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Since psoriasis is an immuno-mediated skin disease, long-term therapies are necessary for its treatment. In clinical investigations, tacrolimus (TAC), a macrolide immunosuppressive inhibitor of calcineurin, arises as an alternative for the treatment of psoriasis, acting in some cytokines involved in the pathogenesis of the disease. Here, we aim to study the psoriasis treatment with TAC and siRNA for one of most cytokines expressed in psoriasis, the TNF-alpha. A multifunctional nanostructure lipid carrier (NLC) was developed to co-delivery TAC and siRNA. Results showed that the particle size and zeta potential were around 230 nm and + 10 mV, respectively. The release study demonstrated a controlled release of TAC, and the permeation and retention profile in the skin tissue show to be promising for topical application. The cell viability and uptake in murine fibroblast presented low toxicity associated to uptake of NLC in 4 h, and finally, the in vivo animal model demonstrates the efficiency of the NLC multifunctional, exhibiting a reduction of the cytokine TNF-alpha expression about 7-fold and presenting a synergic effect between the TAC and TNF-alpha siRNA. The developed system was successfully to treat in vivo psoriatic animal model induced by imiquimod and the synergic combination was reported here for the first time.Graphical abstract

Celecoxib (CXB) is a widely used anti-inflammatory drug that also acts as a chemopreventive agent against several types of cancer, including skin cancer. As the long-term oral administration of CXB has been associated with severe side effects, the skin delivery of this drug represents a promising alternative for the treatment of skin inflammatory conditions and chemoprevention of skin cancer. We prepared and characterized liquid crystalline systems based on glyceryl monooleate and water containing penetration enhancers which were primarily designed to promote skin delivery of CXB. Analysis of their phase behavior revealed the formation of cubic and hexagonal phases depending on the systems' composition. The systems' structure and composition markedly affected the in vitro CXB release profile. Oleic acid reduced CXB release rate, but association oleic acid/propylene glycol increased the drug release rate. The developed systems significantly reduced inflammation in an aerosil-induced rat paw edema model. The systems' composition and liquid crystalline structure influenced their anti-inflammatory potency. Cubic phase systems containing oleic acid/propylene glycol association reduced edema in a sustained manner, indicating that they modulate CXB release and permeation. Our findings demonstrate that the developed liquid crystalline systems are potential carriers for the skin delivery of CXB.

In this study, we developed a water-in-oil microemulsion containing vitamin A (retinol) and vitamin E (α-tocopherol), which serves as a multifunctional nanosystem that co-delivers antioxidants and displayed additive effect against acute skin inflammation. Microemulsion (ME) was prepared by mixing a surfactant blend (Tween 80 and propylene glycol, 5:1) with isopropyl myristate and water (ratio of 50:40:10, respectively). Vitamin A (0.05% w/w concentration) and/or vitamin E (0.1% w/w concentration) were incorporated into the surfactant mixture of ME by stirring with a magnetic stirrer for 30 min. This multifunctional ME displayed physical stability, with low cytotoxicity in 3T3 cell line, as well as cellular internalization into the cytosol. In vivo treatments using ME delivering α-tocopherol reduced dermal expression of TNF-α by 1.3-fold (p 0.01), when compared to unloaded ME treatment group. When retinol was added into the ME containing α-tocopherol, it further reduced TNF-α expression by 2-fold (p 0.001), suggesting the additive effect of vitamin E and vitamin A in the treatment against skin inflammation. In conclusion, we successfully developed the use of water-in-oil ME to pack both vitamin E and vitamin A, and demonstrated for the first time its anti-inflammatory potential when applied topically to TPA-induced inflamed skin.

Introduction: There are no published studies on adverse drug reactions (ADRs) as a cause of inpatient treatment for Brazilian patients. Although the Dose-Time Sensitivity Classification System has been shown to be useful for assessing ADRs in hospitalized patients. Objective: We analyzed the cases of hospitalization due to drug poisoning attending the hospitals. Methods: Standardized cases reports of 47 patients undergoing clinical therapy in hospital Padre Albino and Emilio Carlos from 2010 to 2015 were used as data collection tool to support this investigation. Results: A total 47 patients were admitted to the hospitals for exogenous intoxication, 81%, of these patients, had at least one ADR as the cause of hospital admission, with a total of a single drug reaction. The mean age of patients who experienced ADRs was about 42.31 years, among patients with ADRs, 39% were female, (47.13years), and male (39.17years). None of the women were pregnant. Conclusions: We found that the patients took between 1 and 6 medicatins and the reason for hospitalization of these patients in the countryside of the Brazil was the ADRs incidence. Our results is in the agreement with previously reported by other coutries.

Made available in DSpace on 2018-12-11T17:15:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-01-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Ministry of Science and Technology In vitro skin permeation/penetration studies may be affected by many sources of variation. Herein, we aimed to investigate the major critical procedures of in vitro skin delivery studies. These experiments were performed with model drugs according to official guidelines. The influence of skin source on penetration studies was studied as well as the use of a cryopreservation agent on skin freezing evaluated by transepidermal water loss, electrical resistance, permeation/penetration profiles and histological changes of the skin. The best condition for tape stripping procedure was validated through the evaluation of the distribution of corneocytes, mass of stratum corneum (SC) removed and amount of protein removed using finger pressure, a 2 kg weight and a roller. The interchangeability of the tape stripping procedures followed by the epidermis and dermis homogenate and the micrometric horizontal cryostat skin sectioning methods were also investigated, besides the effect of different formulations. Noteworthy, different skin sources were able to ensure reliable interchangeability for in vitro permeation studies. Furthermore, an increased penetration was obtained for stored frozen skin compared to fresh skin, even with the addition of a cryoprotectant agent. The best method for tape stripping was the finger pressure followed by the addition of a propylene glycol solvent leading to better SC removal. Finally, no significant difference was found in skin penetration studies performed by different methods suggesting their possible interchangeability. School of Pharmaceutical Sciences of Ribeirão Preto University of São Paulo, Avenida do Café, s/n Faculdades Integradas Padre Albino – FIPA Research Institute for Medicines (iMed.ULisboa) Faculdade de Farmácia University of Lisbon School of Pharmaceutical Sciences UNESP – São Paulo State University Campus Araraquara Department of Drugs and Medicines, Rodovia Araraquara-Jau, km. 1 School of Pharmaceutical Sciences UNESP – São Paulo State University Campus Araraquara Department of Drugs and Medicines, Rodovia Araraquara-Jau, km. 1 FAPESP: 04/09465-7 FAPESP: 2013/16617-1

Introduction: Diagnosis of immediate hypersensitivity reaction (HSR) for both iodinated contrast (ICM) and non ionic contrast media(NICM) is challenging based on clinical history and skin tests. Objective: The goal of this study was evaluate the negative predictive value for patients with suspected HSR to ICM and NICM doses. Methods: Standardized cases reports and clinical history of hundred and forty-one patients undergoing clinical therapy in Department of Radiology at Padre Albino Hospital from December 2015 to April 2016 were used as data collection tool to support this investigation. Results: Fourty-five from 141 patients investigated were challenged by ICM and only one patient had a positive challenge result (2.2%)which was a grade I reaction by the Ring and Messmer classification. Thirty from 45 patients evaluated were re-exposed to ICM during radiologic examination (28%) and none of them experienced a negativechallenge to ICM when a premedication or ICM replacement by NICM was applied. On the other hand, ninety six from 141 patients investigated were selected to challenge successfully NICM. Conclusions: In our study few patients had documented clinical history of immediate response to ICM and they received a premedication before re-exposure to ICM or even received ICM replacement by NICM. In conclusion, our results showed that clinical decision based on patient clinical records resulted in a decrease of several immediate reactions incidence

Topical photodynamic therapy (PDT) involves both local administration of a photosensitizing drug followed by illumination or irradiation of visible light, resulting in light-drug combinations to kill cells and microorganisms. PDT using tetra-sulfonate phthalocyanine, a second generation of hydrophilic photosensitizers, has recently become a good treatment option for treatment of skin malignances. However, tetra-sulfonate phthalocyanine must be delivered into the target layers of the skin to achieve promising results. Thus, strategies to improve the phthalocyanine cell uptake based on lipid-based nanocarriers were evaluated in this work. The results showed that tetra-sulfonate phthalocyanine loaded lipid-based nanocarriers (microemulsion and liquid crystalline nanodispersion) have nanometric particle size, with low polydispersity, negative zeta potential and high ZnPcSO4 encapsulation efficiency. Furthermore, both nanocarriers investigated presented little cytotoxicity against HaCaT cells, however the microemulsion presented higher toxicity upon higher concentrations. Finally, the formulations developed herein enabled high ZnPcSO4 uptake, a promising result. Future studies will be conducted using tumor skin cells lines.

Breast cancer is the most frequently diagnosed cancer and the leading cause of cancer death in women worldwide. Chemotherapy against breast carcinoma cells using nanocarriers directed to tumors is known as targeted therapy, and it provides a more specific therapy against cancer cells at the molecular level. Therefore, the use of nanocarriers is an advantageous therapeutic approach when compared to other cancer therapeutics. Over the years, it has become an enabling technology with potential for personalized cancer therapy, in which cancer cell detection, diagnosis, and therapy could be tailored to each individual tumors molecular profile because specific biomarkers could be used to predict disease progression and clinical outcomes. Here, we review nanosystems containing anticancer agents that are used for metastatic breast cancer in clinical practice, clinical trials, and research orientations. We also provide an overview of biomarkers used in metastatic breast cancer allowing for improved prognosis and therapy.

We have designed a microemulsion (ME) containing Ketoprofen (KET) for anti-inflammatory effect evaluated using the rat paw edema model. The ME was prepared by adding propylene glycol (PG) loaded with 1% KET/water (3:1, w/w), to a mixture of sorbitan monooleate and polysorbate 80 (47.0%) at 3:1 (w/w) and canola oil (38.0%). The physicochemical characterization of KET-loaded ME involved particle size and zeta potential determination, entrapment efficiency, calorimetric analysis, and in vitro drug release. The in vivo anti-inflammatory study employed male Wistar rats. Measurement of the foot volume was performed using a caliper immediately before and 2, 4, and 6 h after injection of Aerosil. KET-loaded ME showed particle size around 20 nm, with zeta potential at -16 mV and entrapment efficiency at 70%. Moreover, KET was converted to the amorphous state when loaded in the formulation and it was shown that the drug was slowly released from the ME. Finally, the in vivo biological activity was similar to that of the commercial gel, but ME better controlled edema at 4 h. These results demonstrated that the ME formulation is an alternative strategy for improving KET skin permeation for anti-inflammatory effect. Furthermore, our findings are promising considering that the developed ME was loaded with only 1% KET, and the formulation was able to keep a similar release profile and in vivo effect compared to the commercial gel with 2.5% KET. Therefore, the KET-loaded developed herein ME is likely to have a decreased side effect compared with that of the commercial gel, but both presented the same efficacy.

The use of the classic aromatic antiepileptic drugs (AAEDs) has recently been expanded to a broad spectrum of psychiatric and neurological disorders. However, the clinical use of these drugs is limited by several adverse effects, mainly idiosyncratic hepatotoxicity. AAED-induced hepatotoxicity has been attributed to a defective detoxification by the epoxide hydrolase and accumulation of arene oxides. The underlying mechanism has been proposed as immune-mediated, but direct toxicity has also been suggested. In general, idiosyncratic drug-induced hepatotoxicity may be mediated, at least in part, by oxidative stress. On the other hand, the oxidative stress induced by the AAED metabolites has not been demonstrated yet. Therefore, in the present study we have evaluated the induction of oxidative stress by three classical AAEDs: carbamazepine, phenytoin and phenobarbital as well as by their metabolites. The toxic effects of the metabolites were evaluated by incubating the drug with rat liver microsomes. The AAED-induced oxidative stress was demonstrated by the increased malondialdehyde levels, oxidation of cardiolipin; oxidation of sulfhydryl proteins and alteration of the cellular redox status. Results suggest that the hepatotoxicity associated with AAED might be mediated by the oxidative stress induced by the drugs metabolites.

Topical photodynamic therapy is used for the prevention and treatment of non-melanoma skin cancer. Until recently, clinically approved indications have been restricted to superficial basal cell carcinoma and nodular, actinic keratoses and, since 2006, Bowen’s disease. Most photosensitizers are relatively hydrophobic and will be attracted to membranes, but even the exception molecules, having substituents that render them watersoluble, bind to membranes because of their hydrophobic ring systems. The primary function of the skin, however, is to protect the body from unwanted influences from the environment. This protection is provided primarily by the stratum corneum, which consists of corneocytes surrounded by lipid regions. The major limitation of PDT is the poor penetration of photosensitizers through biological barriers, like the skin. Over the past 10 years, a considerable number of studies have therefore been conducted on the development of different strategies to overcome these difficulties, including nanocarriers to delivery photosensitizers and their precursors, nanoemulsions, liposomes, ethosomes, invasomes, liquid crystals and magnetic nanoparticles, among others.

We developed a nanodispersion system using Zinc Phtalocyanine Tetrasulphonet for photodynamic therapy. The characterization was performed based on liquid crystalline phases and photostability. We conclude that the method for preparation of nanodispersion was accomplished successfully.

PDT has been used in the treatment of malignant brain tumors for the last 2 decades. It is based on the interaction of a photosensitizer (PS) and light of an appropriate wavelength, with generation of oxygen species, mainly singlet oxygen. Brain is particularly susceptible to oxidative stress; therefore the study of PDT effects on cerebral mitochondria might provide mechanistic insights into the action of the therapy, contributing to its optimization. In the present study, we addressed the mitochondrial toxicity of the second generation PS, zinc phthalocyanine tetrasulfonate (ZnPcS4), on rat brain isolated mitochondria, by investigating both intrinsic toxicity and photodynamic action. At higher concentrations (15 and 25 microM/mg protein) ZnPcS4 caused (a) inhibition of state-3 respiration and (b) decrease of RCR and ADP/O. Electrochemical potential, state-4 respiration and Ca2+ retention capacity were not affected. Cytochrome c release was not observed. Coupled with 600 or 1800 mJ/cm2 laser irradiation, ZnPcS4 (5 microM/mg protein) caused more intense effects on state 3, RCR and ADP/O; moreover state-4 respiration and membrane potential were affected. Besides that, Ca2+ and cytochrome c release were induced. Cyclosporine A (CsA) decreased Ca2+ release and ameliorated the electrochemical potential, suggesting that membrane permeability transition (MPT) might be involved in the photodynamic effect. The low intrinsic toxicity and the high photodynamic effect on rat brain mitochondria induced by ZnPcS4, allied to its improved photophysical properties, might indicate its potential for the treatment of malignant brain tumors.

O fígado desempenha um papel central na disposição metabólica de vários agentes químicos endógenos e exógenos, incluindo quase todos os fármacos. Neste processo de biotransformação pode ocorrer a formação de metabólitos intermediários altamente reativos que se não forem convenientemente eliminados podem interagir com macromoléculas celulares lesando o órgão. A hepatotoxicidade idiossincrática associada ao uso de antiepilépticos aromáticos (AEA) é bem conhecida e tem sido atribuída ao acúmulo de intermediários tóxicos (areno-óxidos) formados durante a bioativação hepática. Embora a participação de processos imunológicos no mecanismo de ação tóxica dos AEA tenha sido demonstrada, existe a possibilidade de mecanismos adjuvantes envolvendo a toxicidade mitocondrial, evento ainda não explorado na literatura científica. Neste estudo avaliou-se, in vitro, o efeito dos AEA: carbamazepina, fenitoína e fenobarbital, bem como dos seus respectivos metabólitos na função mitocondrial e na indução do estresse oxidativo em mitocôndrias de fígado de rato, como possível mecanismo de ação hepatotóxica desses fármacos. Sistema microssomal hepático de rato foi utilizado para bioativação dos fármacos e produção dos respectivos metabólitos in vitro. Sem a bioativação, somente o fenobarbital (em concentrações elevadas) apresentou efeitos inibidores sobre o estado 3 da respiração, síntese de ATP e potencial de membrana, sem, contudo induzir o estresse oxidativo. Quando bioativados, todos os fármacos apresentaram efeitos sobre a função mitocondrial através de processo mediado por estresse oxidativo. Todos os fármacos bioativados afetaram a respiração mitocondrial, causando diminuição do consumo de oxigênio no estado 3, diminuição do RCR e aumento do consumo de oxigênio no estado 4. Foram também evidenciadas alterações na captação/liberação de cálcio, inibição da síntese de ATP, diminuição do potencial de membrana e inibição do intumescimento mitocondrial induzido pelo cálcio. A oxidação de proteínas e lipídeos mitocondriais foi demonstrada pela formação de proteínas carboniladas, diminuição de proteínas com grupamentos sulfidrila, aumento de malondialdeído (MDA) e pela oxidação da cardiolipina. O sistema de defesa antioxidante mitocondrial também foi afetado, como evidenciado pela diminuição da relação GSH/GSSG (glutationa reduzida/glutationa oxidada). Os resultados sugerem fortemente a participação do dano mitocondrial, mediado pelo estresse oxidativo causado pelos metabólitos dos AEA, no desenvolvimento da hepatotoxicidade idiossincrática induzida por esses fármacos. The liver plays a central role in the metabolic disposition of various endogenous and exogenous chemicals, including almost all drugs. During the biotransformation process, highly reactive metabolites can be produced, and if they are not detoxified, they can interact with cellular macromolecules and cause organ injury. Idiosyncratic hepatotoxicity is a well-known complication associated with aromatic antiepileptic drugs (AAED), and it has been suggested to occur due to the accumulation of toxic arene oxide metabolites. Although the participation of an immune process in the toxic action mechanism of AAED has been demonstrated, adjuvant mechanisms involving mitochondrial toxicity is also possible and such event has not been studied yet. Therefore, we investigated, in vitro, the effects of carbamazepine (CB), phenytoin (PT), phenobarbital (PB) and their respective metabolites on the hepatic mitochondrial function as well as their ability to induce oxidative stress in rat liver mitochondria, as a possible hepatotoxic action mechanism. The murine hepatic microsomal system was used to bioactivate the drugs and to produce the anticonvulsant metabolites in vitro. As an unaltered drug, only phenobarbital (in high concentrations) presented inhibitory effects on state 3 respiration, ATP synthesis, and membrane potential; however, it did not induce oxidative stress. All the bioactivated drugs affected mitochondrial function through an oxidative stress-mediated process. All the bioactivated drugs affected mitochondrial function causing decrease in state 3 respiration, decrease in RCR and increase in state 4 respiration. They also caused impairment of Ca+2 uptake /release, decrease in ATP synthesis, decrease in membrane potential and inhibition of calcium-induced swelling. Oxidation of proteins and lipids was evidenced by carbonil proteins formation, decrease in thiol proteins, increase in malonaldehyde (MDA) and cardiolipin oxidation. The mitochondrial antioxidant defense system was also affected, as evidenced by the decreased GSH/GSSG ratio (reduced glutathione/oxidized glutathione). Results strongly suggest the involvement of mitochondrial damage, which is mediated by the oxidative stress caused by the AAED metabolites, in the development of AAED-induced idiosyncratic hepatotoxicity.

Idiosyncratic hepatotoxicity is a well-known complication associated with aromatic antiepileptic drugs (AAED), and it has been suggested to occur due to the accumulation of toxic arene oxide metabolites. Although there is clear evidence of the participation of an immune process, a direct toxic effect involving mitochondria dysfunction is also possible. The effects of AAED on mitochondrial function have not been studied yet. Therefore, we investigated, in vitro, the cytotoxic mechanism of carbamazepine (CB), phenytoin (PT) and phenobarbital (PB), unaltered and bioactivated, in the hepatic mitochondrial function. The murine hepatic microsomal system was used to produce the anticonvulsant metabolites. All the bioactivated drugs (CB-B, PB-B, PT-B) affected mitochondrial function causing decrease in state three respiration, RCR, ATP synthesis and membrane potential, increase in state four respiration as well as impairment of Ca2+ uptake/release and inhibition of calcium-induced swelling. As an unaltered drug, only PB, was able to affect mitochondrial respiration (except state four respiration) ATP synthesis and membrane potential; however, Ca2+ uptake/release as well as swelling induction were not affected. The potential to induce mitochondrial dysfunction was PT-B>PB-B>CB-B>PB. Results suggest the involvement of mitochondrial toxicity in the pathogenesis of AAED-induced hepatotoxicity.