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

项目总结 提交此竞争性修订申请是为了扩大我们正在进行的NIH的范围 授予UH3HL141797以利用我们的人体单芯片器官(器官芯片)微流控培养设备 用于快速开发和评估潜在的新冠肺炎治疗药物。我们正在进行的UH3 格兰特支持开发人肺芯片作为快速发现新的新的体外临床前工具 由流感引起的病毒性大流行的治疗。在最近的研究中，我们发现高度分化的 我们肺部芯片中的人类细胞，以及我们开发的肠道芯片中的人类肠道细胞，表达 高水平的ACE2和TMPRSS2，介导SARS-CoV-2病毒(CoV2)感染。我们还能够 用表达CoV2尖峰蛋白的病毒伪颗粒(CoV2pp)感染这些器官芯片 天然CoV2病毒在与FDA批准的多种药物进行细胞检测时的效果。 人类肺芯片也被证明是评估潜在的COVID19抑制作用的更严格的模型 仅作为这些药物的子集的活性在给药时显著抑制CoV2pp的进入 临床研究报告了其在人体血液中的最大浓度(Cmax)的芯片上流动。在这里，我们 建议将人肠和肺芯片与计算发现和合成相结合 开发广谱冠状病毒疗法的化学方法 现在有19名患者，并使我们能够做好准备，防止感染相关的大流行病毒 出现在未来。在初步研究中，用我们的计算工具设计了多种新型化合物 在细胞内对CoV2pp和天然CoV2病毒进行测试时，显示出显著的抑制活性 化验。因此，我们的具体目标包括：1)使用计算和合成化学方法来 产生预计可抑制CoV2病毒和相关冠状病毒感染的新化合物，2) 在基于细胞的检测中，通过分析活性分子的结构-活性关系来确定活性分子的优先顺序 天然CoV2和相关冠状病毒，3)识别基于抑制的先导化合物和有效剂量 使用本地冠状病毒的人体器官芯片中的感染和宿主炎症反应，以及4) 在小鼠体内进行药代动力学研究，并结合迭代化学合成和基于细胞的测试 测试以优化先导化合物的药用性能和安全性，同时保持有效性。 通过这项工作，我们将发现具有广谱抑制活性的新化合物。 对抗CoV2以及相关冠状病毒，并产生必要的药代动力学数据来移动这些 药物进入动物验证研究，并最终进入人体临床试验。这项工作也将进一步确立 人体器官芯片作为临床前工具加速药物开发的价值。

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.