Lung-on-a-Chip Disease Models for Efficacy Testing (COVID-19 Competitive Revision)

项目摘要 这一竞争性修订申请正在提交，以扩大我们正在进行的NIH的范围， 授权UH 3 HL 141797，以利用我们的人体器官芯片（器官芯片）微流控培养装置 用于快速开发和评估COVID-19的潜在治疗药物。我们正在进行的UH 3 赠款支持开发人肺芯片作为体外临床前工具，用于快速发现新的 用于治疗由流感引起的病毒大流行。在最近的研究中，我们发现高分化的 我们的肺芯片中的人类细胞，以及我们开发的肠芯片中的人类肠细胞， 高水平的ACE 2和TMPRSS 2介导SARS-CoV-2病毒（CoV 2）感染。我们还能够 用表达CoV 2刺突蛋白的病毒假颗粒（CoV 2 pp）感染这些器官芯片， 天然CoV 2病毒在基于细胞的试验中针对多种FDA批准的药物进行测试时的作用。 人肺芯片也被证明是用于评估潜在COVID 19抑制的更严格的模型。 活性，因为这些药物中只有一个子集在给药时显著抑制CoV 2 pp的进入。 在临床研究中报告的人血液中的最大浓度（Cmax）下的芯片上流动。这里我们 我建议使用人类肠道和肺芯片结合计算发现和合成 化学方法来开发广谱冠状病毒疗法， COVID 19患者现在，并允许我们做好准备，以防止感染相关的大流行病毒， 出现在未来。在初步研究中，使用我们的计算工具设计了多种新型化合物， 当在基于细胞的细胞培养中针对CoV 2 pp和天然CoV 2病毒测试时， 测定。因此，我们的具体目标包括：1）使用计算和合成化学方法， 创造新的化合物，预计可抑制CoV 2病毒和相关冠状病毒的感染，2） 通过在基于细胞的检测中分析活性分子的结构-活性关系， 天然CoV 2和相关冠状病毒，3）基于抑制作用确定先导化合物和有效剂量 使用天然冠状病毒的人器官芯片中的感染和宿主炎症反应，以及4） 在小鼠中进行药代动力学研究，结合迭代化学合成和基于细胞的测试， 分析以优化先导化合物的药物特性和安全性，同时保持功效。 通过这项工作，我们将确定新的化合物，表现出广谱抑制活性 针对CoV 2以及相关冠状病毒，并生成必要的药代动力学数据， 药物进入动物验证研究，最终进入人体临床试验。这项工作还将进一步建立 人体器官芯片作为加速药物开发的临床前工具的价值。

PROJECT SUMMARY This COMPETITIVE REVISION application is being submitted to expand the scope of our ongoing NIH grant UH3HL141797 in order to leverage our human organ-on-a-chip (Organ Chip) microfluidic culture devices for the rapid development and assessment of potential therapeutic agents for COVID-19. Our ongoing UH3 grant supports the development of human Lung Chips as in vitro preclinical tools for rapid discovery of new therapeutics for viral pandemics caused by influenza. In recent studies, we showed that highly differentiated human cells in our Lung Chips, as well as human intestinal cells within Intestine Chips we developed, express high levels of ACE2 and TMPRSS2 that mediate SARS-CoV-2 virus (CoV2) infection. We also were able to infect these Organ Chips with CoV2 spike protein-expressing viral pseudoparticles (CoV2pp) that closely mimic the effects of native CoV2 virus when tested against multiple FDA approved drugs in cell-based assays. Human Lung Chips were also shown to be more stringent models for assessing potential COVID19 inhibitory activity as only a subset of these drugs significantly inhibited entry of the CoV2pp when administered under flow on-chip at their maximum concentration (Cmax) in human blood reported in clinical studies. Here, we propose to use human Intestine and Lung Chips in combination with computational discovery and synthetic chemistry approaches to develop broad-spectrum coronavirus therapeutics that would both help infected COVID19 patients now, and allow us to be prepared to prevent infections by related pandemic viruses that emerge in the future. In preliminary studies, multiple novel compounds designed with our computational tools exhibited significant inhibitory activities when tested against both CoV2pp and native CoV2 virus in cell based assays. Thus, our Specific Aims include: 1) to use computational and synthetic chemistry approaches to create new compounds that are predicted to inhibit infection by CoV2 virus and related coronaviruses, 2) to prioritize active molecules by analyzing their structure-activity relationships in cell-based assays infected with native CoV2 and related coronaviruses, 3) to identify lead compounds and effective doses based on inhibition of infection and host inflammatory responses in human Organ Chips using native coronaviruses, and 4) to carry out pharmacokinetic studies in mice coupled with iterative chemical synthesis and testing in cell-based assays to optimize the pharmaceutical properties and safety of the lead compounds, while retaining efficacy. Through this effort, we will identify new compounds that demonstrate broad spectrum inhibiting activities against CoV2 as well as related coronaviruses, and generate pharmacokinetic data necessary to move these drugs into animal validation studies and, eventually, human clinical trials. This work will also further establish the value of human Organ Chips as preclinical tools for accelerating drug development.