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Development of a PEGylated-based Platform for Efficient Delivery of Dietary Antioxidants Across the Blood–Brain Barrier Fernanda M. Borges, Carlos Fernandes, Miguel Pinto, Claudia Martins, Maria João Gomes, Bruno Sarmento, Paulo Oliveira, and Fernando Remião Bioconjugate Chem., Just Accepted Manuscript • DOI: 10.1021/acs.bioconjchem.8b00151 • Publication Date (Web): 11 Apr 2018 Downloaded from http://pubs.acs.org on April 11, 2018
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Bioconjugate Chemistry
Development of a PEGylated-based Platform for Efficient Delivery of Dietary Antioxidants Across the Blood–Brain Barrier Carlos Fernandesa, Miguel Pintoa, Cláudia Martinsb,c, Maria João Gomesb,c,d, Bruno Sarmentob,c,e, Paulo J. Oliveiraf, Fernando Remiãog, Fernanda Borgesa* a
CIQUP, Centro de Investigação em Química, Departmento de Química e Bioquímica,
Faculdade de Ciências, Universidade do Porto, 4169-007, Porto, Portugal. b
i3S, Instituto de Investigação e Inovação em Saúde, Universidade do Porto, 4200-393
Porto, Portugal c
INEB, Instituto de Engenharia Biomédica, Nanomedicines & Translational Drug
Delivery Group, Universidade do Porto, 4200-393 Porto, Portugal d
ICBAS, Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, 4050-
313 Porto, Portugal e
CESPU, Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da
Saúde, 4585-116 Gandra, Portugal f
CNC-Center for Neuroscience and Cell Biology, UC-Biotech, University of Coimbra,
Biocant Park, 3060-197 Cantanhede, Portugal. g
UCIBIO-REQUIMTE, Laboratório de Toxicologia, Departamento de Ciências
Biológicas, Faculdade de Farmácia, Universidade do Porto, 4050-313, Porto, Portugal.
Corresponding Author: Fernanda Borges, CIQUP/Departmento de Química e Bioquímica, Faculdade de Ciências, Universidade do Porto, 4169-007, Porto, Portugal. E-mail:
[email protected] 1 ACS Paragon Plus Environment
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ABSTRACT The uptake and transport of dietary antioxidants remains the most important setback for their application in the therapy. To overcome the limitations a PEGylated-based platform was developed to improve the delivery properties of two dietary hydroxycinnamic (HCA) antioxidants - caffeic and ferulic acids. The antioxidant properties of the new polymerantioxidant conjugates (PEGAntiOxs), prepared by linking polyethylene glycol (PEG) to the cinnamic acids by a one-step Knovenagel condensation reaction, were evaluated. PEGAntiOxs present a higher lipophilicity than the parent compounds (caffeic and ferulic acids) and similar, or higher, antioxidant properties. PEGAntiOxs, were not cytotoxic, at the tested concentrations, in SH-SY5Y, Caco-2 and hCMEC/D3 cells. Contrarily, cytotoxic effects in hCMEC/D3 and SH-SY5Y cells were observed, at 50 and 100 µM, for caffeic and ferulic acids. PEGAntiOxs operate as antioxidants against several oxidative stress-cellular inducers in a neuronal cell-based model, and were able to inhibit glycoprotein-P in Caco-2 cells. PEGAntiOxs can cross hCMEC/D3 monolayer cells, a model of the blood-brain barrier (BBB) endothelial membrane. In summary, PEGAntiOxs are valid antioxidant prototypes that can uphold the antioxidant properties of HCAs, reduce their cytotoxicity and improve their BBB permeability. PEGAntiOxs can be used in a next future as drug candidates to prevent or slow oxidative stress associated to neurodegenerative diseases.
*To whom correspondence should be addressed. Phone: +351 220402560. Fax: +351 220402659. E-mail:
[email protected];
2 ACS Paragon Plus Environment
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Bioconjugate Chemistry
INTRODUCTION Neurodegenerative disorders are amongst the most debilitating diseases of this century and are related with sporadic and/or familial conditions that are characterized by neuronal damage.(1) Although the etiology of these diseases has not yet been fully understood, oxidative stress, induced by imbalanced redox states involving either excessive generation of reactive oxygen species (ROS) or dysfunction of the endogenous antioxidant system, has been linked to the pathophysiological events. Brain is one of the most metabolically active organs vulnerable to oxidative stress, a process that has deleterious effects as it can induce cellular damage, impairment of the DNA repair system and mitochondrial dysfunction, among others. During the progression of age-related neurodegenerative conditions, the capacity of cells to maintain the redox balance decreases and in consequence the cascade of oxidative stress events upsurge contributing for disease progression.(2, 3) As oxidative damage pathways constitute a complex network the blockage at different levels with diverse types of antioxidants can be beneficial. Thus, antioxidant therapy has been proposed as a promising approach for delaying or slowing the neurodegeneration progression.(1) The up-regulation of the pool of endogenous antioxidants or the intake of exogenous dietary antioxidants, such as hydroxycinnamic acids (HCAs), have been stated as an effective strategy for delaying or slowing the degenerative progression of neurons.(4) In fact, it is assumed that a regular consumption of fruits and vegetables, commonly attributed to the phenolic antioxidants, is linked with neuroprotective benefits. However, the majority of antioxidants studied so far have limited success in clinical trials, a fact that could be related with their poor distribution and with the inherent difficulties to cross the brain-blood barrier (BBB) and attain the target sites.(5) The BBB is considered the most important barrier to protect the brain, constituting a rate-limiting factor in the transport of drugs into the central nervous system.(6) The large BBB surface area of the endothelial membrane offers an effective transcellular passive diffusion pathway for lipid-soluble agents and small gaseous molecules, whereas the entry of hydrophilic molecules is generally repressed.(7) Around 98 % of small molecules (molecular weight
