Improved Nanoparticle Targeting of Tissue Myeloid Cells for HIV-1 Long-acting Pre-exposure Prophylaxis

摘要 抗逆转录病毒暴露前预防(PrEP)是防止病毒幼稚传播的重要工具 在目前结束艾滋病毒流行的努力中，它发挥着重要的作用。如果每天服用目前的口服PrEP 这些策略可以可靠地阻止艾滋病毒的传播。但是，严格遵守每日服药、服药的要求 疲劳和其他体制障碍使口腔PrEP得不到充分利用。长效注射用PrEP 策略有可能解决与口头PrEP相关的许多问题，但实现 确保药物持续释放至少三个月的药物配方和给药策略 仍然具有挑战性，并推动了全新的长效PrEP战略的发展。这个项目 开发一种基于膜包裹纳米粒(NPs)的长效可注射PrEP策略 维持抗逆转录病毒药物(ARV)在原代组织中抑制浓度的细胞库 艾滋病毒-1在女性生殖道(FGT)和直肠的传播地点。CD169的选择性靶向表达 巨噬细胞和树突状细胞是通过神经节苷脂GM3掺入神经节苷脂完成的。 薄膜。GM3-CD169结合触发NPs在非内溶酶体隔室的摄取和隔离 与组织相关巨噬细胞中的病毒包含间隔(VCC)有明显的相似之处 树突状细胞。这些隔间代表了受保护的地方，NPs可以从那里将药物释放到 在较长时间内包围组织。膜包裹的嵌段共聚物反胶束将 作为长效PrEP的GM3-NP平台。长效替诺福韦(TFV)和 恩曲他滨(FTC)前药将被用作活性化合物来验证该方法。嵌段共聚物 NPS将包含TFV，该TFV共价连接到包裹持有FTC的水核的聚合物外壳 与树枝状大分子偶联。在体外定量载药和释药后，GM3介导的靶向 将对人源化小鼠模型FGT和直肠中的CD169髓系细胞进行测试。同时，GM3- 将对NP平台进行优化，以实现药物在靶组织的持续释放。这一假设认为 优化的NPs在人源化小鼠模型中提供了至少三年的粘膜HIV感染保护 几个月将受到考验。本申请的具体目标是： 目的1：制备用于TFV/FTC缓释的膜包裹多组分纳米粒。 目的：靶向CD169在FGT和SLT中表达的髓系细胞，以获得TFV/FTC的持续释放。 目的3：证实TFV/FTC对人源化小鼠粘膜传播HIV-1的长期保护作用 加入GM3-NPs。

Summary Antiretroviral pre-exposure prophylaxis (PrEP) is an important tool for preventing transmission to virus naïve individuals and plays an important role in current efforts to end the HIV epidemic. If taken daily current oral PrEP strategies reliably block HIV transmission. However, the requirement of strict adherence to daily pill uptake, pill fatigue and other institutional barriers to access leave oral PrEP underutilized. Long-acting injectable PrEP strategies have the potential to address many of the problems associated with oral PrEP but the realization of drug formulations and delivery strategies that ensure sustained drug release for at least three months has remained challenging and motivates the development of entirely new long-acting PrEP strategies. This project develops a long-acting injectable PrEP strategy based on membrane-wrapped nanoparticles (NPs) that establish cellular depots for sustained maintenance of inhibitory concentrations of antiretrovirals (ARVs) at primary tissue sites of HIV-1 transmission in the female genital tract (FGT) and rectum. Selective targeting of CD169-expressing macrophages and dendritic cells is accomplished through incorporation of the ganglioside GM3 in the NP membrane. GM3-CD169 binding triggers uptake and sequestration of NPs in non-endolysosomal compartments that share distinct similarities with virus containing compartments (VCCs) in tissue-associated macrophages and dendritic cells. These compartments represent protected sites from where NPs can release drugs into the surrounding tissue for an extended period of time. Membrane-wrapped inverse micelles of block copolymers will be engineered as a GM3-NP platform for long-acting PrEP. A combination of long-acting tenofovir (TFV) and emtricitabine (FTC) prodrugs will be used as active compounds to validate the approach. The block copolymer NPs will contain TFV covalently linked to a polymer shell that encapsulates an aqueous core holding FTC conjugated to dendrimers. After quantifying drug loading and release in vitro, the GM3-mediated targeting of CD169+ myeloid cells in the FGT and rectum of a humanized mouse model will be tested. In parallel, the GM3- NP platform will be optimized to achieve sustained drug release in the target tissues. The hypothesis that the optimized NPs provide protection from mucosal HIV infection in a humanized mouse model for at least three months will be tested. The specific aims of this application are: Aim 1: To develop membrane-wrapped multicomponent NPs for sustained release of TFV/FTC. Aim 2: To target CD169-expressing myeloid cells in the FGT and SLTs for sustained TFV/FTC release. Aim 3: To demonstrate long-term protection from mucosal HIV-1 transmission in humanized mice by TFV/FTC incorporating GM3-NPs.