Design and Synthesis of HIV Integrase as Potential Anti-

HIV整合酶（IN）抑制剂正被开发为潜在的抗艾滋病药物。目前正在研究的一类先导结构可以广泛地表征为芳基β-二酮家族。已独立报道该类成员在胞外酶测定中表现出对HIV整合酶的有效抑制，并在HIV感染的细胞中提供良好的抗病毒作用。通过系统地设计和合成大量的芳基β-二酮类似物，我们已经开发了新的含叠氮基的芳基β-二酮变体，其在细胞外测定中表现出高的IN抑制效力，并在HIV感染的细胞中提供具有降低的细胞毒性的抗病毒效果细胞。最近的工作集中在用萘啶药效团替换我们的含叠氮基的抑制剂的β-二酮部分，这在其他临床相关的HIV-1整合酶抑制剂中显示出实用性。为了阐明这些和其他抑制剂与IN DNA底物复合物相互作用的方式，化学和光活化亲和标记已被掺入高亲和力抑制剂中。我们还开发了一种新型的药理学工具的第一批成员，该工具通过IN酶中氨基酸残基的位点特异性乙酰化作用作为“亲和乙酰化剂”发挥作用。目前正在进行质谱研究，以阐明与酶孵育后这些试剂的共价连接位点。最近的研究已经检查了与其他烷基化官能团相比，这些亲和乙酰化剂对各种氨基酸的亲核侧链的反应性和选择性。基于微分反应性，一类新的？双功能？正在开发亲和配体，其可适用于广泛的蛋白质-配体相互作用的研究。在单独的研究中，正在进行合作，以获得与HIV整合酶结合的抑制剂的X射线结构。从这种X射线结构中获得的信息应该为计算机辅助设计有效的新抑制剂提供一个起点。

Inhibitors of HIV integrase (IN) are being developed as potential anti-AIDS drugs. One class of lead structure currently under investigation can be broadly characterized as being of the aryl beta-diketo family. Members of this class have been reported independently to exhibit potent inhibition of HIV integrase in extracellular enzyme assays and to provide good antiviral effects in HIV-infected cells. Through the systematic design and synthesis of a large number of aryl beta-diketo analogues, we have developed novel azido containing aryl beta-diketo variants, which exhibit high IN inhibitory potency in extracellular assays and provide antiviral effects cells with reduced cytotoxicity in HIV infected. Recent work has focused on replacement of the beta-diketo portion of our azido containing inhibitors with the naphthyridine pharmacophore, which has shown utility in other clinically-relevant HIV-1 integrase inhibitors. In order to elucidate the manner in which these and other inhibitors interact with IN DNA substrate complexes, chemical and photo-activatable affinity labels have been incorporated into high affinity inhibitors. We have also developed the first members of a novel, new class of pharmacological tool that function as "affinity acetylators" by site-specific acetylation of amino acid residues in the IN enzyme. Mass spectral studies are currently ongoing to elucidate sites of covalent attachment by these agents following incubation with the enzyme. Recent studies have examined the reactivities and selectivities towards the nucleophilic side chains of various amino acids of these affinity acetylators as compared with other alkylating functionalities. Based on differential reactivities, a new class of ?bifunctional? affinity ligands is being developed that may have applicability in the study of a broad range protein-ligand interactions. In separate studies, collaborative efforts are underway to obtain X ray structures of inhibitors bound to the HIV integrase enzyme. Information obtained from such X-ray structures should provide a starting point for the computer-assisted design of potent new inhibitors.