Oxadaizols: Rationally designed compounds targeting HIV nuclear importation

描述(申请人提供)：尽管最近在抗艾滋病毒治疗方面取得了进展，但在艾滋病毒感染者的长期治疗中，药物毒性和耐药分离株的出现要求寻找新的靶点，以用于开发新的抗病毒药物。其中一个目标是艾滋病毒-1整合前复合体的核移位过程。在我们的初步研究中，我们鉴定了一类恶二唑化合物，它们通过靶向基质蛋白(MA)来抑制HIV-1核进口。这些化合物是通过计算机辅助药物设计来选择的，并被预测在其核定位信号(NLS)附近与MA结合，从而阻止MA介导的核进口。该组的先导化合物ITI-367在T淋巴细胞和巨噬细胞的培养中显示出强大的抗HIV活性，并在体外培养的淋巴组织中抑制HIV-1的复制。实时定量聚合酶链式反应分析表明，ITI-367特异性抑制病毒DNA的核输入。在这一应用中，我们建议进一步表征ITI-367的分子作用机制，并将这一分析扩展到与ITI-367相关的其他化合物，试图确定最有效的HIV-1复制抑制剂。为了实现这些目标，将实现以下具体目标：1.生产和鉴定对恶二唑化合物具有抗药性的艾滋病毒-1变异体。2.表征恶二唑类化合物与MA的相互作用，并分析其抗HIV活性。这一探索性建议完全符合这一PA的目标，因为它解决了艾滋病毒研究中的一个创新概念，这对基础和临床研究都有影响。拟议的实验有望为关于这类新的抗艾滋病毒化合物作用机制的假设提供直接测试。完成后，这些研究有望确定有效的抗艾滋病毒化合物通过一种不同于任何其他目前使用的药物的新机制发挥作用。在这项申请中提出的研究将表征一类化合物，目标是艾滋病毒生命周期中的一个新靶点：将病毒DNA运输到细胞核。这些研究与公共卫生高度相关，因为它们有望扩大现有抗艾滋病毒药物的范围，从而为抗击艾滋病毒感染者常见的耐药性提供机会。

DESCRIPTION (provided by applicant): Despite recent progress in anti-HIV therapy, drug toxicity and emergence of drug-resistant isolates during long- term treatment of HIV-infected patients necessitate the search for new targets that can be used to develop novel anti-viral agents. One such target is the process of nuclear translocation of the HIV-1 pre-integration complex. In our preliminary studies, we identified a class of oxadiazol compounds that inhibit HIV-1 nuclear import by targeting the matrix protein (MA). These compounds were selected using computer-assisted drug design and are predicted to bind to MA near its nuclear localization signal (NLS) thus blocking MA-mediated nuclear import. The lead compound from this group, ITI-367, showed potent anti-HIV activity in cultures of T lymphocytes and macrophages, and also inhibited HIV-1 replication in ex vivo cultured lymphoid tissue. Real- time PCR analysis demonstrated that ITI-367 specifically inhibited nuclear import of viral DNA. In this application, we propose to further characterize the molecular mechanism of action of ITI-367 and to expand this analysis to other compounds related to ITI-367 in an attempt to identify the most potent inhibitor of HIV-1 replication. Towards these goals, the following Specific Aims will be pursued: 1. To produce and characterize the HIV-1 variants resistant to oxadiazol compounds. 2. To characterize interaction between oxadiazol derivatives and MA and analyze their anti-HIV activity. This exploratory proposal is fully consistent with the goals of this PA as it addresses an innovative concept in HIV research which has implications both for basic and clinical studies. Proposed experiments are expected to provide a direct test for the hypothesis regarding the mechanism of action of this new class of anti-HIV compounds. Upon completion, these studies are expected to define potent anti-HIV compounds working through a novel mechanism different from that of any other currently used drug. Studies proposed in this application will characterize a class of compounds aimed at a novel target in HIV life cycle: transport of the viral DNA to the nucleus. These studies are highly relevant to public health as they are expected to expand the repertoire of available anti-HIV drugs, thus providing an opportunity to fight drug resistance often seen in HIV-infected patients.