法布里病是由酶α-半乳糖苷酶A(GLA)缺乏引起的溶酶体贮积病。酶缺乏导致糖脂的积累,随着时间的推移,导致心血管疾病,脑血管,和肾脏疾病,最终导致生命的第四个或第五个十年死亡。目前,溶酶体贮积症通过酶替代疗法(ERT)通过对患者直接施用缺失的酶来治疗。鉴于它们作为药物递送系统的优势,脂质体越来越多地被研究和用于制药,食品和化妆品行业,但是市场的主要障碍之一是它们的可扩展性。将膨胀的液体有机溶液减压成水溶液(DELOS-susp)是一种基于压缩流体的方法,可实现具有显着的物理化学特性的纳米囊泡系统的可重复和可扩展的生产,在同质性方面,形态学,和颗粒大小。这项工作的目的是通过实施质量设计(QbD)方法来优化和达到体内临床前研究的合适配方,FDA和EMA推荐的开发强大的药物生产和控制方法的方法,制备用于治疗法布里病的载有α-半乳糖苷酶的纳米脂质体(nanoGLA)。通过风险分析和实验设计(DoE),我们获得了设计空间,其中发现GLA浓度和脂质浓度是实现稳定纳米制剂的关键参数。该设计空间允许优化该方法以生产适用于体内临床前测试的纳米制剂。
Fabry disease is a lysosomal storage disease arising from a deficiency of the enzyme α-galactosidase A (GLA). The enzyme deficiency results in an accumulation of glycolipids, which over time, leads to cardiovascular, cerebrovascular, and renal disease, ultimately leading to death in the fourth or fifth decade of life. Currently, lysosomal storage disorders are treated by enzyme replacement therapy (ERT) through the direct administration of the missing enzyme to the patients. In view of their advantages as drug delivery systems, liposomes are increasingly being researched and utilized in the pharmaceutical, food and cosmetic industries, but one of the main barriers to market is their scalability. Depressurization of an Expanded Liquid Organic Solution into aqueous solution (DELOS-susp) is a compressed fluid-based method that allows the reproducible and scalable production of nanovesicular systems with remarkable physicochemical characteristics, in terms of homogeneity, morphology, and particle size. The objective of this work was to optimize and reach a suitable formulation for in vivo preclinical studies by implementing a Quality by Design (QbD) approach, a methodology recommended by the FDA and the EMA to develop robust drug manufacturing and control methods, to the preparation of α-galactosidase-loaded nanoliposomes (nanoGLA) for the treatment of Fabry disease. Through a risk analysis and a Design of Experiments (DoE), we obtained the Design Space in which GLA concentration and lipid concentration were found as critical parameters for achieving a stable nanoformulation. This Design Space allowed the optimization of the process to produce a nanoformulation suitable for in vivo preclinical testing.