• stabilized reverse micelle
• nanomedicine
• drug delivery
• Krabbe disease
• nervous central system
• nanovectors
• Angiopep-2
• GALC

Krabbe disease (KD) is an autosomal recessive lysosomal storage disorder (LSD) caused by deficiency of the galactosylceramidase (GALC) enzyme. In humans, KD is typically a neurodegenerative disease of early infancy. No effective cure is currently available for KD.
The most clinically employed approach to treat LSDs is enzyme replacement therapy (ERT), that is done by systematically administering the missing/defective enzyme. However, this direct approach has many limitations; in fact, it is not effective for lysosomal storage diseases affecting the central nervous system (CNS), like KD, due to the presence of the Blood Brain Barrier (BBB).
A promising strategy to improve biodistribution and efficacy of many molecules is the encapsulation in a nanovector.
In a recent study, a system based on polymeric reverse micelles (RMs) has been developed for the internalization of hydrophilic drugs. Unlike classic micelles, RMs have the peculiarity of having a hydrophilic core consisting of PEG (polyethylene glycol) and an outer portion consisting of PLGA (poly(lactide-co-glycolide)). Moreover, this protocol has led to the formation of micelles stabilized by a chemical crosslink. All these features make RMs particularly suitable for serving as nanocarriers for hydrophilic compounds.
In this thesis, stabilized RMs were functionalized with peptide Angiopep-2 (RMs-Ang2), which is already known for its capability to penetrate the BBB. These engineered RMs were loaded with GALC with the aim to facilitate the targeted delivery of the enzyme to the CNS and act as potential nanodrug for systemic enzyme replacement therapy in KD.
Control RMs, RMs-Ang2 and RMs-Ang2 loaded with GALC were synthetized and characterized for: i. size and surface potential by Dynamic Light Scattering (DLS); ii. enzyme loading and release capabilities by quantifying protein content and enzyme activity for different synthetic conditions; iii. stability over time (by measuring size and surface potential variations by DLS) at different temperatures mimicking working (37°C) and storage conditions (-20°C and 4°C).
Overall, data suggest that the produced nanoformulations have optimal physico-chemical characteristics for GALC brain delivery, and good stability in all tested conditions.