Desenvolvimento de nanoparticulas poliméricas carreadoras de proteínas como potenciais adjuvantes para uso em vacinas

Abstract

Nanotechnology has been advancing at the national and international pharmaceutical, seeking the administration of new substances with pharmacological immunological activity. The nanomedicine or the development effective clinics treatments related to nanotechnology is based on controlled release system with purpose of carrying active agents in organisms, tissues or cells, to improve therapeutic efficacy and decrease toxic effects. The nanoparticles are controlled release systems prepared with biodegradable polymers or non-biodegradable, where molecules of therapeutic interest can be dissolved, coated, encapsulated or dispersed in the polymeric matrix. Although many types of polymers present some specific advantages, the lactic and glycolic acids derivates have been studied for use in humans, initially in sutures and orthopedic prostheses and they are commonly used for preparation of nanoparticles mainly by its characteristic of biodegradability and release control. With the aim of dominate the manufacturing process of nanoparticles and to develop an adjuvant for vaccine, was designed and characterized a model for incorporation and encapsulation of proteins. The best formulation was chosen by examining the different methods of obtaining nanoparticles, and equipment used in production, mixing and sonication time, internal aqueous phase volume, polymer concentration, surfactant concentration, protein concentration and cryoprotectant. The parameters investigated for choosing this model and manufacturing process were pH of the preparation, distribution, size, zeta potential of the nanoparticles and encapsulation rate of proteins. The purpose will be to induce immunity, using nanoparticles containing proteins extracted from human epithelial ovarian cancer. The cellular viability in the cell line HEK (Human Kidney embryo) and osteblasts were determined by the colorimetric assay of MTT which consists in the reduction of MTT by mitochondrial dehydrogenase enzyme, present only in viable cells, producing formazan, a blue-violet compound that once precipitated is quantified by spectrophotometry at 595 nm. The results indicate the lack of toxicity of nanoparticles, their components and degradation of substances, however, studies in other cell lines should be conducted. Nanoparticles produced showed low size dispersion, diameter enough to be filtered at 220 nm with high yield, values of zeta potential to provide satisfactory stability of the formulation and encapsulation efficiency of proteins above 70%, indicating a high system capacity. In the present context, pharmaceutical nanotechnology has the potential to be the main instrument in the process of developing new products with advanced technology, solving several problems of therapeutic conventional formulations. The association "of potential therapeutic substance + nanocarrier" represents a paradigm shift for the development of pharmaceuticals, biopharmaceuticals and vaccines, contributing to the increase of scientific production and to generate innovative products.