Enzyme-Loaded Gel Core Nanostructured Lipid Carriers to Improve Treatment of Lysosomal Storage Diseases: Formulation and In Vitro Cellular Studies of Elosulfase Alfa-Loaded Systems.

J Álvarez,Andrés Beiras Iglesias,María Couce,Asteria Luzardo Álvarez,Cristóbal Colón Mejeras,Carolina Herrero Filgueira,José Blanco Méndez,Francisco Otero Espinar,Shunji Tomatsu,Alexandre González

doi:10.3390/pharmaceutics11100522

J Álvarez, Andrés Beiras Iglesias + Show 8 more

Open Access

https://doi.org/10.3390/pharmaceutics11100522

Copy DOI

Abstract

Mucopolysaccharidosis IVA (Morquio A) is a rare inherited metabolic disease caused by deficiency of the lysosomal enzyme N-acetylgalatosamine-6-sulfate-sulfatase (GALNS). Until now, treatments employed included hematopoietic stem cell transplantation and enzyme replacement therapy (ERT); the latter being the most commonly used to treat mucopolysaccharidoses, but with serious disadvantages due to rapid degradation and clearance. The purpose of this study was to develop and evaluate the potential of nanostructured lipid carriers (NLCs) by encapsulating elosulfase alfa and preserving its enzyme activity, leading to enhancement of its biological effect in chondrocyte cells. A pegylated elosulfase alfa-loaded NLC was characterized in terms of size, ζ potential, structural lipid composition (DSC and XRD), morphology (TEM microscopy), and stability in human plasma. The final formulation was freeze-dried by selecting the appropriate cryoprotective agent. Viability assays confirmed that NLCs were non-cytotoxic to human fibroblasts. Imaging techniques (confocal and TEM) were used to assess the cellular uptake of NLCs loaded with elosulfase alfa. This study provides evidence that the encapsulated drug exhibits enzyme activity inside the cells. Overall, this study provides a new approach regarding NLCs as a promising delivery system for the encapsulation of elosulfase alfa or other enzymes and the preservation of its activity and stability to be used in enzymatic replacement therapy (ERT).

Highlights

Mucopolysaccharidoses (MPSs) are a group of inherited lysosomal storage disorders (LSDs) associated with deficiencies in lysosomal enzymes and characterized by the accumulation of glycosaminoglycans (GAGs)
Morquio A disease [3,4], is caused by the deficiency of lysosomal enzyme N-acetylgalactosamine 6-sulphatase (GALNS, E.C.3.1.6.4) [5], which leads to a progressive accumulation of the substrate of the enzyme at the cellular level in different tissues, such as bone and cartilage [6,7,8,9,10]
The mixtures of lipids were obtained under similar conditions as reported previously for nanostructured lipid carriers (NLCs) preparation

Summary

Introduction

Mucopolysaccharidoses (MPSs) are a group of inherited lysosomal storage disorders (LSDs) associated with deficiencies in lysosomal enzymes and characterized by the accumulation of glycosaminoglycans (GAGs). MPSs are caused by a deficit of intra-lysosomal specific enzymes or enzymes involved in the transport of proteins from the nucleus to the cytoplasm [1,2]. Morquio A disease (or mucopolysaccharidosis IVA; MPS IVA) [3,4], is caused by the deficiency of lysosomal enzyme N-acetylgalactosamine 6-sulphatase (GALNS, E.C.3.1.6.4) [5], which leads to a progressive accumulation of the substrate of the enzyme at the cellular level in different tissues, such as bone and cartilage [6,7,8,9,10]. The two available therapies for MPS IVA in clinical practice are intravenous administration of the recombinant GALNS enzyme [13,14,15], (elosulfase alfa) to patients weekly (so-called enzyme replacement therapy (ERT)) and hematopoietic stem cell transplantation. ERT with elosulfase alfa is the established treatment for treating somatic symptoms of MPS IVA

Objectives

Methods

Results