Design and Synthesis of HIV Integrase as Potential Anti-

作为潜在抗病毒药物的 HIV 整合酶的设计和合成

• 批准号: 7337944
• 负责人: TERRENCE BURKE
• 金额: --
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7337944
• 关键词: Design; Synthesis; HIV; Integrase; Potential

Inhibitors of HIV integrase (IN) are being developed as potential anti-AIDS drugs. Although a large number of inhibitors have been reported in the literature, little information has been forthcoming regarding the molecular interactions of these agents with IN protein. One focus of this project is to design and synthesize pharmacological tools to clarify molecular interactions of inhibitors with IN. In one study we prepared biphenyl ketone-containing coumarins as photoaffinity ligands. These were cross-linked to IN and the site of cross-linking was identified by mass spectroscopy and confirmed by mutagenesis and molecular modeling experiments to be distal to the catalytic site at the IN dimer interface. This information should aid in the design of interfacial inhibitors that act outside of the catalytic site. Another important class of IN inhibitors is represented by the aryl beta-diketo acids that are thought to function as metal chelators within the IN catalytic site. In order to elucidate the manner in which these inhibitors interact with IN-DNA substrate complexes, photoaffinity labels were appended onto high affinity aryl beta-diketo acid inhibitors along with biotin tags intended to facilitate isolation and purification of photo-cross-linked products. Photoactivation studies of inhibitors in the presence of IN along with MALDI-TOF mass spectral identification of cross-linked products is ongoing.In other studies, binding of the HIV p6Gag protein to human Tsg101 protein has been shown to be necessary for viral budding and to involve a critical 9-mer "P-E-P-T-A-P-P-E-E" sequence of the p6 protein. We are preparing peptide and peptide mimetic variants of this 9-mer sequence as Tsg101-binding antagonists that may lead to a new class of viral budding inhibitors. One approach was to replace the Pro4 residue with N-substitued glycine (NSG) residues (termed "peptoids"). However, this is synthetically problematic. therefore, we resorted to a new family of peptoid variants that incorporate hydrazone amides as NSG surrogates. These can be preparede readily in library fashion by reacting a series of aldehydes with a single HPLC-purified hydrazide precursor following cleavage from the solid-phase resin. Reduction of these hydrazones to N-substitued "peptoid hydrazides" affords a facile route to library diversification. These studies are advancing the design of Tsg101 binding inhibitors.

HIV整合酶（IN）抑制剂正在被开发为潜在的抗艾滋病药物。尽管文献中报道了大量抑制剂，但关于这些药物与 IN 蛋白的分子相互作用的信息很少。该项目的重点之一是设计和合成药理学工具，以阐明抑制剂与 IN 的分子相互作用。在一项研究中，我们制备了含联苯酮的香豆素作为光亲和配体。它们与 IN 交联，交联位点通过质谱鉴定，并通过诱变和分子建模实验确认交联位点位于 IN 二聚体界面催化位点的远端。这些信息应该有助于设计在催化位点之外起作用的界面抑制剂。另一类重要的 IN 抑制剂以芳基 β-二酮酸为代表，它们被认为在 IN 催化位点内充当金属螯合剂。为了阐明这些抑制剂与 IN-DNA 底物复合物相互作用的方式，将光亲和标签与生物素标签一起附加到高亲和力芳基 β-二酮酸抑制剂上，旨在促进光交联产物的分离和纯化。 IN 存在下抑制剂的光活化研究以及交联产物的 MALDI-TOF 质谱鉴定正在进行中。在其他研究中，HIV p6Gag 蛋白与人 Tsg101 蛋白的结合已被证明对于病毒出芽和涉及 p6 蛋白的关键 9 聚体“P-E-P-T-A-P-P-E-E”序列。我们正在制备该 9 聚体序列的肽和肽模拟变体作为 Tsg101 结合拮抗剂，这可能会产生一类新的病毒出芽抑制剂。一种方法是用 N-取代的甘氨酸 (NSG) 残基（称为“类肽”）替换 Pro4 残基。然而，这综合起来是有问题的。因此，我们采用了一个新的类肽变体家族，其中包含腙酰胺作为 NSG 替代物。这些可以通过一系列醛与单一 HPLC 纯化的酰肼前体反应，然后从固相树脂裂解，以文库方式轻松制备。将这些腙还原为 N-取代的“类肽酰肼”为文库多样化提供了一条简便的途径。这些研究正在推进 Tsg101 结合抑制剂的设计。