9 research outputs found
Formulações responsivas ao ambiente contendo óleo resina de Copaifera reticulata Duche no combate à leishmaniose cutânea vísceral : indução da resposta imune TH1 e perfil de segurança
No full textOrientador: Prof. Dr. Celso Vataru NakamuraCoorientador: Prof.ª Dr.ª Danielle Lazarin BidóiaTese (doutorado em Ciências Biológicas) - Universidade Estadual de Maringá, 2024RESUMO: As leishmanioses representam um desafio global em saúde pública devido à falta de avanços significativos no desenvolvimento de novas terapias desde a introdução dos antimoniais pentavalentes na década de 1940. Diante da limitada eficácia terapêutica e da alta toxicidade dos medicamentos disponíveis, a investigação de novas substâncias, baseada em conhecimentos empíricos e no uso tradicional de plantas medicinais, tem se mostrado uma alternativa promissora. Dentre as espécies vegetais de interesse, as pertencentes ao gênero Copaifera se destacam, sendo o óleo resina extraído dessas plantas um recurso natural de grande importância na medicina tradicional da região Amazônica, com atividade antileishmania promissora. No entanto, para garantir a qualidade, segurança e a eficácia dos fitoterápicos, é fundamental realizar o controle de qualidade desses produtos, desde a identificação da espécie vegetal até a presença dos agentes bioativos na formulação final, assim como garantir a sua biodisponibilidade. Além disso, o desenvolvimento de formulações farmacêuticas requer várias etapas para garantir a estabilidade do medicamento, a preservação da atividade das substâncias e a melhoria da solubilidade e biodistribuição. Nesse contexto, com base no uso tradicional do óleo de copaíba (OC) e nas evidências de sua segurança, o presente estudo teve como objetivo avaliar a eficácia terapêutica de um sistema auto emulsionante (F14) para administração oral e de um sistema bioadesivo termoresponsivo (F12) tópico para o tratamento da leishmaniose cutânea e visceral. Foram conduzidos estudos in vivo utilizando camundongos infectados ou não com Leishmania amazonensis ou L. infantum, nos quais foram administrados o óleo resina de C. reticulata Ducke isolado ou incorporado nas formulações F14 e F12. Os efeitos dos tratamentos foram avaliados por meio de medidas de peso corporal e dos órgãos, determinação do volume das lesões e carga parasitária utilizando técnicas de bioluminescência e qPCR, avaliação dos níveis de citocinas e produção de oxido nítrico, além de análises bioquímicas e hematológicas para detectar possíveis efeitos tóxicos. Os resultados indicaram que os animais não infectados submetidos aos tratamentos com o OC e as formulações não apresentaram alterações significativas no peso corporal nem nos pesos dos órgãos, sugerindo a ausência de toxicidade dessas terapias. No entanto, nos animais infectados com L. infantum, observou-se uma redução significativa no peso corporal quando tratados com a formulação F14 com OC ou miltefosina, apesar de não apresentarem diferenças na aparência física. Essa diminuição no peso corporal foi acompanhada por uma redução no peso do fígado em todos os grupos de tratamento em comparação com o grupo controle, indicando uma possível reversão da hepatomegalia, uma
característica da leishmaniose visceral, como resultado dos tratamentos administrados. Essas reduções nos pesos dos órgãos alvo foram então atribuídas à diminuição da carga parasitária confirmada por técnicas de qPCR, demonstrando a eficácia das intervenções terapêuticas. No modelo experimental de infecção com L. amazonensis, não foram observadas variações significativas no peso corporal ou nos órgãos após a necropsia. Ao avaliar o aumento no volume das lesões nas patas desses camundongos após 28 dias de tratamento, constatou-se uma redução de 64,25% e 45,42% para a formulação F14 com OC e a formulação F12 com OC, respectivamente, em comparação com o grupo controle. Esses resultados foram corroborados pelas medidas de bioluminescência e qPCR, que indicaram que a redução no volume das lesões foi diretamente proporcional a redução na carga parasitária. A ação imunomoduladora do OC e das formulações também foi avaliado, sendo observado uma polarização da resposta Th1 com aumentos nos níveis de IL-2, TNF-? e IFN-? concomitante com a redução de IL-10 e TGF-?, que como consequência levaram a aumentos de oxido nítrico e morte dos parasitos. Além da eficácia terapêutica, a segurança do uso das formulações F14 e F12 com OC foi avaliada, e os resultados indicam que essas formulações não causaram toxicidade significativa nos animais, uma vez que os marcadores renais e hepáticos apresentaram valores semelhantes aos observados nos animais não tratados. Apesar dos estudos toxicológicos publicados demonstrarem que o óleo resina de Copaifera spp. não possui efeitos mutagênico, genotóxico ou citotóxicos, e os polímeros e emulsionantes utilizados no desenvolvimento das formulações F14 e F12 serem reconhecidos como seguros e aprovados para uso em formulações farmacêuticas, ainda eram necessários estudos de toxicidade de ambas as formulações. Uma vez que a identificação de qualquer atividade toxicológica em substâncias bioativas de plantas por meio de testes pré-clínicos deve preceder seu uso em estratégias de saúde preventiva. Sendo assim, a toxicidade aguda das formulações foi avaliada em ratos Wistar machos e fêmeas ao longo de um período de 14 dias. Os ratos receberam uma dose única da formulação F14 com OC por via oral ou intraperitoneal ou da formulação F12 com OC por via tópica, na dose de 2.000 mg/kg. Avaliamos a toxicidade comportamental, bem como as alterações bioquímicas e hematológicas, incluindo toxicidades em múltiplos órgãos. A administração das formulações não resultou em mortalidade dos ratos, e nenhum efeito toxicológico foi observado em relação ao peso corporal, consumo de ração e água, peso dos órgãos ou parâmetros hematológicos e bioquímicos. Complementarmente, camundongos Swiss foram tratados com as formulações na dose de 2.000 mg/kg por 24 h e, então, realizada a coleta de medula óssea para realizar o teste de micronúcleos para avaliar potencial atividade genotóxica das formulações, a qual não foi
evidenciada. Em conclusão, neste trabalho, descrevemos a eficácia terapêutica e segurança do uso das formulações F14 e F12 contendo óleo resina de C. reticulata Ducke como alternativa terapêutica para o tratamento da leishmaniose cutânea e visceral. Com estas abordagens pretende-se obter os dados necessários para notificação de novo produto fitoterápico de acordo com o estipulado pela Agência Nacional de Vigilância Sanitária (ANVISA), Food and Drug Administration (FDA - Estados Unidos da América) e demais agências reguladoras como uma alternativa de baixo custo para o tratamento de leishmaniosesABSTRACT: Leishmaniasis presents a global public health challenge due to the lack of significant advancements in developing new therapies since the introduction of pentavalent antimonials in the 1940s. Given the limited therapeutic efficacy and high toxicity of available medications, investigating new substances based on empirical knowledge and the traditional use of medicinal plants has emerged as a promising alternative. Among the plant species of interest, those belonging to the genus Copaifera are notable. The oil-resin extracted from these plants is a natural resource of great importance in traditional medicine in the Amazon region, exhibiting promising anti-leishmanial activity. However, to ensure the quality, safety, and efficacy of phytotherapeutics, it is essential to conduct quality control of these products, from the identification of the plant species to the presence of bioactive agents in the final formulation, as well as to guarantee their bioavailability. Additionally, developing pharmaceutical formulations requires several steps to ensure the drug's stability, preservation of the active substances' activity, and improvement of solubility and biodistribution. In this context, based on the traditional use of copaiba oil (CO) and evidence of its safety, this study aimed to evaluate the therapeutic efficacy of a self-emulsifying system (F14) for oral administration and a topical thermoresponsive bioadhesive system (F12) for treating cutaneous and visceral leishmaniasis. In vivo studies were conducted using mice infected or not with Leishmania amazonensis or L. infantum, where C. reticulata Ducke oil-resin was administered either in isolation or incorporated into the F14 and F12 formulations. The treatment effects were evaluated by measuring body and organ weights, determining lesion volume and parasitic load using bioluminescence and qPCR techniques, assessing cytokine levels and nitric oxide production, and conducting biochemical and hematological analyses to detect potential toxic effects. The results indicated that uninfected animals subjected to CO and formulation treatments did not exhibit significant changes in body weight or organ weights, suggesting the absence of toxicity of these therapies. However, in animals infected with L. infantum, a significant reduction in body weight was observed when treated with the F14 formulation with CO or miltefosine, despite no differences in physical appearance. This decrease in body weight was accompanied by a reduction in liver weight in all treatment groups compared to the control group, indicating a possible reversal of hepatomegaly, a characteristic of visceral leishmaniasis, as a result of the administered treatments. These reductions in target organ weights were attributed to the decreased parasitic load confirmed by qPCR techniques, demonstrating the efficacy of the therapeutic interventions. In the experimental infection model with L. amazonensis, no
significant variations in body or organ weights were observed after necropsy. When evaluating the increase in lesion volume on the paws of these mice after 28 days of treatment, reductions of 64.25% and 45.42% were observed for the F14 formulation with CO and the F12 formulation with CO, respectively, compared to the control group. These findings were corroborated by bioluminescence and qPCR measurements, which indicated that the reduction in lesion volume was directly proportional to the reduction in parasitic load. The immunomodulatory action of CO and the formulations was also evaluated, revealing a Th1 response polarization with increased levels of IL-2, TNF-?, and IFN-?, concomitant with reduced levels of IL-10 and TGF-?, leading to increased nitric oxide production and parasite death. Besides therapeutic efficacy, the safety of using the F14 and F12 formulations with CO was evaluated, and the results indicate that these formulations did not cause significant toxicity in the animals, as renal and hepatic markers showed values similar to those observed in untreated animals. Despite published toxicological studies demonstrating that Copaifera spp. oil-resin does not possess mutagenic, genotoxic, or cytotoxic effects, and the polymers and emulsifiers used in developing the F14 and F12 formulations are recognized as safe and approved for use in pharmaceutical formulations, toxicity studies of both formulations were still necessary. Identifying any toxicological activity in bioactive plant substances through preclinical tests must precede their use in preventive health strategies. Thus, the acute toxicity of the formulations was evaluated in male and female Wistar rats over 14 days. The rats received a single dose of the F14 formulation with CO orally or intraperitoneally or the F12 formulation with CO topically at a dose of 2,000 mg/kg. We assessed behavioral toxicity and biochemical and hematological changes, including multi-organ toxicities. The administration of the formulations did not result in rat mortality, and no toxicological effects were observed regarding body weight, food and water consumption, organ weights, or hematological and biochemical parameters. Additionally, Swiss mice were treated with the formulations at a dose of 2,000 mg/kg for 24 h, and bone marrow was collected for micronucleus testing to assess potential genotoxic activity of the formulations, which was not evidenced. In conclusion, this work describes the therapeutic efficacy and safety of the F14 and F12 formulations containing C. reticulata Ducke oil-resin as an alternative therapy for cutaneous and visceral leishmaniasis. These approaches aim to obtain the necessary data for the notification of a new phytotherapeutic product in accordance with the regulations of the National Health Surveillance Agency (ANVISA), the Food and Drug Administration (FDA - United States of America), and other regulatory agencies as a low-cost alternative for leishmaniasis treatment100 f. : il. (algumas color.)
Development of Environmentally Responsive Self-Emulsifying System Containing Copaiba Oil-Resin for Leishmaniasis Oral Treatment
No full textLeishmaniasis is a disease caused by protozoa species of the Leishmania genus, and the current treatments face several difficulties and obstacles. Most anti-leishmanial drugs are administered intravenously, showing many side effects and drug resistance. The discovery of new anti-leishmanial compounds and the development of new pharmaceutical systems for more efficient and safer treatments are necessary. Copaiba oil-resin (CO) has been shown to be a promising natural compound against leishmaniasis. However, CO displays poor aqueous solubility and bioavailability. Self-emulsifying drug delivery systems (SEDDS) can provide platforms for release of hydrophobic compounds in the gastrointestinal tract, improving their aqueous solubilization, absorption and bioavailability. Therefore, the present work aimed to develop SEDDS containing CO and Soluplus® surfactant for the oral treatment of leishmaniasis. The design of the systems was accomplished using ternary phase diagrams. Emulsification and dispersion time tests were used to investigate the emulsification process in gastric and intestinal environments. The formulations were nanostructured and improved the CO solubilization. Their in vitro antiproliferative activity against promastigote forms of L. amazonensis and L. infantum, and low in vitro cytotoxicity against macrophages were also observed. More studies are necessary to determine effectiveness of SOL in these systems, which can be candidates for further pharmacokinetics and in vivo investigations
Silk fibroin nanofibers containing chondroitin sulfate and silver sulfadiazine for wound healing treatment
No full textInternational audienceNanofibers containing silk fibroin (SF), chondroitin sulfate (CS) and silver sulfadiazine (SSD) were made up by electrospinning technique. Poly(ethylene oxide) (PEO) was added to the acidic water solution used for electrospinning to improve electrospinnability. Fibers were characterized by Scanning Electron Microscopy (SEM), Fourier transform infrared spectroscopy (FTIR-ATR), Energy Dispersive X-ray Spectroscopy (EDS), Differential Scanning Calorimetry (DSC) and biological analysis. The FTIR-ATR and DSC results showed that PEO was removed from the surface of the nanofibers by washing the as-spun fibers with absolute ethanol. FTIR-ATR and EDS results showed that CS and SSD remained in the nanofibers. Biological assays showed that the as-obtained nanofibers are not toxic to healthy Vero cells. Antibacterial activity tests showed that the concentration of CS significantly influenced the antibacterial activity of the samples. In vivo experiments with Wistar rats revealed that SF nanofibers containing CS and SSD had similar results to standard SSD cream treatment, with the advantage of being applied only once to the patient avoiding discomfort and pain in dressing changes. CS played an important role as a stabilizing agent of electrospinning solutions improving SSDremaining and enhancing the antibacterial activity of nanofibers against bacteria through in vitro
Multifunctional Nanoparticles as High-Efficient Targeted Hypericin System for Theranostic Melanoma
No full textBiotin, spermine, and folic acid were covalently linked to the F127 copolymer to obtain a new drug delivery system designed for HY-loaded PDT treatment against B(16)F(10) cells. Chemical structures and binders quantification were performed by spectroscopy and spectrophotometric techniques ((1)NMR, HABA/Avidin reagent, fluorescamine assay). Critical micelle concentration, critical micelle temperature, size, polydispersity, and zeta potential indicate the hydrophobicity of the binders can influence the physicochemical parameters. Spermine-modified micelles showed fewer changes in their physical and chemical parameters than the F127 micelles without modification. Furthermore, zeta potential measurements suggest an increase in the physical stability of these carrier systems. The phototherapeutic potential was demonstrated using hypericin-loaded formulation against B(16)F(10) cells, which shows that the combination of the binders on F127 copolymer micelles enhances the photosensitizer uptake and potentializes the photodynamic activity
Antileishmanial activity of <i>neo</i>-clerodane diterpenes from <i>Croton echioides</i>
No full textNowadays, new leishmanicidal drugs are needed and natural products arise as a promising alternative source. Therefore, bioguided fractionation of a hydroethanolic extract from the stem bark of Croton echioides Baill. were conducted based on its antileishmanial activity. Two novel neo-clerodane diterpenoids methyl-15,16-epoxy-3,13(16),14-neo-clerodatrien-17,18-dicarboxylate (1) and dimethyl-3-oxo-15,16-epoxy-13(16),14-neo-clerodadien-17,18-dicarboxylate (2) were isolated, as well as four known compounds (3–6) and lupeol, from the hexane fraction. Their structures were established by NMR analysis. The crude extract, fractions and the compounds (1 and 3–6) were evaluated for their in vitro antileishmanial activity and cytotoxicity against macrophages J774A.1. The selectivity index (SI) were calculated. The most active compound against promastigote forms of L. amazonensis was the clerodane diterpene 4, with IC50 values of 8.3 µM and SI value of 80.9. Our results highlighted stem bark of Croton echioides Baill. as a promising source for the development of a new chemotherapeutic agent to combat leishmaniasis. </p
New cadinene-sesquiterpene from <i>Chromolaena laevigata</i> (lam.) R. M. King & H. Rob (Asteraceae) aerial parts and biological activities
No full textPhytochemical investigation of Chromolaena laevigata led to the isolation of a new cadinene-sesquiterpene, chromolaevigone glucoside (1), along with nine known compounds: daucosterol (2), stigmasterol glycoside (3), stigmasterol (4), β-sitosterol (5), pilloin (6), gonzalitosin I (7), quercetin-3-O-α-rhamnopyranoside (8), 7,7-dihydroxy-calamen-12-oic acid lactone (9) and trachelanthic acid (10). Others 11 known compounds were identified by UHPLC-HRMS/MS. These compounds are being described for the first time in this species, with the exception of cadinene 9. Furthermore, due to the limitation of pharmacological studies, antiproliferative, antiviral, and antimicrobial activities of C. laevigata were evaluated. The best results in the cytotoxicity, antimicrobial and antiproliferative tests, presenting GI50 values on ovarian tumour cells (OVCAR-03) of 1.9 μg mL−1 and kidney (786-0) of 2.5 μg mL−1 were observed for the hexanic fraction. </p
Formulation and performance evaluation of emulgel platform for combined skin delivery of curcumin and propolis
No full textThe environment can modify the physiology and body protective function of the skin. Propolis (PRP) and curcumin (CUR) possess important antioxidant and antimicrobial properties, and they can be administered in a combined way and using photodynamic therapy (PDT). Emulgels can control drug release due to the physicochemical properties of the gel and the emulsion. They constitute a good strategy for achieving an improved platform for the combined delivery of PRP and CUR. There are no other studies of emulgels composed of PRP and CUR and their performance as antimicrobial and skin healing using or not PDT. This study aimed to investigate the effect of Carbopol 934 P (C934P), 974 P (C974P) or polycarbophil (PC) on physicochemical stability, antioxidant activity, drug release profile, antimicrobial activity, and ex vivo skin permeation and retention of emulgels containing PRP and CUR. Formulations containing C974P or PC displayed improved stability and antioxidant activity. They displayed activity against Staphylococcus aureus and modified (extended) drug release, governed mainly by non-Fickian anomalous transport. C974P and PC resulted in improved emulgels for combined CUR and PRP delivery, allowing the drugs to cross the stratum corneum, and permeate the epidermis, reaching the dermis. The selected emulgels are candidates for further studies to prove their action and benefits to skin health.</p
Exercício do direito coletivo à saúde na pós-modernidade jurídico-política brasileira: nova estrutura participativa para a formulação das políticas do sistema único
No full textTese (doutorado) - Universidade Federal de Santa Catarina, Centro de Ciências Jurídicas, Programa de Pós-Graduação em Direito, Florianópolis, 2013.Esta tese tem como tema o direito à saúde e, particularmente, o direito coletivo à saúde e o seu exercício por meio de uma estrutura participativa pós-moderna na fase de formulação das políticas do Sistema Único. A sua elaboração parte da constatação do cenário jurídico-político inédito inaugurado no Brasil pela Constituição de 1988 que: configurou o Estado Democrático de Direito; introduziu a República Federativa Participativa; reconheceu o direito à saúde como um direito social fundamental de todos, a ser garantido pelo Estado por meio de políticas públicas formuladas e operacionalizadas no seio do Sistema Único de Saúde (SUS), cuja diretriz constitucional da participação ?comunitária? levou à criação legal dos Conselhos como instâncias participativas competentes para deliberar sobre as políticas do Sistema. A tese ocupa-se em responder como o direito de todos à saúde pode ser exercido no âmbito do novo paradigma introduzido pela Carta Magna. O que mais chama atenção é o fato de o panorama constitucional, que inaugura um paradigma jurídico-político pós--moderno, ainda ter os seus elementos e categorias estruturados, orientados e configurados sobre fundamentos da ordem jurídico-política moderna, que não permite a compreensão da complexidade e abrangência das mudanças introduzidas. Por isso, a tese faz uma análise de cunho paradigmático, capaz não só de explicar as novas categorias constitucionais direito de todos à saúde e SUS, mas de fornecer-lhes diretrizes e bases condizentes ao seu contexto e, assim, capazes de fundamentar a concepção de formas apropriadas para o exercício participativo de tal direito quando da formulação das políticas do Sistema Único. Embora o texto constitucional tenha conduzido à criação legal dos Conselhos de Saúde como foros de deliberação ?comunitária? das políticas do SUS, o exame de cunho paradigmático ora proposto evidencia que este modelo ainda não corresponde às características da República Federativa Participativa, que exige que a saúde de todos seja reconhecida no plano jurídico como um bem coletivo autônomo exercido por meio da participação direta do seu titular, a Sociedade personalizada, em processos próprios. Desta forma, a partir de uma análise crítica do paradigma jurídico-político moderno e da compreensão do pós-moderno, de suas características e consequências para o exercício do direito coletivo à saúde, analisam-se as limitações do modelo de exercício do direito de todos à saúde por meio da formulação das políticas do SUS pelos Conselhos de Saúde, para comprovar a necessidade de outra estrutura de participação, que seja condizente com os elementos e processos coletivos próprios do novo paradigma. Partindo de tais fundamentos, a tese apresenta a idealização de uma estrutura participativa inovadora, na qual a Sociedade atua diretamente para construir as deliberações sobre as políticas do Sistema de Saúde ? que ditam o conteúdo e os rumos do direito coletivo que lhe pertence. Trata-se da concepção de um processo inclusivo, que não prescinde da estrutura administrativa estatal e de suas autoridades. Um modelo que, nas oportunidades de sua utilização, altera a configuração participativa do SUS e fornece novos papéis aos Conselhos de Saúde. Abstract : This thesis has as theme the right to health and, particularly, the collective health right and its practice through a post-modern participative structure in the phase of the Unified System?s policies formulation. Its elaboration starts with the observation of the unprecedented legal-political scenario inaugurated in Brazil by the 1998 Constitution which: configured the Democratic State of Law; introduced the Participative Federal Republic; recognized the right to health as a social fundamental right of all, to be guaranteed by the State through the public policies formulated and applied in the core of the Unified Public Health System (hereby SUS), whose constitutional guideline of the community participation led to the legal creation of Councils like participative instances competent to deliberate about the System?s policies. The thesis aims to answer how the right of all to health can be practiced in the range of the new model introduced by the Magna Carta. What receives special attention is the fact that the constitutional panorama, which inaugurates post-modern legal-political paradigm, still has its elements and categories structured, orientated and configured about fundaments of the modern legal-political order, which does not allow the comprehension of the complexity and range of the introduced changes. Therefore, the thesis does an analyses of paradigmatic character, able not only to explain the new constitutional categories right of all to health and to the Unified Public Health System (SUS), but also of giving them guidelines and basis according to its context and, as such, capable of justifying the conception of proper forms to the participative practice of such right when the policies of the Unified Public Health System are formulated. Despite the fact that the constitutional text has conducted to the legal creation of the Health Boards as forums of ?community? deliberation of the SUS policies, the exam of paradigmatic character, proposed by then, highlights that this model does not correspond to the characteristics of the Participative Federal Republic, which demands that the health of all be recognized as an autonomous collective well-being practiced through the direct participation of its holder, the Society personified, in its own lawsuits. Thus, from a critic analysis of the modern legal-political paradigm and from the comprehension of the post-modern, of its characteristics and consequences to the practice of the collective health law, the limitations of the model of the right of all to health practice through the formulation of SUS? policies by the Health Councils are analyzed, in order to prove the need of another structure of participation, which is adjusted with the elements and collective processes from the new paradigm. Starting with these fundaments, the thesis presents the idealization of an innovative participatory structure, in which the Society acts directly to build the deliberations about the Health System policies ? which dictate the content and the directions of the collective right that belongs to itself. It is about the conception of an inclusive process, which does not need the state administrative structure and its authorities. A model that, in the opportunities of its use, alters the participative configure of SUS and gives new roles to the Health Boards
