120 构象变化之后，此时 V3 环变得更加暴露。 V3 环通过与趋化因子受体 CCR5 或 CXCR4 相互作用而成为 HIV 趋向性的关键决定因素，抗 V3 环单克隆抗体可以中和。 VIN2 环可能是屏蔽 V3 环和其他与适当的 HIV 共受体结合所必需的区域的重要决定因素。由于 VIN2 和 V3 结构域包含辅助受体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点。 我们的目标是鉴定针对可能抑制病毒进入的 VIN2 和 V3 区域的高亲和力小肽。我们计划生成包含这些环的杂合蛋白，用作从组合文库中选择特定高亲和力肽的靶标。为了获得这些环的特异性肽，我们将生成限制 gp120 V3 或 VlN2 环移植到鼠白血病病毒 (MLV) 包膜蛋白或葡萄球菌核酸酶 (SN) 区域的构建体，这些结构是充分表征的支架蛋白，可以耐受大插入，同时保持其天然结构。噬菌体展示将用于从组合文库中选择针对这些环的高亲和力肽。然后将测定噬菌体衍生肽对V1N2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，针对包膜结构的肽已在对其他抗逆转录病毒疗法产生耐药性的患者中证明了临床疗效。由于我们的肽将针对病毒结构，因此我们期望它们具有低毒性，并且与当前的抗逆转录病毒药物没有交叉耐药性。