The Semen Enhancer of HIV Infection as a Novel Microbicide Target

项目摘要/摘要 人类精液中含有阳离子淀粉样蛋白，被称为“病毒感染的精液增强剂”(SEVI)。 这有力地增强了HIV-1的感染，并可能在病毒传播中发挥重要作用。我们的预赛 数据显示，淀粉样蛋白结合分子与SEVI结合，并阻断精液介导的HIV-1增强 感染。这表明(I)SEVI对精液介导的HIV感染的增强负责，以及(Ii) 赛维代表了一种新的杀微生物剂靶标。因此，我们建议探索一种新颖、创新的方法 到艾滋病毒-1杀微生物剂的开发，使用选择性针对SEVI的试剂。这种高风险/高回报 这种方法从根本上不同于针对病毒本身的传统杀菌策略，而且是 预计将与直接抗病毒方法高度互补。事实上，我们的长期目标是利用 SEVI靶向剂与传统杀菌剂相结合，达到最佳的抗病毒效果。 在R21阶段，我们将测试新的淀粉样蛋白结合小分子是否抑制精液介导的 加强艾滋病毒感染。这种方法的可行性已经用两个淀粉样蛋白结合建立起来。 含有“屏蔽性”低聚乙二醇(EG)部分的小分子：BTA-EG4和-EG6。这些 药物有效地抑制SEVI和精液介导的艾滋病毒感染的增强。在目标1中，我们将生成 并测试这些分子和其他淀粉样蛋白结合分子的新衍生品，包括低价分子 预期具有更高的SEVI结合亲和力。然后我们将测试它们抑制SEVI的能力--以及 使用一组R5病毒株(包括不同的分支和 传播菌株)。在目标2中，我们将研究新的淀粉样蛋白结合小分子之间的相互作用。 以及来自女性生殖道的细胞。我们将评估我们的化合物是否对人类有毒 宫颈阴道上皮细胞(HCEC)，我们将测试它们是否抑制SEVI增强的HIV-1与 HCEC和/或SEVI增强暴露于HIV-1病毒粒子的HCEC对PBMC的反式感染。 只有在达到明确的里程碑时，才会进行R33阶段。在目标3中，我们将使用 结构-活性关系(SAR)数据，以提炼我们的化学成分。我们还将测试我们的领先优势是否 在HIV-1感染的宫颈移植模型中，分子具有有效性，无论它们是否具有协同作用或 在精液存在的情况下，对其他候选杀微生物剂抑制HIV-1感染能力的相加效应。 在最终目标中，我们将评估最有希望的候选分子的毒性和炎症效应， 使用有益的乳杆菌菌株和宫颈外植体。R33阶段将以评估 在兔阴道刺激(RVI)模型中最有希望的化合物的安全性和耐受性。 这些研究的总体目标是仔细确定靶向SEVI的小分子是否具有 作为一类新型杀微生物剂的潜在用途。

PROJECT SUMMARY/ABSTRACT Human semen contains cationic amyloid fibrils, termed the "Semen Enhancer of Virus Infection" (SEVI), which strongly enhance HIV-1 infection and may play an important role in viral transmission. Our preliminary data show that amyloid-binding molecules bind to SEVI, and block semen-mediated enhancement of HIV-1 infection. This suggests that (i) SEVI is responsible for semen-mediated enhancement of HIV infection and (ii) SEVI represents a novel microbicide target. We therefore propose to explore a novel, innovative approach to HIV-1 microbicide development, using agents that selectively target SEVI. This high-risk/high-reward approach is fundamentally different from traditional microbicidal strategies that target the virus itself, and is expected to be highly complementary with direct antiviral approaches. Indeed, our long-term goal is to use SEVI-targeting agents in combination with traditional microbicides, to achieve optimal antiviral effectiveness. In the R21 phase, we will test whether novel amyloid-binding small molecules inhibit semen-mediated enhancement of HIV infection. The feasibility of this approach has been established using two amyloid-binding small molecules which contain "shielding" oligo-ethylene glycol (EG) moieities: BTA-EG4 and -EG6. These agents efficiently inhibit SEVI- and semen-mediated enhancement of HIV infection. In Aim 1, we will generate and test novel derivatives of these and other amyloid-binding molecules, including oligovalent molecules that are expected to possess increased SEVI binding affinity. We will then test their ability to inhibit SEVI- and semen- mediated enhancement of HIV infection using a panel of R5 virus strains (including different clades and transmitted strains). In Aim 2, we will examine the interaction between novel amyloid-binding small molecules and cells from the female reproductive tract. We will evaluate whether our compounds are toxic to human cervicovaginal epithelial cells (HCEC), and we will test whether they inhibit SEVI-enhanced binding of HIV-1 to HCEC and/or SEVI-enhanced trans-infection of PBMC by HCEC exposed to HIV-1 virions. The R33 phase will be undertaken only if well-defined milestones are achieved. In Aim 3, we will use structure-activity relationship (SAR) data to refine our chemical compositions. We will also test whether our lead molecules have efficacy in a cervical explant model for HIV-1 infection, and whether they have a synergistic or additive effect on the ability of other candidate microbicides to inhibit HIV-1 infection in the presence of semen. In the final Aim, we will assess the toxicity and inflammatory effects of the most promising candidate molecules, using beneficial Lactobacilllus strains and cervical explants. The R33 phase will culminate with an evaluation of the safety and tolerability of the most promising compound in the rabbit vaginal irritation (RVI) model. The overall goal of these studies is to carefully determine whether small molecules that target SEVI have potential utility as a novel class of microbicides.