Targeted Enzymatic VLP‐Nanoreactors with β‐Glucocerebrosidase Activity as Potential Enzyme Replacement Therapy for Gaucher's Disease

Abstract

Gaucher disease is a genetic disorder and the most common lysosomal disease caused by the deficiency of enzyme βglucocerebrosidase (GCase). Although enzyme replacement therapy (ERT) is successfully applied using mannose-exposed conjugated glucocerebrosidase, the lower stability of the enzyme in blood demands periodic intravenous administration that adds to the high cost of treatment. In this work, the enzyme β-glucocerebrosidase was encapsulated inside viruslike nanoparticles (VLPs) from brome mosaic virus (BMV), and their surface was functionalized with mannose groups for targeting to macrophages. The VLP nanoreactors showed significant GCase catalytic activity. Moreover, the Michaelis-Menten constants for the free GCase enzyme (K M = 0.29 mM) and the functionalized nanoreactors (K M = 0.32 mM) were similar even after chemical modification. Importantly, the stability of enzymes under physiological conditions (pH 7.4, 37 °C) was enhanced by � 11-fold after encapsulation; this is beneficial for obtaining a higher blood circulation half-life, which may decrease the cost of therapy by reducing the requirement of multiple intravenous injections. Finally, the mannose receptor targeted enzymatic nanoreactors showed enhanced internalization into macrophage cells. Thus, the catalytic activity and cell targeting suggest the potential of these nanoreactors in ERT of Gaucher's disease.

Gaucher Disease: A Glance from a Medicinal Chemistry Perspective

“…[65] In this perspective, GCase has been successfully encapsulated inside non-infectious virus-like nanoparticles (VLP's) functionalized with mannose groups to be specifically targeted to macrophage cells. [66] The encapsulated GCase was efficiently internalized in the cells, resulting in a significant increase of its activity. Moreover, this enzymatic preparation proved to be more stable compared to the free enzyme, thus resulting in a prolonged half-life in the blood stream which, in turn, can reduce the frequency of the injections and consequently the overall cost of the treatment.…”

“…In this perspective, GCase has been successfully encapsulated inside non‐infectious virus‐like nanoparticles (VLP's) functionalized with mannose groups to be specifically targeted to macrophage cells [66] . The encapsulated GCase was efficiently internalized in the cells, resulting in a significant increase of its activity.…”

Protein‐based Nanoparticles: From Drug Delivery to Imaging, Nanocatalysis and Protein Therapy

“…For additional targeting to macrophage cells, the surface of the 29 nm particles was decorated with mannose‐capped PEG chains. The encapsulation significantly increased the enzyme stability and showed high intracellular GCase activity [22a] …”

Virus-like particles nanoreactors: from catalysis towards bio-applications

“…158,159 As shown in Fig. 5a, to enhance the stability of enzymes in blood demands periodic intravenous administration, Chauhan et al encapsulated the enzyme bglucocerebrosidase (GCase) within VLPs derived from the BMV, 64 for the treatment of Gaucher disease, a common lysosomal disease arising from GCase deficiency. Notably, surface functionalization of the specific receptor is a widely used strategy for precise targeting.…”