High-throughput Computational Docking Screen for Small-molecule Inhibitors of HIV

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过程中，药物毒性和耐药分离株的出现要求寻找可用于开发新型抗病毒药物的新靶点。HIV-1基质蛋白(MA)是一种结构蛋白，参与逆转录病毒生命周期的几个阶段。尽管MA在病毒粒子组装中起着至关重要的作用，但有关该功能的细节，以及负责调节该功能的结构域，仍在不断涌现。MA还参与了HIV-1基因的核输入，最近还参与了感染过程中的新角色，包括病毒进入/去涂层，细胞骨架介导的运输，以及将病毒组装到脂筏上。MA在HIV-1感染中的重要性及其作为抗病毒靶点的潜力已经被两种旨在阻断其功能的策略的成功所证明，一种是基于体内的，另一种是最近使用小分子的方法。鉴于这些研究的成功和HIV-1 MA的广泛功能，我们使用了高通量计算对接(HTCD)方法来识别可能潜在地与HIV-1 MA结合并破坏其功能的小分子。从这个HTCD屏幕上，我们已经确定了19个小分子，它们被预测在结构和功能上重要的区域与HIV-1 MA结合。从这19个化合物中，我们确定了8个化合物具有抗病毒活性。重要的是，表现出抗病毒活性的化合物可以分为三类：早期抑制剂、晚期抑制剂和两种似乎同时干扰早期和晚期事件的化合物。据我们所知，这些化合物是第一批小分子抑制剂，可以破坏HIV-1复制周期的早期和晚期过程。在这项建议中，我们希望通过研究它们的结合亲和力、化学计量比和MA上的结合位置(特定目标1)，以及确定它们的范围、特异性和作用机制(特定目标2)，建立适合于通过药物化学进行进一步优化的MA靶向化合物。我们相信，这一跨学科战略的实施将成功地找到药理上有用的试剂，这些试剂可以用来深入了解艾滋病毒-1MA蛋白的生物学功能，特别是它在艾滋病毒-1生命周期早期整合前阶段的作用(S)。此外，从这项研究中发现的小分子有望成为一种新型抗病毒药物的先导化合物。 与公共卫生相关：尽管抗逆转录病毒药物疗法取得了成功，但抑制艾滋病毒-1复制的新目标是非常可取的，需要新的药物来应对日益严重的耐药性问题。本申请中提出的研究将表征一类以HIV-1基质(MA)蛋白为目标的化合物，这些化合物在HIV-1生命周期的多个阶段通过一种新的机制发挥作用。该提案中确定的化合物有可能作为新型抗艾滋病毒药物的模板，同时也提供了分析艾滋病毒-1基本生物学的试剂。

DESCRIPTION (provided by applicant): Despite recent progress in anti-HIV therapy, drug toxicity and the 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 antiviral agents. The HIV-1 matrix protein (MA) is a structural protein involved in several stages in the life cycle of the retrovirus. Although MA has long been known to be crucial for virion assembly, details regarding this function, and the domains responsible for mediating it, are still emerging. MA has also been implicated in nuclear import of HIV-1 cDNA and has recently been implicated in novel roles during infection including viral entry/uncoating, cytoskeletal-mediated transport, and targeting viral assembly to lipid rafts. The importance of MA in HIV-1 infection and its potential as an antiviral target have been demonstrated by the success of two strategies designed to block its function, one intrabody-based and another, more recent approach using small molecules. Given the success of these studies and the breadth of functions of HIV-1 MA, we have used a high-throughput computational docking (HTCD) approach to identify small molecules that may potentially bind to and disrupt the functions of HIV-1 MA. From this HTCD screen, we have identified 19 small molecules that are predicted to bind to HIV-1 MA in a structurally and functionally important region. From these 19 compounds, we have determined that eight possess antiviral activity. Importantly, the compounds that display antiviral activity can be grouped into three classes: early-stage inhibitors, late-stage inhibitors, and two compounds that appear to disrupt both early and late events. To our knowledge, these compounds represent the first small-molecule inhibitors that can disrupt both early- and late-stage processes in the replication cycle of HIV-1. In this proposal, we wish to establish the MA-targeted compounds that are suitable for further optimization via medicinal chemistry by investigating their binding affinity, stoichiometry, and binding site on MA (Specific Aim 1) in addition to determining their range, specificity, and mechanism of action (Specific Aim 2). We believe that execution of this cross-disciplinary strategy will successfully identify pharmacologically useful reagents that can be used to gain insight into the biological functions of HIV-1 MA protein, especially its role(s) in the early, pre-integration stage of the HIV-1 life cycle. Moreover, small molecules identified from this study hold promise as lead compounds for a new class of antiviral agent. PUBLIC HEALTH RELEVANCE: Despite the success of antiretroviral drug regimens, new targets for inhibition of HIV-1 replication are highly desirable, and new drugs are needed to combat the rising problem of drug resistance. The studies proposed in this application will characterize a class of compounds targeted at the HIV-1 matrix (MA) protein and that act via a novel mechanism at multiple stages in the HIV-1 life cycle. The compounds identified within this proposal have the potential to serve as templates for novel anti-HIV agents while also providing reagents with which to dissect the basic biology of HIV-1.