Composição farmacêutica compreendendo achyrobichalcona e uso do composto achyrobichalcona

Universidade Federal do Rio Grande do SulCiências Básicas da SaúdeFarmáciaDepositad

Physicochemical properties evaluation of the thalidomide raw material and polymorphic relationship with dissolution and compaction

O objetivo deste trabalho foi avaliar as propriedades físico-químicas de diferentes matérias-primas de talidomida visando à identificação de parâmetros críticos a serem observados para garantir a aquisição de insumo com qualidade farmacêutica. Sete amostras foram caracterizadas a partir de estudos tecnológicos, espectroscópicos, morfológicos, térmicos e cristalográficos, sendo avaliadas frente a processos de dissolução e compactação. Os resultados obtidos demonstraram a inexistência de homogeneidade entre as matérias-primas analisadas, apresentando diferenças em relação à constituição cristalina e morfológica, comportamento térmico, velocidade de dissolução intrínseca e comportamento frente à compactação. Foi observada e quantificada a presença de fases polimórficas a e b e materiais semicristalinos. A amostra T5 demonstrou desfavoráveis propriedades tecnológicas, gerando indicativos de problemas relativos à processabilidade do fármaco em medicamento, ao contrário de T1. A análise térmica forneceu indícios de transição sólido-sólido entre fases polimórficas de fármaco, provavelmente relacionada a propriedades químico-mecânicas das amostras associadas à inserção de calor. Amostras semicristalinas apresentaram maior velocidade de dissolução intrínseca, principalmente associada ao polimorfo b, representando, possivelmente, um desvio de qualidade em processos sintéticos. Estudos preliminares indicaram que a amostra T1 exibiu melhor compactabilidade que T5. Devido a sua pureza cristalográfica e propriedades tecnológicas, a amostra T1 se mostrou a mais indicada ao desenvolvimento de comprimidos de talidomida.The objective of this work was to evaluate physicochemical properties of different thalidomide raw material in order to identify critical parameters to assure the acquisition of a product with pharmaceutical quality. Seven samples were characterized by technological, spectroscopic, morphological, thermal and crystallographic approaches. Dissolution tests and degree of compaction were used to evaluate the samples. The results demonstrated raw materials with lack of homogeneity and differences related to crystal and morphological constitution, thermal behavior, intrinsic dissolution rate and compaction behavior. Polymorphic forms a and b and semicrystalline materials were observed and quantified. In contrast to T1, the sample T5 presented unfavorable technological properties generating indicatives of relative problems impairing the tablets manufacturing process. Thermal analysis indicated solid-solid transition between polymorphic phases probably related to chemical-mechanical properties of the samples associated with heating. Semicrystalline materials presented higher intrinsic dissolution rate than other samples, mainly related to the polymorph b, possibly representing quality deviation associated with synthetic process. Early studies indicated better compactability of sample T1 than T5. Due its crystallographic purity and technological properties, it is suggested that T1 is the sample choice to be used during the development of a pharmaceutical solid oral dosage form containing thalidomide

Achyrobichalcone from Achyrocline satureioides (Lam.) DC. Asteraceae : isolation, characterization, cyclodextrin complexation and evaluation of anticancer activity

Achyrobichalcona (ACB), objeto de estudo desta tese, é um biflavonoide recentemente encontrado em Achyrocline satureioides. O objetivo inicial deste trabalho voltou-se ao isolamento da ACB a partir de um extrato hidroalcoólico de A. satureioides. Para tanto, um método semi-preparativo utilizando cromatografia em contracorrente de alta velocidade foi desenvolvido. O isolamento foi realizado em duas etapas: 1) obtenção de frações enriquecidas em ACB provenientes do fracionamento do extrato; e 2) isolamento da ACB a partir destas frações enriquecidas. A separação foi realizada utilizando-se um sistema bifásico de solventes: hexano-acetato de etila-metanol-água, nas proporções 0.8:1:0.8:1, (v/v) para a primeira etapa e 0.9:0.9:0.8:1, (v/v) para a segunda etapa. O produto resultante foi purificado, cristalizado, identificado e o seu estado sólido caracterizado. O método resultou na obtenção de ACB com bom rendimento (67%) e pureza (próximo à 90%) para uso em estudos posteriores. Um método indicativo de estabilidade por cromatografia líquida de alta eficiência foi desenvolvido com o objetivo de avaliar todos os lotes de ACB isolada. Os parâmetros cromatográficos foram otimizados com o auxílio do desenho experimental Box- Behnken, gerando o sistema cromatográfico ideal para a quantificação da ACB: 37% de acetonitrila na fase móvel (63% de água/ácido ortofosfórico 1%), fluxo da fase móvel de 1,2 mL/min e 33°C de temperatura da coluna. O estudo revelou que a resolução entre impurezas e o pico da ACB e o fator de retenção foram significativamente alterados quanto os parâmetros cromatográficos foram variados. O modelo mostrou-se adequado e com boa capacidade preditiva. O método foi validado com sucesso, sendo linear, específico, exato e preciso. A robustez foi avaliada pelo desenho Plackett-Burman, demonstrando que o comprimento de onda de detecção e fluxo da fase móvel devem ser rigorosamente controlados, pois influenciam na concentração da ACB. A ACB foi instável apenas em meio alcalino; sua reação de degradação obedeceu a uma cinética de segunda ordem. A etapa seguinte objetivou avaliar comparativamente as propriedades antitumorais apresentadas por ACB, A. satureioides e outros flavonoides presentes na planta: quercetina (QCT), luteolina (LUT) e 3-O-metilquercetina (3OMQ). Primeiramente, uma revisão sobre esse tema foi elaborada com o objetivo de compilar as principais informações descritas na literatura a respeito das atividades quimioterapêuticas demonstradas para QCT, LUT, 3OMQ e A. satureioides. Subsequentemente, desenvolveu-se um estudo experimental para avaliar as propriedades antitumorais de um extrato de A. satureioides (EXT) e de seus principais flavonoides, isoladamente ou combinados na mesma proporção encontrada no EXT, frente à A549, uma linhagem de câncer de pulmão de nãopequenas células. A citotoxicidade do complexo ACB:hidroxipropil-β-ciclodextrina também foi avaliada para a A549. Os resultados demonstraram que tanto EXT (IC50%: 49 μg/mL) quanto seus flavonoides foram muito citotóxicos para a A549 (IC50%: 3OMQ-7,3 μM>ACB-13,5 μM>LUT-14,6 μM>QCT-51,1 μM), especialmente 3OMQ e ACB, avaliadas pela primeira vez em uma linhagem de câncer de pulmão. O estudo revelou que os flavonoides foram as únicas substâncias responsáveis pela citotoxicidade do extrato. A complexação entre ACB:HPβCD resultou em um grande incremento na solubilidade aquosa da ACB e aumento na citotoxicidade frente à A549 (IC50%: 9,15 μM). Todas as amostras causaram morte celular por indução da apoptose, sendo que para algumas delas a via extrínseca foi ativada (3OMQ, ACB e EXT). Sendo assim, o estudo demonstrou que A. satureioides e seus flavonoides são promissores agentes antitumorais e que a complexação com ciclodextrinas pode representar uma estratégia tecnológica importante no intuito de melhorar as propriedades físico-químicas e biológicas de moléculas bioativas.Achyrobichalcone (ACB), object of study of this thesis, is a biflavonoid recently found in Achyrocline satureioides. The initial objective of this work aimed at ACB isolation from an A. satureioides hydroalcoholic extract. For this purpose, a semi-preparative method by high-speed countercurrent chromatography (HSCCC) was developed. The isolation was achieved in two steps: 1) obtaining ACB enriched fractions from the fractioning of the extract; and 2) ACB separation from these enriched fractions. The separation was obtained using a biphasic solvent system: hexane-ethyl acetatemethanol- water, at proportions 0.8:1:0.8:1 (v/v) for the first step and 0.9:0.9:0.8:1 (v/v) for the second step. The resulting product was purified, crystallized, identified and the solid state was characterized. The method resulted in ACB in good yield (67%) and purity (about 90%) for use in further studies. A stability-indicating high performance liquid chromatographic method was developed in order to evaluate all batches of ACB isolated. The chromatographic parameters were optimized by Box- Behnken design, providing the ideal chromatographic system for ACB quantification: 37% of acetonitrile on the mobile phase (63% water/orthophosphoric acid 1%), flow rate of the mobile phase of 1.2 ml/min and 33°C of column temperature. The resolution between impurities and ACB peak and retention factor were significantly changed when chromatographic parameters were altered. The model was suitable, with good predictive capacity. The method was successful validated, being linear, specific, accurate and precise. Plackett-Burman robustness test demonstrated that detection wavelength and flow rate of mobile phase must be strictly controlled because influenced ACB concentration. ACB was unstable only in alkaline medium, and the degradation reaction was guided by a second-order kinetics. The next step aimed a comparative evaluation of the anticancer properties of ACB, A. satureioides extract and others flavonoids present in the plant: quercetin (QCT), luteolin (LUT) and 3-O-methylquercetin (3OMQ). Firstly, a review of this theme was performed to compile the main information described in literature about chemotherapeutics activities of QCT, LUT, 3OMQ and A. satureioides. Subsequently, an experimental study was carried out to evaluate the anticancer properties of an A. satureioides extract (EXT) and its major flavonoids alone or combined in the same proportion found in EXT, against A549, a non-small cancer lung cell line (NSCLC). In addition, the cytotoxicity of a ACB:hydroxipropil-β-cyclodextrin (ACB:HPβCD) complex was also assessed for A549. The results showed that both EXT (49 μg/mL) as its flavonoids were highly cytotoxic for A549 (GI50%: 3OMQ-7.3 μM>ACB-13.5 μM>LUT-14.6 μM>QCT-51.1 μM), especially 3OMQ and ACB, evaluated for the first time against lung cancer. The study showed that flavonoids were the unique substances responsible for the cytotoxicity of the extract. ACB:HPβCD complex allowed a great enhance in water solubility of ACB and cytotoxicity against A549 (GI50%: 9.15 μM). All the samples induced cell death by apoptosis induction, and for some of them the extrinsic pathway was activated (3OMQ, ACB and EXT). Taken together, the study demonstrated that A. satureioides and its derived flavonoids are promising anticancer agents and, additionally, the cyclodextrin complexation can represent an important technological strategy for improve physicochemical and biological properties of bioative molecules

Composição farmacêutica compreendendo achyrobichalcona e uso do composto achyrobichalcona

Universidade Federal do Rio Grande do SulCiências Básicas da SaúdeFarmáciaDepositad

Supplementation with Achyrocline satureioides inflorescence extracts to pregnant and breastfeeding rats induces tissue-specific changes in enzymatic activity and lower neonatal survival

Achyrocline satureioides (AS, family Asteraceae) is a plant widely used in traditional medicine for stomach, digestive, and gastrointestinal disorders during pregnancy. Studies regarding the indiscriminate use of plant infusions during pregnancy are limited. Recent reports have shown that chronic flavonoid supplementation induces toxicity in vivo and raises the mortality rates of healthy subjects. Therefore, we investigated whether supplementation of pregnant and lactating Wistar rats with two AS inflorescence extracts, consisting of an aqueous (AQ) extract similar to a tea (47 mg kg1 day) and a hydroethanolic (HA) extract (35 mg kg1 day1) with a higher flavonoid content, could induce redox-related side effects. Total reactive antioxidant potential (TRAP), thiobarbituric reactive species (TBARS), and total reduced thiol (SH) content were evaluated. Superoxide dismutase (SOD) and catalase (CAT) activities were additionally quantified. Our data suggest that both AQ and HA of AS inflorescence extracts may induce symptoms of toxicity in concentrations of (47 mg kg1 day) and (35 mg kg1 day1), respectively, in mothers regarding the delivery index and further decrease of neonatal survival. Of note, significant tissue-specific changes in maternal (liver, kidney, heart, and hippocampus) and pups (liver and kidney) biochemical oxidative parameters were observed. Our findings provide evidence that may support the need to control supplementation with the AQ of AS inflorescence extracts during gestation due to potential toxicity in vivo, which might be related, at least in part, to changes in tissue-specific redox homeostasis and enzymatic activity

Supplementation with Achyrocline satureioides Inflorescence Extracts to Pregnant and Breastfeeding Rats Induces Tissue-Specific Changes in Enzymatic Activity and Lower Neonatal Survival

Achyrocline satureioides (AS, family Asteraceae) is a plant widely used in traditional medicine for stomach, digestive, and gastrointestinal disorders during pregnancy. Studies regarding the indiscriminate use of plant infusions during pregnancy are limited. Recent reports have shown that chronic flavonoid supplementation induces toxicity in vivo and raises the mortality rates of healthy subjects. Therefore, we investigated whether supplementation of pregnant and lactating Wistar rats with two AS inflorescence extracts, consisting of an aqueous (AQ) extract similar to a tea (47 mg·kg−1·day) and a hydroethanolic (HA) extract (35 mg·kg−1·day−1) with a higher flavonoid content, could induce redox-related side effects. Total reactive antioxidant potential (TRAP), thiobarbituric reactive species (TBARS), and total reduced thiol (SH) content were evaluated. Superoxide dismutase (SOD) and catalase (CAT) activities were additionally quantified. Our data suggest that both AQ and HA of AS inflorescence extracts may induce symptoms of toxicity in concentrations of (47 mg·kg−1·day) and (35 mg·kg−1·day−1), respectively, in mothers regarding the delivery index and further decrease of neonatal survival. Of note, significant tissue-specific changes in maternal (liver, kidney, heart, and hippocampus) and pups (liver and kidney) biochemical oxidative parameters were observed. Our findings provide evidence that may support the need to control supplementation with the AQ of AS inflorescence extracts during gestation due to potential toxicity in vivo, which might be related, at least in part, to changes in tissue-specific redox homeostasis and enzymatic activity

Supplementation with Achyrocline satureioides inflorescence extracts to pregnant and breastfeeding rats induces tissue-specific changes in enzymatic activity and lower neonatal survival

Achyrocline satureioides (AS, family Asteraceae) is a plant widely used in traditional medicine for stomach, digestive, and gastrointestinal disorders during pregnancy. Studies regarding the indiscriminate use of plant infusions during pregnancy are limited. Recent reports have shown that chronic flavonoid supplementation induces toxicity in vivo and raises the mortality rates of healthy subjects. Therefore, we investigated whether supplementation of pregnant and lactating Wistar rats with two AS inflorescence extracts, consisting of an aqueous (AQ) extract similar to a tea (47 mg kg1 day) and a hydroethanolic (HA) extract (35 mg kg1 day1) with a higher flavonoid content, could induce redox-related side effects. Total reactive antioxidant potential (TRAP), thiobarbituric reactive species (TBARS), and total reduced thiol (SH) content were evaluated. Superoxide dismutase (SOD) and catalase (CAT) activities were additionally quantified. Our data suggest that both AQ and HA of AS inflorescence extracts may induce symptoms of toxicity in concentrations of (47 mg kg1 day) and (35 mg kg1 day1), respectively, in mothers regarding the delivery index and further decrease of neonatal survival. Of note, significant tissue-specific changes in maternal (liver, kidney, heart, and hippocampus) and pups (liver and kidney) biochemical oxidative parameters were observed. Our findings provide evidence that may support the need to control supplementation with the AQ of AS inflorescence extracts during gestation due to potential toxicity in vivo, which might be related, at least in part, to changes in tissue-specific redox homeostasis and enzymatic activity