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Laronidase-Functionalized Multiple-Wall Lipid-Core Nanocapsules: Promising Formulation for a More Effective Treatment of Mucopolysaccharidosis Type I

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Abstract

Purpose

Mucopolysaccharidosis I is a genetic disorder caused by alpha-L-iduronidase deficiency. Its primary treatment is enzyme replacement therapy (ERT), which has limitations such as a high cost and a need for repeated infusions over the patient's lifetime. Considering that nanotechnological approaches may enhance enzyme delivery to organs and can reduce the dosage thereby enhancing ERT efficiency and/or reducing its cost, we synthesized laronidase surface-functionalized lipid-core nanocapsules (L-MLNC).

Methods

L-MLNCs were synthesized by using a metal complex. Size distributions were evaluated by laser diffraction and dynamic light scattering. The kinetic properties, cytotoxicity, cell uptake mechanisms, clearance profile and biodistribution were evaluated.

Results

Size distributions showed a D[4,3] of 134 nm and a z-average diameter of 71 nm. L-MLNC enhanced the Vmax and Kcat in comparison with laronidase. L-MLNC is not cytotoxic, and nanocapsule uptake by active transport is not only mediated by mannose-6-phosphate receptors. The clearance profile is better for L-MLNC than for laronidase. A biodistribution analysis showed enhanced enzyme activity in different organs within 4 h and 24 h for L-MLNC.

Conclusions

The use of lipid-core nanocapsules as building blocks to synthesize surface-functionalized nanocapsules represents a new platform for producing decorated soft nanoparticles that are able to modify drug biodistribution.

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Abbreviations

BS:

Backscattering

CNS:

Central Nervous System

ERT:

Enzyme Replacement Therapy

IDUA:

Alpha-L-iduronidase

L1-MLNC1 :

Multiple-wall nanocapsule with 0.05% chitosan and 11 μg/mL of laronidase

L1-MLNC2 :

Multiple-wall nanocapsule with 0.075% chitosan and 11 μg/mL of laronidase

L2-MLNC1 :

Multiple-wall nanocapsule with 0.05% chitosan and 96 μg/mL of laronidase

L2-MLNC2 :

Multiple-wall nanocapsule with 0.075% chitosan and 96 μg/mL of laronidase

L-MLNC:

Laronidase surface-functionalized lipid-core nanocapsules

LNC:

Lipid core nanocapsules

LNC-CS0.03 :

LNC coated with 0.03% (w/v) chitosan

LNC-CS0.05 :

LNC coated with 0.05% (w/v) chitosan

LNC-CS0.075 :

LNC coated with 0.075% (w/v) chitosan

MPS I:

Mucopolysaccharidosis type I

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide

PCS:

Photon Correlation Spectroscopy

PDI:

Polydispersity Index

T:

Transmission

∆BS:

Relative Backscattering

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ACKNOWLEDGMENTS AND DISCLOSURES

The authors are grateful for the financial support of the Fundo de Incentivo à Pesquisa (FIPE-HCPA), PRONEX and PRONEM FAPERGS/CNPq, INCT-if CNPq, Universal CNPq, FAPERGS and Rede Nanobiotec CAPES. The sponsors had no involvement in the study design, data collection, analysis, interpretation, writing and decision to publish the data.

Author information

Author notes

  1. Fabiana Quoos Mayer

    Present address: Instituto de Pesquisas Veterinárias Desidério Finamor, Fundação Estadual de Pesquisa Agropecuária, Eldorado do Sul, RS, Brazil

Authors and Affiliations

  1. Gene Therapy Center, Experimental Research Center, Hospital de Clínicas de Porto Alegre, Porto Alegre, RS, Brazil

    Fabiana Quoos Mayer, Talita Giacomet de Carvalho, Anna Cláudia Dilda, Roberto Giugliani & Ursula Matte

  2. Post-Graduation Program on Genetics and Molecular Biology, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

    Fabiana Quoos Mayer, Talita Giacomet de Carvalho, Roberto Giugliani & Ursula Matte

  3. Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

    Márcia Duarte Adorne, Eduardo André Bender, Ruy Carlos Ruver Beck, Sílvia Stanisçuaski Guterres & Adriana Raffin Pohlmann

  4. Medical Genetics Service, HCPA, and Department of Genetics/UFRGS, Porto Alegre, RS, Brazil

    Roberto Giugliani

  5. Departamento de Química Orgânica, Instituto de Química, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

    Adriana Raffin Pohlmann

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  1. Fabiana Quoos Mayer
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Corresponding authors

Correspondence to Ursula Matte or Adriana Raffin Pohlmann.

Additional information

Fabiana Quoos Mayer and Márcia Duarte Adorne contributed equally to this study

Electronic Supplementary Material

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Supplementary figure 1

Particle size distribution profiles (μm) by laser diffraction (Mastersizer 2000, Malvern): (A) LNC-CS0.03; (B) LNC-CS0.05 and (C) LNC-CS0.075. (JPEG 73 kb)

Supplementary figure 2

Relative backscattering profiles of multiple wall nanocapsules over 24 h. A. L1-MLNC1 and B. L2-MLNC2. Only one profile of each suspension is shown. Both formulations showed the same profile. No significant changes were observed in the BS signal at the center of the cuvette. (JPEG 34 kb)

Supplementary figure 3

A cytotoxicity analysis of laronidase, L1-MLNC1 and LNC-CS0.05. Laronidase caused no cytotoxicity at any of the tested concentrations. L1-MLNC1 and LNC-CS0.05 caused approximately 30% cell death at higher concentrations and 90-100% cell viability was reached at concentrations lower than 0.0232 μg/mL. (GIF 55 kb)

High resolution image (TIFF 7605 kb)

Supplementary figure 4

Enzyme uptake by the MPS I patient’s fibroblasts. Laronidase and L1-MLNC1 could be internalized, restoring enzyme activity to the MPS I patient’s fibroblasts. *p < 0.01 compared with Control group, ANOVA with Tukey’s post hoc test. (GIF 21 kb)

High resolution image (TIFF 6883 kb)

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Mayer, F.Q., Adorne, M.D., Bender, E.A. et al. Laronidase-Functionalized Multiple-Wall Lipid-Core Nanocapsules: Promising Formulation for a More Effective Treatment of Mucopolysaccharidosis Type I. Pharm Res 32, 941–954 (2015). https://doi.org/10.1007/s11095-014-1508-y

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  • DOI: https://doi.org/10.1007/s11095-014-1508-y

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