Novel lipid nanovesicle-loaded dissolving microarray patches for fenretinide in breast cancer chemoprevention.

The retinoid fenretinide (FENR) is a promising compound for preventing breast cancer recurrence but faces challenges due to poor solubility and low bioavailability. This study explores the development of dissolving microneedles (MNs) containing FENR-loaded ethosomes for minimally invasive breast cancer chemoprevention, aiming to enhance local drug distribution. Ethosomes were formulated using ethanol, propylene glycol, soya lecithin, water, and polysorbate 80 micelles. MNs were created from poly(vinyl alcohol) and poly(vinylpyrrolidone) hydrogels by adding polymer powder directly into ethosomes suspensions, reducing manufacturing time and cost. Two methods were used to load ethosomes into high-density moulds: 1) only in the needle area, and 2) in both the needle area and baseplate. Dynamic light scattering confirmed nanostructures in the hydrogels and MNs. Micelle-based ethosomes dissolved MNs in 15 min, compared to 30 min for other MNs. Skin deposition studies showed greater drug deposition (up to 10 µg/patch) and enhanced skin permeation of FENR (up to 40 µg) with Method 2. In-vivo studies in rats demonstrated that oral administration resulted in plasma FENR levels below 10 ng/g in the first three hours, whereas MN administration delayed delivery, reaching a maximum plasma concentration of 52 ng/g at 48 h. Skin deposition of FENR from MNs decreased from 3 µg/g on day 1 to <0.3 µg/g by the last day. This study indicates that MNs are a potential minimally invasive dosage form for delivering FENR, offering a new approach for breast cancer chemoprevention.

Rethinking Breast Cancer Chemoprevention: Technological Advantages and Enhanced Performance of a Nanoethosomal-Based Hydrogel for Topical Administration of Fenretinide.

Herein, we developed an ethosomal hydrogel based on three types of ethosomes: simple, mixed (surfactant-based micelles and lipid vesicles) or binary (comprising two type of alcohols). Ethanol injection was employed for vesicles preparation, and sodium alginate, as gelling agent. We purposed the local-transdermal administration of the off-the-shelf retinoid fenretinide (FENR) for chemoprevention of breast cancer. Rheograms and flow index values for alginate dispersion (without ethosomes) and hydrogels containing simple, mixed or binary ethosomes suggested pseudoplastic behavior. An increase in the apparent viscosity was observed upon ethosome incorporation. The ethosomal hydrogel displayed increased bioadhesion compared to the alginate dispersion, suggesting that the lipid vesicles contribute to the gelling and bioadhesion processes. In the Hen's Egg Test-Chorioallantoic Membrane model, few spots of lysis and hemorrhage were observed for formulations containing simple (score of 2) and mixed vesicles (score 4), but not for the hydrogel based on the binary system, indicating its lower irritation potential. The binary ethosomal hydrogel provided a slower FENR in vitro release and delivered 2.6-fold less drug into viable skin layers compared to the ethosome dispersion, supporting the ability of the gel matrix to slow down drug release. The ethosomal hydrogel decreased by ~ five-fold the IC50 values of FENR in MCF-7 cells. In conclusion, binary ethosomal gels presented technological advantages, provided sustained drug release and skin penetration, and did not preclude drug cytotoxic effects, supporting their potential applicability as topical chemopreventive systems.

Quality by design enabled the development of stable and effective oil-in-water emulsions at compounding pharmacy: the case of a sunscreen formulation.

It is widely accepted that the use of topical sunscreens has medical importance with potential to prevent skin damage by protecting from solar ultraviolet radiation (UVR) effects. Pharmaceutical emulsions require an optimal qualitative and quantitative combination of emollients, emulsifiers and others compounds such as softening agents and, for sunscreens, a combination of chemical and physical UV filters. Herein, we applied the quality by design (QbD) concept to achieve stable and effective compounded sunscreen emulsions. By using the statistical tool of design of experiments, it was possible to identify the influence of emulsifier type (with low and high Hydrophile-Lipophile Balance) and concentrations of emollient and softening agent on the achievement of formulations with suitable organoleptic and physicochemical features. Compounded emulsions with pleasant macroscopic aspects were obtained. Three formulations with physicochemical properties in targeted ranges were selected, namely pH ∼6.0, conductivity > 0.0 µS/cm2, spreadability factor ∼1-1.5 g/mm2, viscosity ∼12000 mPa.s and sunscreen protection factor ∼30. Freeze-thaw cycle and accelerated stability study under different storage conditions allowed selecting a stable emulsion that ensured photoprotection in biological assays. The QbD approach was essential to select the best, low-cost compounded sunscreen emulsion, with targeted physicochemical parameters.

Topical transdermal chemoprevention of breast cancer: where will nanomedical approaches deliver us?

Despite the high incidence of breast cancer, there are few pharmacological prevention strategies for the high-risk population and those that are available have low adherence. Strategies that deliver drugs directly to the breasts may increase drug local concentrations, improving efficacy, safety and acceptance. The skin of the breast has been proposed as an administration route for local transdermal therapy, which may improve drug levels in the mammary tissue, due to both deep local penetration and percutaneous absorption. In this review, we discuss the application of nanotechnology-based strategies for the delivery of well established and new agents as well as drug repurposing using the topical transdermal route to improve the outcomes of preventive therapy for breast cancer.

Lipid nanovesicles for biomedical applications: 'What is in a name'?

Vesicles, generally defined as self-assembled structures formed by single or multiple concentric bilayers that surround an aqueous core, have been widely used for biomedical applications. They can either occur naturally (e.g. exosomes) or be produced artificially and range from the micrometric scale to the nanoscale. One the most well-known vesicle is the liposome, largely employed as a drug delivery nanocarrier. Liposomes have been modified along the years to improve physicochemical and biological features, resulting in long-circulating, ligand-targeted and stimuli-responsive liposomes, among others. In this process, new nomenclatures were reported in an extensive literature. In many instances, the new names suggest the emergence of a new nanocarrier, which have caused confusion as to whether the vesicles are indeed new entities or could simply be considered modified liposomes. Herein, we discussed the extensive nomenclature of vesicles based on the suffix "some" that are employed for drug delivery and composed of various types and proportions of lipids and others amphiphilic compounds. New names have most often been selected based on changes of vesicle lipid composition, but the payload, structural complexity (e.g. multicompartment) and new/improved proprieties (e.g. elasticity) have also inspired new vesicle names. Based on this discussion, we suggested a rational classification for vesicles.

Quality-by-Design Approach for Biological API Encapsulation into Polymersomes Using "Off-the-Shelf" Materials: a Study on L-Asparaginase.

Polymersomes are versatile nanostructures for protein delivery with hydrophilic core suitable for large biomolecule encapsulation and protective stable corona. Nonetheless, pharmaceutical products based on polymersomes are not available in the market, yet. Here, using commercially available copolymers, we investigated the encapsulation of the active pharmaceutical ingredient (API) L-asparaginase, an enzyme used to treat acute lymphoblastic leukemia, in polymersomes through a quality-by-design (QbD) approach. This allows for streamlining of processes required for improved bioavailability and pharmaceutical activity. Polymersomes were prepared by bottom-up (temperature switch) and top-down (film hydration) methods employing the diblock copolymers poly(ethylene oxide)-poly(lactic acid) (PEG45-PLA69, PEG114-PLA153, and PEG114-PLA180) and the triblock Pluronic® L-121 (poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide), PEG5-PPO68-PEG5). Quality Target Product Profile (QTPP), Critical Quality Attributes (CQAs), Critical Process Parameters (CPPs), and the risk assessment were discussed for the early phase of polymersome development. An Ishikawa diagram was elaborated focusing on analytical methods, raw materials, and processes for polymersome preparation and L-asparaginase encapsulation. PEG-PLA resulted in diluted polymersomes systems. Nonetheless, a much higher yield of Pluronic® L-121 polymersomes of 200 nm were produced by temperature switch, reaching 5% encapsulation efficiency. Based on these results, a risk estimation matrix was created for an initial risk assessment, which can help in the future development of other polymersome systems with biological APIs nanoencapsulated.

Poly (lactic-co-glycolic acid) nanospheres allow for high l-asparaginase encapsulation yield and activity.

l-Asparaginase (ASNase) is an amidohydrolase used as a chemotherapeutic agent for the treatment of acute lymphoblastic leukemia (ALL). The nanoencapsulation of this enzyme is strategic to avoid its immediate immunogenic effects that lead to a decrease in the enzyme half-life. In this work, ASNase-containing nanoparticles (NPs) were prepared by double emulsification, through an ultrasonic sonicator or an Ultra-Turrax, using two copolymers of 50:50 (w/w) poly (lactic-co-glycolic acid) (PLGA) with different ranges of molecular weight (24-38 kDa and 30-60 kDa) and varying the concentration of polyvinyl alcohol (PVA) as a stabilizer (0.5, 1.0, 1.5 and 2.0%) as well as the emulsification time (30 and 60 s). Using 24-38 kDa PLGA and 1.0% PVA, we obtained by cavitation NPs with hydrodynamic diameter of 384 nm, polydispersity index of 0.143 and Zeta potential of -16.4 mV, whose ASNase encapsulation efficiency was as high as 87 ±â€¯2%. The encapsulated enzyme showed an activity 22% higher than that of the free enzyme, and no conformational changes were detected by circular dichroism. The enzyme release from NPs entrapped in dialysis bags (500 kDa molecular weight cut-off) allowed selecting a controlled system able to release about 60% of the enzyme within 14 days, for which the Korsmeyer-Peppas model provided the best correlation (R2 = 0.966).

Challenges for the Self-Assembly of Poly(Ethylene Glycol)⁻Poly(Lactic Acid) (PEG-PLA) into Polymersomes: Beyond the Theoretical Paradigms.

Polymersomes (PL), vesicles formed by self-assembly of amphiphilic block copolymers, have been described as promising nanosystems for drug delivery, especially of biomolecules. The film hydration method (FH) is widely used for PL preparation, however, it often requires long hydration times and commonly results in broad size distribution. In this work, we describe the challenges of the self-assembly of poly (ethylene glycol)-poly(lactic acid) (PEG-PLA) into PL by FH exploring different hydrophilic volume fraction (f) values of this copolymer, stirring times, temperatures and post-FH steps in an attempt to reduce broad size distribution of the nanostructures. We demonstrate that, alongside f value, the methods employed for hydration and post-film steps influence the PEG-PLA self-assembly into PL. With initial FH, we found high PDI values (>0.4). However, post-hydration centrifugation significantly reduced PDI to 0.280. Moreover, extrusion at higher concentrations resulted in further improvement of the monodispersity of the samples and narrow size distribution. For PL prepared at concentration of 0.1% (m/v), extrusion resulted in the narrower size distributions corresponding to PDI values of 0.345, 0.144 and 0.081 for PEG45-PLA69, PEG114-PLA153 and PEG114-PLA180, respectively. Additionally, we demonstrated that copolymers with smaller f resulted in larger PL and, therefore, higher encapsulation efficiency (EE%) for proteins, since larger vesicles enclose larger aqueous volumes.

Development of L-Asparaginase Biobetters: Current Research Status and Review of the Desirable Quality Profiles.

L-Asparaginase (ASNase) is a vital component of the first line treatment of acute lymphoblastic leukemia (ALL), an aggressive type of blood cancer expected to afflict over 53,000 people worldwide by 2020. More recently, ASNase has also been shown to have potential for preventing metastasis from solid tumors. The ASNase treatment is, however, characterized by a plethora of potential side effects, ranging from immune reactions to severe toxicity. Consequently, in accordance with Quality-by-Design (QbD) principles, ingenious new products tailored to minimize adverse reactions while increasing patient survival have been devised. In the following pages, the reader is invited for a brief discussion on the most recent developments in this field. Firstly, the review presents an outline of the recent improvements on the manufacturing and formulation processes, which can severely influence important aspects of the product quality profile, such as contamination, aggregation and enzymatic activity. Following, the most recent advances in protein engineering applied to the development of biobetter ASNases (i.e., with reduced glutaminase activity, proteolysis resistant and less immunogenic) using techniques such as site-directed mutagenesis, molecular dynamics, PEGylation, PASylation and bioconjugation are discussed. Afterwards, the attention is shifted toward nanomedicine including technologies such as encapsulation and immobilization, which aim at improving ASNase pharmacokinetics. Besides discussing the results of the most innovative and representative academic research, the review provides an overview of the products already available on the market or in the latest stages of development. With this, the review is intended to provide a solid background for the current product development and underpin the discussions on the target quality profile of future ASNase-based pharmaceuticals.

Desenvolvimento e caracterização de polimerossomos para veiculação de L-asparaginase / Development and characterization of polymersomes for the release of L-asparaginase

A enzima L-Asparaginase (ASNase) é um biofámaco utilizado no tratamento da leucemia linfoblástica aguda, no entanto, a evolução da produção da ASNase como um medicamento desde o final da década de 1970 resultou em apenas quatro alternativas disponíveis no mercado farmacêutico, com relatos de graves reações imunogênicas e toxicidade. Desse modo, a nanotecnologia é uma plataforma que pode ser explorada para administração dessa enzima diminuindo a exposição da mesma a proteases e aumentando a sua meia-vida aparente. Os polimerossomos (PL) são opções que pela nanoestrutura vesicular poderiam encapsular a ASNase em seu core aquoso e pela presença de uma membrana polimérica, são mais robustos que os lipossomos. Assim, neste trabalho objetivou-se desenvolver PL para encapsulação da ASNase como uma alternativa às formulações deste biofármaco existentes. Foram desenvolvidos PL de PEG-PLA, PMPC-PDPA, PEG-PDPA e Pluronic® L-21. Foram estudados fatores relacionados à composição dos copolímeros (fração hidrofílica, responsividade a fatores externos tais como pH e temperatura) e métodos de elaboração (hidratação do filme polimérico, troca de pH e temperatura) bem como foi feita a caracterização dos PL obtidos (tamanho, índice de polidispersão, espessura de membrana, formação de excessivo bulk polimérico, obtenção de micelas). Também foi feito um planejamento racional para encapsulação da ASNase (hidratação direta do filme polimérico e encapsulação por eletroporação, autoagregação com encapsulação por troca de pH ou de temperatura). Para os PL preparados com PEG-PLA, a extrusão resultou em distribuição de tamanhos mais estreitos correspondentes aos valores de PDI de 0,345, 0,144 e 0,081 para PEG45-PLA69, PEG114-PLA153 e PEG114-PLA180, respectivamente. Foi demonstrado que copolímeros com menor fração hidrofóbica resultam em maior eficiência de encapsulação para proteínas, já que possuem volumes aquosos maiores. Com o PMPC25-PDPA72 foi possível encapsular em média três unidades de ASNase por vesículas através da eletroporação ou troca de pH, sendo que no primeiro método houve formação de túbulos e no último método as micelas não foram completamente removidas. Para PEG100-PDPA80, grandes agregados permaneceram após a purificação levando a um PDI alto, mas não foi observada a formação de túbulos, já a troca de pH para este copolímero resultou em maior perda de copolímeros como bulk polimérico precipitado. Para o copolimero tribloco Pluronic® L-121, foi observado que as vesículas eram estáveis durante uma semana à temperatura ambiente, contrariando o que era descrito na literatura. Nesses sistemas, quando preparados por hidratação do filme, a encapsulação da ASNase foi realizada por eletroporação mas a proteína não foi detectada dentro das vesículas. Atribuímos a não-encapsulação à organização da bicamada Pluronic® L-121 sem conformação definida das cadeias poliméricas, dificultando a reorganização do bloco hidrofílico na porção interna do poro durante eletroporação. Por troca de temperatura, cerca de 5 % de ASNase foi encapsulada e o método resultou em total recuperação da atividade da enzima. Desse modo foram obtidos diferentes PL com diferentes características nanoestruturais de acordo com os copolímeros utilizados para carreamento da ASNase

The enzyme L-Asparaginase (ASNase) is a biopharmaceutical used in the treatment of acute lymphoblastic leukemia, still the industrial production of ASNase as a marketable drug since the late 1970s has resulted in only four alternatives available in the pharmaceutical market, with reports of severe immunogenic reactions and toxicity. In this sense, nanotechnology is a platform that can be exploited to administer this enzyme by decreasing its exposure to proteases and increasing its apparent half-life. Polymerosomes (PL) are interesting routes which by its intrinsically vesicular nanostructure could encapsulate the ASNase in its aqueous core and by the presence of a polymeric membrane, being more robust than the liposomes. Thus, in this work it was intended to develop PL for ASNase encapsulation as an alternative to existing formulations of this biopharmaceutical. PL of PEG-PLA, PMPC-PDPA, PEG-PDPA and Pluronic® L-21 were developed. It was studied the copolymers composition (i.e. hydrophilic fraction, responsiveness to external factors such as pH and temperature), PL design (i.e. polymer film hydration, pH change and temperature) and PL characterization (i.e. size, polydispersity index - PDI, membrane thickness, formation of excessive polymer bulk, micelles production). A suitable experimental planning for ASNase encapsulation (i.e. direct hydration of the polymeric film and encapsulation by electroporation, self-aggregation with encapsulation by pH or temperature change) was also performed. For the PL prepared with PEG-PLA, the extrusion resulted in narrower size distribution corresponding to the PDI values of 0.345, 0.144 and 0.081 for PEG45-PLA69, PEG114-PLA153 and PEG114-PLA180, respectively. It has been shown that copolymers with lower hydrophobic fraction result in higher encapsulation efficiency for proteins, since they have larger aqueous volumes. With PMPC25-PDPA72 PL, it was possible to encapsulate three units of ASNase per vesicles through electroporation or pH change. In the first method, tubules were formed and in the latter one the micelles were not completely removed. For PEO100-PDPA80 PL, large aggregates remained after purification leading to a high PDI value, nevertheless no tubule formation was observed, since the pH change for this copolymer resulted in greater loss of copolymers as a precipitated polymer bulk. For the Pluronic® L-121 triblock copolymer PL, it was observed that the vesicles were stable for one week at room temperature, contrary to what was described in the literature. These PLs were prepared by film hydration method and ASNase encapsulation was performed by electroporation, nonetheless the protein was not detected within the vesicles. It is attributed the non-encapsulation to the organization of the Pluronic® L-121 bilayer without defined conformation of the polymer chains, making it difficult to reorganize the hydrophilic block in the internal portion of the pore during electroporation. By temperature change, about 5% of ASNase was encapsulated and the method resulted in complete recovery of enzyme activity. In conclusion, several PLs with a vast range of differential nanostructural characteristics were obtained according to the copolymers used for ASNase loading

Nanostructures for protein drug delivery.

Use of nanoscale devices as carriers for drugs and imaging agents has been extensively investigated and successful examples can already be found in therapy. In parallel, recombinant DNA technology together with molecular biology has opened up numerous possibilities for the large-scale production of many proteins of pharmaceutical interest, reflecting in the exponentially growing number of drugs of biotechnological origin. When we consider protein drugs, however, there are specific criteria to take into account to select adequate nanostructured systems as drug carriers. In this review, we highlight the main features, advantages, drawbacks and recent developments of nanostructures for protein encapsulation, such as nanoemulsions, liposomes, polymersomes, single-protein nanocapsules and hydrogel nanoparticles. We also discuss the importance of nanoparticle stabilization, as well as future opportunities and challenges in nanostructures for protein drug delivery.

Physico-chemical quality parameters of herbal products from Agave sisalana.

Agave sisalana components have great potential in different pharmaceutical applications, but the quality of herbal raw materials is essential to reach the desired product specifications. In this work, we investigated the physico-chemical quality parameters of bole and wastes from decortication of A. sisalana leaves. The statistically significant variations among products suggest different pharmaceutical applications for each of them.

Inulin-type fructans: a review on different aspects of biochemical and pharmaceutical technology.

Inulin is a natural storage polysaccharide with a large variety of food and pharmaceutical applications. It is widely distributed in plants, being present as storage carbohydrate in more than 30,000 vegetable products. Due to their wide distribution in nature and significant role in industry, the extraction, isolation and characterization of inulin-type fructans are gaining attention in recent years. Inulin sources have recently received increasing interest as they are a renewable raw material for the production of bioethanol, fructose syrup, single-cell protein and single cell oil, obtainment of fructooligosaccharides and other useful products. This review focuses on the state-of-the-art of biochemical and pharmaceutical technology of inulin-type fructans.

Uso de cosméticos por mulheres do município de Esperança, Paraíba, Brasil / Use of cosmetics for women of the city of Hope, Paraíba, Brazil

O aumento de vendas de produtos cosméticos e de higiene pessoal deve ser atentamente acompanhado pelos setores regulatórios competentes, uma vez que mais de oito mil matérias-primas são usadas na formulação destes produtos e muitas podem ocasionar efeitos adversos tóxicos ou mesmo alergias. Assim, este trabalho visou investigar quais as finalidades do uso de cosméticos entre mulheres, conhecer os mais utilizados e saber os locais de aquisição destes produtos. Tratou-se de um estudo transversal, descritivo e exploratório, tendo como instrumento de coleta de dados um questionário com perguntas objetivas, aplicado entre 102 mulheres. A higiene (65,7%) foi respondida como a principal finalidade de uso. Os produtos com maior frequência de utilização foram os perfumes e desodorantes (90,2%), maquiagens (74,5%), produtos para depilação (74,5%) e tinturas de cabelos (41,2%). A maior parte das mulheres (63,7%) relatou comprar os cosméticos em revistas. Diante dos resultados expostos, pode-se inferir que as mulheres utilizam variados tipos de cosméticos, com diferentes finalidades, o que pode implicar em risco para saúde.

The increasing sales of cosmetics and toiletries must be carefully monitored by competent regulatory sectors, as many of the more than eight thousand ingredients used to formulate cosmetics can have adverse effects, such as carcinogenicity, genotoxicity, phototoxicity, contact dermatitis, hives, hypopigmentation, hyperpigmentation, depigmentation, damage to hair and nails, etc. The aims of the present study were to investigate the reasons for the use of cosmetics by women and determine both the most frequently used products and where such products are acquired. For such, a cross-sectional study with an analytical approach was carried out. Data were collected through the use of a questionnaire with objective questions administered to 102 women. Hygiene (65.7%) was the main reason cited for using cosmetic products. The most frequently used products were perfumes and deodorants (90.2%), makeup (74.5%), hair removal products (74.5%) and hair dyes (41.2%). The majority (63.7%) reported to buying cosmetics from catalogues. The present findings demonstrate that women use many kinds of cosmetics for different purposes, which may imply health risks.