Design and Synthesis of HIV Integrase as Potential Anti-

HIV整合酶（IN）抑制剂作为潜在的抗艾滋病药物正在被开发。尽管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。本项目的一个重点是设计和合成药理学工具来阐明抑制剂与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，并通过质谱鉴定了交联位点，并通过诱变和分子模拟实验证实了交联位点位于IN二聚体界面的催化位点的远端。这些信息将有助于设计在催化位点外起作用的界面抑制剂。另一类重要的IN抑制剂由芳基-二酮酸代表，被认为在IN催化位点内起金属螯合剂的作用。为了阐明这些抑制剂与In - dna底物复合物相互作用的方式，光亲和标签被附加到高亲和的芳基β -二酮酸抑制剂上，与生物素标签一起，旨在促进光交联产物的分离和纯化。抑制剂在in存在下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蛋白与人类Tsg101蛋白的结合已被证明是病毒出芽所必需的，并且涉及p6蛋白的一个关键的9-mer“p - e - p - p - t - a - p - p - e - e”序列。我们正在制备这种9-mer序列的肽和模拟肽变体，作为tsg101结合拮抗剂，这可能导致一类新的病毒出芽抑制剂。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，我们采用了一种新的肽类变体家族，其中包含腙酰胺作为NSG替代品。从固相树脂裂解后，通过一系列醛与单一hplc纯化的肼前体反应，可以很容易地以文库方式制备这些醛。将这些腙还原为n取代的“类肽肼”，为文库多样化提供了一条便捷的途径。这些研究促进了Tsg101结合抑制剂的设计。

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.