The Semen Enhancer of HIV Infection as a Novel Microbicide Target

项目总结/摘要 人类精液含有阳离子淀粉样纤维，称为“精液病毒感染增强剂”（SEVI）， 其强烈增强HIV-1感染，并且可能在病毒传播中起重要作用。我们的初步 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负责精液介导的HIV感染增强，以及（ii） SEVI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的创新方法， 到HIV-1杀微生物剂的开发，使用选择性靶向SEVI的药剂。高风险/高回报 这种方法与传统的针对病毒本身的杀微生物策略有着根本的不同， 预期与直接抗病毒方法高度互补。实际上，我们的长期目标是利用 SEVI靶向药物与传统杀微生物剂联合使用，以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 增强艾滋病毒感染。这种方法的可行性已经建立使用两个淀粉样蛋白结合 含有“屏蔽”低聚乙二醇（EG）部分的小分子：BTA-EG 4和-EG 6。这些 药物有效抑制SEVI和精液介导的HIV感染增强。在目标1中，我们将生成 并测试这些和其他淀粉样蛋白结合分子的新衍生物，包括寡价分子， 预期具有增加的SEVI结合亲和力。然后我们将测试他们抑制SEVI的能力， 使用一组R5病毒株（包括不同的进化枝和 传播菌株）。在目标2中，我们将研究新型淀粉样蛋白结合小分子之间的相互作用， 和女性生殖道的细胞。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通过暴露于HIV-1病毒体的HCEC增强PBMC的转感染。 R33阶段只有在实现明确的里程碑后才能进行。在目标3中，我们将使用 构效关系（SAR）数据，以完善我们的化学成分。我们还将测试我们的领先优势 分子在HIV-1感染的宫颈外植体模型中具有功效，以及它们是否具有协同或协同作用， 对其他候选杀微生物剂在精液存在下抑制HIV-1感染的能力的累加效应。 在最后的目标中，我们将评估最有希望的候选分子的毒性和炎症作用， 使用有益的乳杆菌菌株和宫颈外植体。R33阶段将以以下评估为高潮： 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.