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.