Impact of Fluorinated Ionic Liquids on Human Phenylalanine Hydroxylase—A Potential Drug Delivery System

Abstract

Phenylketonuria (PKU) is an autosomal recessive disease caused by deficient activity of human phenylalanine hydroxylase (hPAH), which can lead to neurologic impairments in untreated patients. Although some therapies are already available for PKU, these are not without drawbacks. Enzyme-replacement therapy through the delivery of functional hPAH could be a promising strategy. In this work, biophysical methods were used to evaluate the potential of [N1112(OH)][C4F9SO3], a biocompatible fluorinated ionic liquid (FIL), as a delivery system of hPAH. The results herein presented show that [N1112(OH)][C4F9SO3] spontaneously forms micelles in a solution that can encapsulate hPAH. This FIL has no significant effect on the secondary structure of hPAH and is able to increase its enzymatic activity, despite the negative impact on protein thermostability. The influence of [N1112(OH)][C4F9SO3] on the complex oligomerization equilibrium of hPAH was also assessed.

Simultaneous Purification of Human Interferon Alpha-2b and Serum Albumin Using Bioprivileged Fluorinated Ionic Liquid-Based Aqueous Biphasic Systems

“…The development of functionalized, bioprivileged, task-specific FILs was previously achieved through the increasing of the perfluoroalkyl sulfonate anion chain ([C F SO ] or greater) [ 37 , 67 , 71 , 72 , 73 , 74 , 75 , 76 , 77 ]. These FILs have demonstrated distinct transitions associated with the formation of stable self-assembled structures, which might explain its complete miscibility in water [ 71 ].…”

“…The FILs, based on the [C F SO ] anion implemented in this study, demonstrated to be biocompatible and non-toxic in four human cell lines (Caco-2, HepG2, EA.hy926, and HaCaT) and aquatic species with different biological organization levels ( Vibrio fischeri , Daphnia magna , and Lemna minor ) [ 72 , 73 ]. The effects of these biocompatible FILs using lysozyme, BSA, human phenylalanine hydroxylase, and IFN- 2b as model proteins [ 37 , 74 , 75 , 76 , 77 ] were evaluated, proving the [C F SO ]-based FILs as promising potential biomaterials for drug delivery systems using therapeutic proteins. The formation of aqueous biphasic systems using these biocompatible FILs, based on perfluoroalkyl sulfonate anions, was first proposed by one of the authors [ 54 ] as an extractive platform, proving its potential for the efficient and selective extraction of food colorants.…”

“…The efficacy and safety of therapeutic proteins are related to the folded conformation of the protein, and different manipulations in extraction or purification steps have the potential to alter the active ingredient. DSC is used in different studies to address protein folding and stability [ 99 ], as well as CD, an analytical method often used to examine structural changes that occur when a protein interacts with another molecule [ 37 , 74 , 76 , 77 , 100 ] (and references cited therein). Herein, the impact of the ABS phase on the stability and structure of IFN- 2b and BSA was assessed and compared with the behavior of the proteins in water.…”

“…Additionally, they were shown to be biocompatible and non-toxic in four human cell lines (Caco-2, HepG2, EA.hy926, and HaCaT) and aquatic species with different levels of biological organization (Vibrio fischeri, Daphnia magna, and Lemna minor) [ 72 , 73 ]. Further, the effect of these biocompatible FILs using lysozyme, BSA, human phenylalanine hydroxylase, and IFN- 2b as model proteins [ 37 , 74 , 75 , 76 , 77 ] was evaluated, showcasing these FILs as promising potential biomaterials for drug delivery systems using therapeutic proteins. Finally, the functionalization of ABS was attained through the development of biocompatible ABS based on these perfluoroalkyl sulfonate FILs for the selective partition of lysozyme as the model protein [ 78 ].…”

Disclosing the Potential of Fluorinated Ionic Liquids as Interferon-Alpha 2b Delivery Systems

“…The results showed that the self-assembled structures of FILs have strong interactions with BSA, and they can be considered a promising potential novel biomaterial for drug delivery systems using therapeutic proteins [ 43 ]. Recently, the study of the interactions between FILs and human phenylalanine hydroxylase protein has also demonstrated the encapsulation of the protein by the FIL aggregates [ 44 ]. Therefore, the encapsulation and stabilization of therapeutic proteins using FILs have demonstrated to be a promising strategy to overcome the handicaps in the delivery of therapeutic proteins in the human body, allowing the improvement of their bioavailability, safety, and efficacy by preserving the structure, stability, and biological activity of several different biomolecules.…”