Villa-Hermosilla Monica-Carolina, Negro Sofia, Barcia Emilia, Hurtado Carolina, Montejo Consuelo, Alonso Mario, Fernandez-Carballido Ana
Department of Pharmaceutics and Food Technology, School of Pharmacy, Universidad Complutense de Madrid, Plaza de Ramón y Cajal s/n, 28040 Madrid, Spain.
Institute of Industrial Pharmacy, School of Pharmacy, Universidad Complutense de Madrid, Plaza de Ramón y Cajal s/n, 28040 Madrid, Spain.
Pharmaceutics. 2022 Jun 30;14(7):1392. doi: 10.3390/pharmaceutics14071392.
Inhalation therapy is gaining increasing attention for the delivery of drugs destined to treat respiratory disorders associated with cytokine storms, such as COVID-19. The pathogenesis of COVID-19 includes an inflammatory storm with the release of cytokines from macrophages, which may be treated with anti-inflammatory drugs as celecoxib (CXB). For this, CXB-loaded PLGA microparticles (MPs) for inhaled therapy and that are able to be internalized by alveolar macrophages, were developed. MPs were prepared with 5% and 10% initial percentages of CXB (MP-C1 and MP-C2). For both systems, the mean particle size was around 5 µm, which was adequate for macrophage uptake, and the mean encapsulation efficiency was >89%. The in vitro release of CXB was prolonged for more than 40 and 70 days, respectively. The uptake of fluorescein-loaded PLGA MPs by the RAW 264.7 macrophage cell line was evidenced by flow cytometry, fluorescence microscopy and confocal microscopy. CXB-loaded PLGA MPs did not produce cytotoxicity at the concentrations assayed. The anti-inflammatory activity of CXB (encapsulated and in solution) was evaluated by determining the IL-1, IL-6 and TNF-α levels at 24 h and 72 h in RAW 264.7 macrophages, resulting in a higher degree of reduction in the expression of inflammatory mediators for CXB in solution. A potent degree of gene expression reduction was obtained with the developed CXB-loaded MPs.
吸入疗法在用于递送旨在治疗与细胞因子风暴相关的呼吸系统疾病（如 COVID-19）的药物方面正受到越来越多的关注。COVID-19 的发病机制包括巨噬细胞释放细胞因子引发的炎症风暴，可用塞来昔布（CXB）等抗炎药物进行治疗。为此，开发了用于吸入治疗且能够被肺泡巨噬细胞内化的载 CXB 的聚乳酸-羟基乙酸共聚物（PLGA）微粒（MPs）。MPs 分别以 5%和 10%的初始 CXB 百分比制备（MP-C1 和 MP-C2）。对于这两种体系，平均粒径约为 5 µm，这足以被巨噬细胞摄取，平均包封率>89%。CXB 的体外释放分别延长了 40 多天和 70 多天。通过流式细胞术、荧光显微镜和共聚焦显微镜证实了 RAW 264.7 巨噬细胞系对载荧光素的 PLGA MPs 的摄取。在所测定的浓度下，载 CXB 的 PLGA MPs 未产生细胞毒性。通过测定 RAW 264.7 巨噬细胞在 24 小时和 72 小时时的白细胞介素-1（IL-1）、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）水平，评估了 CXB（包封的和溶液中的）的抗炎活性，结果表明溶液中的 CXB 对炎症介质表达的降低程度更高。所开发的载 CXB 的 MPs 实现了显著程度的基因表达降低。
