Abstract：
One of the strategies proposed for the neutralization of SARS-CoV-2 has been to synthetize small proteins able to act as a decoy towards the virus spike protein, preventing it from entering the host cells. In this work, the incorporation of one of these proteins, LCB1, within a spray-dried formulation for inhalation was investigated. A design of experiments approach was applied to investigate the optimal condition for the manufacturing of an inhalable powder. The lead formulation, containing 6% w/w of LCB1 as well as trehalose and L-leucine as excipients, preserved the physical stability of the protein and its ability to neutralize the virus. In addition, the powder had a fine particle fraction of 58.6% and a very high extra-fine particle fraction (31.3%) which could allow a peripheral deposition in the lung. The in vivo administration of the LCB1 inhalation powder showed no significant difference in the pharmacokinetic from the liquid formulation, indicating the rapid dissolution of the microparticles and the protein capability to translocate into the plasma. Moreover, LCB1 in plasma samples still maintained the ability to neutralize the virus. In conclusion, the optimized spray drying conditions allowed to obtain an inhalation powder able to preserve the protein biological activity, rendering it suitable for a systemic prevention of the viral infection via pulmonary administration.
摘要：
提出的中和SARS-CoV-2的策略之一是合成能够充当病毒刺突蛋白诱饵的小蛋白，阻止它进入宿主细胞。在这项工作中,掺入这些蛋白质中的一种，研究了用于吸入的喷雾干燥制剂中的LCB1。实验方法的设计被用来研究制造可吸入粉末的最佳条件。铅配方，含有6％w/w的LCB1以及海藻糖和L-亮氨酸作为赋形剂，保留了蛋白质的物理稳定性及其中和病毒的能力。此外,粉末具有58.6%的细颗粒分数和非常高的超细颗粒分数(31.3%),这可以允许在肺中的外周沉积。粉末的体内给药与液体制剂的药代动力学没有显着差异，表明微粒的快速溶解及其转移到血浆中的能力。此外，血浆样品中的LCB1仍然保持中和病毒的能力。总之,优化的喷雾干燥条件允许获得能够保持蛋白质生物活性的吸入粉末，使其适于通过肺部给药系统预防病毒感染。
