Design and development of a novel, thermostable, and inhalable dry powder COVID-19 vaccine

摘要 SARS冠状病毒2(SARS-CoV-2)是一种高度传染性的病毒 已知会导致2019年冠状病毒病(新冠肺炎)。这种疾病已被宣布为一种 世界卫生组织的全球大流行。在美国，截至2021年1月20日， 我们有2400多万感染者，其中约40万人死亡。新冠肺炎 在患有高血压和糖尿病等并存疾病的患者中似乎更致命 临床表现多样，如肺炎、腹泻、多器官功能衰竭等。 在被感染的人中。此外，预计感染人数和死亡率将会增加。 根据IHME模型，未来几个月将大幅增长。有一个紧急的全球问题 需要开发一种耐热性和自我管理的疫苗。表面的尖峰蛋白 冠状病毒是开发疫苗的极佳候选者，目前已在 EUA获得了疫苗，因为它在病毒进入宿主细胞方面发挥着重要作用。 脂质体是一种很好的药物递送系统，它可以将抗原以颗粒形式呈现在 自然，并允许加入一种佐剂，帮助产生强大的免疫 回应。此外，脂质体可以设计成与病毒的大小相似，并且 SPEKE蛋白可以偶联到脂质体表面，以模拟自然呈现在 病毒。免疫细胞会像病毒一样内化和处理抗原，并产生 中和抗体滴度。目前市场上的大多数疫苗都需要冷链 (冷藏)储存和分发疫苗以维持效力，并要求访问 诊所由训练有素的医务人员进行免疫接种。这些是实现以下目标的瓶颈 大流行中的大规模疫苗接种进一步增加了医院和供应链的压力。在……里面 此外，一个主要限制是现有制造设施的制造能力有限。 数十亿剂。在这项提案中，我们的目标是开发一种耐热和可吸入的干粉 疫苗易于扩大规模，不需要冷链储存和运输，而且 个人在家中通过简单的吸入即可自行给药。为了实现这一目标，我们将 追求两个目标：1)研制可吸入、耐热的干粉新冠肺炎疫苗 含有S蛋白吸附的脂质体载体和2)中和粘膜和 新冠肺炎干粉疫苗雾化免疫后系统免疫球蛋白抗体滴度的测定 在老鼠身上。这一项目的顺利完成将对 干粉新冠肺炎疫苗，特别是在探索耐热和可吸入疫苗方面 易于制造、储存和分发，这些特征对大流行至关重要。

Summary Severe Acute Respiratory Syndrome-Coronavirus 2 (SARS-CoV-2) is a highly infectious virus known to cause the 2019 coronavirus disease (COVID-19). The disease has been declared as a global pandemic by the World Health Organization. In the United States, as of January 20, 2021, we have more than 24 million infected individuals with approximately 400,000 deaths. COVID-19 appears to be more deadly in patients with co-morbidities such as hypertension and diabetes and presents many clinical manifestations such as pneumonia, diarrhea, multiple organ failure, etc. among those infected. Moreover, the number of infections and fatality rate is expected to increase significantly over the next few months, according to the IHME model. There is an urgent global need to develop a thermostable and self-administrable vaccine. The spike protein on the surface of coronavirus is an excellent candidate for developing vaccines, has been used in the currently EUA obtained vaccines, as it plays an important role in the entry of the virus into the host cell. Liposomes are excellent drug delivery systems that can present the antigen as particulate in nature and allow the incorporation of an adjuvant that aids in the generation of a robust immune response. Further, the liposomes can be designed to be of similar size to that of the virus and the spike protein can be conjugated to the liposomal surface to mimic the natural presentation on the virus. The immune cells will internalize and process the antigen like the virus and generate neutralizing antibody titers. Most of the vaccines currently in the market require cold chain (refrigeration) to store and distribute the vaccine to maintain the efficacy and require a visit to the clinic for immunization by trained medical personnel. These serve as bottlenecks in achieving mass vaccination in a pandemic adding further stress to the hospitals and the supply chain. In addition, a major limitation is the limited ability of existing manufacturing facilities to manufacture billions of doses. In this proposal, we aim to develop a thermostable and inhalable dry powder vaccine that is easy to scale-up, eliminates the need for cold-chain storage and transport, and is self-administrable by individuals’ at-home by simple inhalation. To achieve this goal, we will pursue two aims: 1) Formulation of an inhalable and thermostable dry powder COVID-19 vaccine containing S protein-adsorbed liposomal carriers and 2) Evaluation of neutralizing mucosal and systemic IgA and IgG antibody titers after aerosol administration of dry powder COVID-19 vaccine in mice. Successful completion of this project will have a profound impact on the development of dry powder COVID-19 vaccine, particularly in exploring thermostable and inhalable vaccines that are easy to manufacture, store, and distribute, characteristics that are critical in pandemic.