TRICIRIBINE AND NOVEL ANALOGS AS INHIBITORS OF HIV

三氯硝胺(TCN)及其水溶性前药(TCN-5‘-单磷酸， TCN-P)已被研究为抗癌药物；第二阶段临床试验 正在进行TCN-P。这两种化合物都不会被代谢成二恶英。 三磷酸，不结合到核酸中。最近我们 发现TCN和TCN-P具有抗人类免疫缺陷的活性 病毒(HIV)在急性感染和慢性感染的细胞中 纳摩尔浓度。这些浓度至少是一千- 比那些在未感染的人类细胞中具有细胞毒性的要低。 针对HIV-1和HIV-2临床小组的活动也得到了证明 分离株。对齐多夫定(AZT)和TIBO耐药的HIV对 TCN和TCN-P。分离的逆转录酶(RT)不受抑制 TCN或其核苷酸表明该化合物不通过 这个机制。 基于这些新发现，这项提议的广泛目标是 以TCN为先导化合物，发明具有更大容量的新试剂 艾滋病毒的活性和选择性。一种系统的化学程序 合成新的TCN类似物旨在开发新的化合物 将对其进行抗艾滋病毒活性和细胞毒性评估 人类细胞。再加上对行动模式的详细审查 针对HIV的TCN，这些研究应该允许设计和合成 比母体更活跃和/或毒性更小的化合物 化合物，并可能阐明艾滋病毒治疗的新靶点。 具体目标包括合成TCN的新的同系物，包括 但不限于：(I)杂环取代类似物，(Ii) 糖类似物，(Iii)环修饰化合物，和(Iv)C-核苷。 化学合成的方向将由抗病毒药物和 新化合物的细胞毒性评价和作用模式研究。 评估将包括对急性感染患者进行合胞菌斑分析。 单元格和以下一种或多种方法，并使用acutally和 慢性感染的T细胞、U1细胞或外周血白细胞 (PBL)：感染性病毒产量减少、p24核心抗原或RT检测。 TCN和活性类似物也将结合现有的 抗病毒药物用于确定是否增强了抗HIV-1的活性 在没有伴随细胞毒性增强的情况下发生。数据将是 使用三维剂量-反应方法进行分析，并在 统计上显著的协同作用或拮抗作用将被识别出来 量化的。

Triciribine (TCN) and its water soluble prodrug (TCN-5'-monophosphate, TCN-P) have been studied as anticancer drugs; phase II clinical trials are in progress with TCN-P. Neither compound is metabolized to the di- or triphosphate and is not incorporated into nucleic acids. Recently we discovered that TCN and TCN-P are active against human immunodeficiency virus (HIV) in acutely infected and chronically infected cells at nanomolar concentrations. These concentrations are at least a thousand- fold lower than those which are cytotoxic in uninfected human cells. Activity also was demonstrated against a panel of HIV-1 and HIV-2 clinical isolates. HIV resistant to zidovudine (AZT) and TIBO remained sensitive to TCN and TCN-P. Isolated reverse transcriptase (RT) was not inhibited by TCN nor its nucleotides demonstrating that the compound does not act via this mechanism. Based upon these new discoveries, the broad objective of this proposal is to use TCN as a lead compound and invent new agents having greater activity and selectivity for HIV. A systematic program of chemical synthesis of new TCN analogs has been designed to develop novel compounds which will be evaluated for activity against HIV and for cytotoxicity in human cells. Coupled with a detailed examination of the mode of action of TCN against HIV, these studies should permit the design and synthesis of compounds which are more active and/or less toxic than the parent compounds and may elucidate a new target for HIV therapy. The specific aims involve the synthesis of new congeners of TCN including but not limited to the following: (i)heterocycle-substituted analogs, (ii) sugar analogs, (iii) ring-modified compounds, and (iv) C-nucleosides. Direction for the chemical synthesis will be provided by antiviral and cytotoxicity evaluation of the new compounds and mode of action studies. Evaluations will include a syncytial plaque assay in acutely infected cells and one or more of the following methods using acutely and chronically infected T-cells, U1-cells, or peripheral blood leukocytes (PBL): infectious virus yield reduction, p24 core antigen, or RT assays. TCN and active analogs also will be studied in combination with existing antiviral drugs to determine if potentiation of activity against HIV- 1 occurs without a concomitant potentiation of cytotoxicity. Data will be analyzed using three-dimensional dose-response methodology and areas of statistically significant synergy or antagonism will be identified and quantitated.