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.

人类免疫缺陷病毒整合酶(IN)抑制剂正在被开发为潜在的抗艾滋病药物。虽然文献中已经报道了大量的抑制剂，但关于这些药物与IN蛋白的分子相互作用的信息很少。该项目的一个重点是设计和合成药理工具，以阐明抑制剂与IN的分子相互作用。在一项研究中，我们制备了含有联苯酮的香豆素作为光亲和配体。这些化合物与IN发生了交联化，通过质谱学鉴定了交联点，并通过诱变和分子模拟实验证实了交联点位于IN二聚体界面催化位置的远端。这些信息应该有助于设计在催化部位之外起作用的界面抑制剂。另一类重要的IN抑制剂是以芳基β-二酮酸为代表的，它们被认为是IN催化点内的金属螯合剂。为了阐明这些抑制剂与IN-DNA底物复合体的相互作用方式，在高亲和力的芳基β-二酮酸抑制剂上贴上了光亲和标记以及旨在促进光交联产物的分离和纯化的生物素标记。在其他研究中，HIV p6Gag蛋白与人Tsg101蛋白的结合已被证明是病毒萌发所必需的，并涉及p6蛋白的一个关键的9-聚“P-E-P-T-A-P-P-E-E”序列。我们正在准备作为Tsg101结合拮抗剂的9-聚体序列的多肽和多肽模拟变体，这可能会导致一类新的病毒萌发抑制物。一种方法是用N-取代甘氨酸(NSG)残基(称为“类肽”)取代Pro4残基。然而，这是一个综合问题。因此，我们求助于一类新的肽类变异体，这些变异体将肼类酰胺作为NSG的替代物。在固相树脂裂解后，通过将一系列醛与单一的高效液相纯化的酰肼前体反应，可以很容易地以库的方式制备这些化合物。这些肼类化合物被还原为N-取代的“类肽肼”，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。