Estudo da qualidade fisico-química de comprimidos similares e genéricos de nimesulida 100 mg e validação de metodologia analítica para cápsulas magistrais

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Ciências da Saúde, Programa de Pós-Graduação em Ciências Farmacêuticas, 2014.A nimesulida é um anti-inflamatório não esteroide (AINE) inibidor seletivo da ciclo-oxigenase 2 (COX-2), classificada como um fármaco de classe II pelo Sistema de Classificação Biofarmacêutica. Existe no mercado uma grande variedade de formas farmacêuticas orais sólidas contendo nimesulida, incluindo comprimidos industrializados genéricos, similares e cápsulas magistrais. No presente trabalho foi realizada a avaliação físico-química dos comprimidos industrializados de sete diferentes laboratórios, sendo três genéricos (G1, G2 e G3), três similares (S1, S2 e S3), o medicamento referência R, além de cápsulas magistrais de cinco farmácias de manipulação de Brasília (M1-M5). O padrão de referência da nimesulida foi caracterizado como puro a partir de testes de ponto de fusão e ao comportamento espectrofotométrico na região do infravermelho e do UV/visível. Um método analítico por espectrofotometria no UV/visível para a determinação do teor de nimesulida e um ensaio de dissolução de cápsulas magistrais foram satisfatoriamente validados. Os perfis de dissolução dos medicamentos em tampão fosfato de potássio pH 7,4 com polissorbato 80 a 2% foram realizados com 12 unidades de cada produto, utilizando-se dois aparatos, 37°C, rotação de 75 RPM e coletas de 5 mL de 5 a 45 minutos. Os perfis de dissolução dos comprimidos industrializados foram comparados estatisticamente com a aplicação dos fatores de diferença e semelhança (f1 e f2), cálculo da eficiência de dissolução (ED%) e cinética de dissolução com modelos de zero ordem, quadrático e Higuchi. Todos os produtos industrializados apresentaram características físico-químicas de acordo com o preconizado pela Farmacopeia Brasileira. As cápsulas magistrais M1, M2 e M4 não apesentaram resultados satisfatórios quanto ao teor de nimesulida, e M1 também foi reprovado em relação à uniformidade de doses unitárias. Todos os medicamentos industrializados exibiram valores de dissolução de mais de 75% dentro do tempo de 45 minutos. Nenhuma das cápsulas atingiu este valor. Com exceção de S3 (f2 = 47,24), os produtos industrializados apresentaram perfis de dissolução similares a R. A ED% média de R comparado com S3, G2 e G3 foi estatisticamente diferente. O modelo Higuchi foi o mais adequado para descrever a cinética de dissolução dos medicamentos industrializados. Os resultados obtidos nesse estudo mostraram a necessidade de uma revisão da legislação atual no que diz respeito às farmácias magistrais, de maneira a garantir qualidade equivalente para todos os tipos de formulações orais sólidas disponíveis ao paciente.Nimesulide is a non-steroidal anti-inflammatory drug that selectively inhibits cyclo-oxigenase-2 (COX-2), and a class-II drug according to the Biopharmaceutical Classification System. A great variety of solid oral dosage forms that contain nimesulide is available in the market, including generics and similar medicines, and capsules prepared in magistral pharmacies. In this study, the tablets of seven different laboratories were analyzed for their physical-chemical properties, three generic products (G1, G2 and G3), three similar (S1, S2 and S3); and one reference product (R). Five capsules (M1- M5) were obtained from five different magistral pharmacies from Brasilia. The nimesulide reference standard was considered pure after the determination of melting point and spectrophotometric behavior in infrared and UV/visible regions. An UV/visible spectrophotometric analytical method to determine the content of nimesulide in magistral capsules and a dissolution test for magistral capsules were satisfactorily validated. The dissolution profiles in potassium phosphate buffer pH 7.4 with 2% polysorbate 80 of the products were determined with 12 units, at 37°C, with paddles (tablets) and baskets (capsules), 75 RPM, and 5 mL aliquots withdrawn at 5 to 45 minutes. The difference and similar factors (f1 e f2) and dissolution efficiency (DE%) and dissolution kinetics were determined by the application of zero order, quadratic and Higuchi models. Tukey t-test was used to compare the mean values of DE%. The physical-chemical properties of all tablet products were in accordance to the Brazilian Pharmacopoeia. Magistral products M1, M2 and M4 did not yield acceptable nimesulide content results, and M1 was disapproved regarding uniformity of dosage units. All tablet products showed more than 75% of dissolution within 45 minutes of process. None of the capsules reached this value. With the exception of S3 (f2 = 47,24), the dissolution profiles of tablet products were similar to product R. DE% between product R and G2, G3 and S3 were statistically different. The Higuchi model was the most suitable to describe the dissolution profiles of the tablet products. The results of this study showed the need of a revision of the Brasilian legislation concerning the magistral pharmacies in order to garantee products of good qualities of all types of formulations available to the patient

NIMESULIDE: DISSOLUTION PROFILE, VALIDATION OF ANALYTICAL METHODS FOR CAPSULES, AND ASSESSMENT OF PRODUCT QUALITY

Objective: The main objective of this paper was to evaluate the quality of similar (S, n=3) and generic (G, n=3) tablets and compounding capsules (C, n=6) containing nimesulide (100 mg).Methods: The parameters investigated (weight, nimesulide content, uniformity of dosage units, disintegration, friability and hardness (tablets) and dissolution profile) were evaluated against the Brazilian Pharmacopeia and a reference compound (for tablets). Nimesulide content, determined by a UV/visible spectrophotometric method, and dissolution test were validated for compounding capsules.Results: All formulations had a mean weight coefficient of variation lower than 5%. Three compounding formulations contained less than 95 mg nimesulide, with C1 (88.5 mg) also showing a lack of dosage unit uniformity. Disintegration times were lower than 5 min for all samples and friability less than 0.5% for all tablet formulations. The hardness of the reference product (25.5N) was lower compared to the other tablet samples (30-80.3N). All tablet formulations reached 75% release after 5 min of the dissolution test, but none of the compounding formulations reached the minimum 75% release after 45 min, probably due to inadequate excipient composition and amount. On average, excipient accounted for 46.3% of the capsule weight (against 74% in tablets), and some of the products did not contain water-soluble substances to promote dissolution.Conclusion: The results of this study indicate a lack of quality in compounding nimesulide products, which could jeopardize patients' health and treatment.Â

Biodiversity of β-carboline profile of banisteriopsis caapi and ayahuasca, a plant and a brew with neuropharmacological potential

Ayahuasca is a psychoactive infusion with a large pharmacological application normally prepared with Banisteriopsis caapi, which contains the monoamine oxidase inhibitors β-carbolines, and Psichotria virids, which contains the serotonin receptor agonist N,N dimethyltryptamine (DMT). The objectives of this study were to investigate the chemical profile of B. caapi and of ayahuasca collected in various Brazilian regions. In total, 176 plant lianas, of which 159 B. caapi and 33 ayahuasca samples were analyzed. Dried liana samples were powdered, extracted with methanol, diluted, and analyzed by LC-MS/MS. Ayahuasca samples were diluted and analyzed. Mean concentrations in B. caapi were 4.79 mg/g harmine, 0.451 mg/g harmaline, and 2.18 mg/g tetrahydroharmine (THH), with a high variability among the samples (RSD from 78.9 to 170%). Native B. caapi samples showed significantly higher harmine concentrations than cultivated ones, and samples from the Federal District/Goiás had higher THH content than those collected in the State of Acre. The other Malpighiaceae samples did not contain β-carbolines, except for one D. pubipetala sample. Concentrations in ayahuasca samples ranged from 0.109 to 7.11 mg/mL harmine, 0.012 to 0.945 mg/mL harmaline, 0.09 to 3.05 mg/mL THH, and 0.10 to 3.12 mg/mL DMT. The analysis of paired ayahuasca/B. caapi confirmed that harmine is reduced to harmaline and to THH during the brew preparation. This is the largest study conducted with Malpighiaceae samples and showed a large variability in the main β-carbolines present in B. caapi. This biodiversity is a challenge for standardization of the material used in ethnopharmacological studies of B. caapi and ayahuasca

Photography-based taxonomy is inadequate, unnecessary, and potentially harmful for biological sciences

The question whether taxonomic descriptions naming new animal species without type specimen(s) deposited in collections should be accepted for publication by scientific journals and allowed by the Code has already been discussed in Zootaxa (Dubois & Nemésio 2007; Donegan 2008, 2009; Nemésio 2009a–b; Dubois 2009; Gentile & Snell 2009; Minelli 2009; Cianferoni & Bartolozzi 2016; Amorim et al. 2016). This question was again raised in a letter supported by 35 signatories published in the journal Nature (Pape et al. 2016) on 15 September 2016. On 25 September 2016, the following rebuttal (strictly limited to 300 words as per the editorial rules of Nature) was submitted to Nature, which on 18 October 2016 refused to publish it. As we think this problem is a very important one for zoological taxonomy, this text is published here exactly as submitted to Nature, followed by the list of the 493 taxonomists and collection-based researchers who signed it in the short time span from 20 September to 6 October 2016

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Perfil químico da ayahuasca e da espécie Banisteriopsis caapi e seu potencial anti-inflamatório em cultura de células BV-2

Tese (doutorado)—Universidade de Brasília, Faculdade de Saúde, Programa de Pós-Graduação em Ciências Farmacêuticas, 2021.A ayahuasca é uma bebida psicotrópica com grande potencial terapêutico preparada principalmente pela decocção do cipó de Banisteriopsis caapi e das folhas da Psychotria viridis. Este estudo teve como objetivos caracterizar o perfil químico de indivíduos de B. caapi e de amostras de ayahuasca coletadas em diversas regiões do país e avaliar a atividade biológica do extrato de B. caapi e seus componentes em células de micróglia BV-2. No total, 33 amostras de ayahuasca e 176 amostras de cipó da família Malpighiaceae foram coletadas, sendo 159 B. caapi. Os cipós secos foram triturados e extraídos com metanol e a ayahuasca diluída para análise por LC-MS/MS. As concentrações médias das amostras de B. caapi foram de 4,79 mg/g de harmina, 0,451 mg/g de harmalina e 2,18 mg/g de THH, com uma alta variabilidade entre as amostras (DPR de 78,9 a 170%). Amostras nativas de B. caapi tiveram concentração de harmina maior do que as cultivadas e amostras do DF/GO tiveram maior concentração de THH do que as amostras do Acre. Concentrações nas amostras de ayahuasca variaram de 0,109 a 7,11 mg/mL de harmina, 0,012 a 0,945 mg/mL de harmina, 0,09 a 3,05 mg/mL de THH e 0,10 a 3,12 mg/mL de DMT. Foi confirmada a hipótese de que a harmina é reduzida a harmalina e então a THH durante a decocção no preparo da ayahuasca. Uma amostra de B. caapi do tipo ourinho coletada em Goiás foi submetida a cromatografia semipreparativa (HPLC-DAD) para isolamento de potenciais novos compostos. As frações isoladas foram analisadas por LC-MS/MS do tipo TOF de alta resolução para determinação de massa exata: F1 – 174,0918 e 233,1289; F2 – 353,1722; F3 – 304,3001; F4 – 188,1081; F5 – 205,0785. A atividade biológica das frações, do extrato de B. caapi e das ß-carbolinas harmina, harmalina e THH foi avaliada por meio dos testes de viabilidade celular, apoptose/necrose, produção de EROS e produção de citocinas por células microgliais BV-2, cultivadas em DMEM. A maioria dos tratamentos resultou em ação anti inflamatória com redução na produção de citocinas pró-inflamatórias, em especial as frações F4 e F5 e as ß-carbolinas. Os resultados sugerem que compostos presentes no extrato de B. caapi têm potencial para tratar doenças neurodegenerativas, uma vez que o excesso de ativação microglial pró-inflamatória está envolvido na neuropatologia de doenças do SNC.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) e Fundação de Apoio a Pesquisa do Distrito Federal (FAP/DF).Ayahuasca is psychoactive brew with important therapeutic potential that is prepared by the decoction of Banisteriopsis caapi vine and Psychotria viridis leaves. The main objectives of this study were to investigate the chemical profile of B. caapi and of ayahuasca collected in various Brazilian regions and to evaluate the biological activity of B. caapi extract and its components in BV-2 microglial cells. In total, 33 ayahuasca samples and 176 Malpighiaceae lianas, of which 159 B. caapi, were collected. Dried liana samples were powdered, extracted with methanol, diluted, and analyzed by LC-MS/MS. Ayahuasca samples were diluted and analyzed. Mean concentrations in B. caapi were 4.79 mg/g harmine, 0.451 mg/g harmaline, and 2.18 mg/g tetrahydroharmine (THH), with a high variability among the samples (RSD from 78.9 to 170%). Native B. caapi samples showed significantly higher harmine concentrations than cultivated ones, and samples from the Federal District/Goiás had higher THH content than those collected in the State of Acre. Concentrations in ayahuasca samples ranged from 0.109 to 7.11 mg/mL harmine, 0.012 to 0.945 mg/mL harmaline, 0.09 to 3.05 mg/mL THH, and 0.10 to 3.12 mg/mL DMT. The analysis of paired ayahuasca/B. caapi confirmed that harmine is reduced to harmaline and to THH during the brew preparation. A B. caapi sample of “ourinho” type was submitted to semipreparative chromatography (HPLC-DAD) for the isolation of potentially new compounds. Isolated samples were analyzed by high resolution LC-MS/MS TOF for exact mass determination: F1 – 174.0918 e 233.1289; F2 – 353.1722; F3 – 304.3001; F4 – 188.1081; F5 – 205.0785. Biological activity of fractions, B. caapi extract and ß-carbolines harmine, harmaline and THH was evaluated by cellular viability and apoptosis/necrosis assays and ROS and cytokines production by BV-2 microglial cells. Most of the treatments resulted in anti-inflammatory effects with the reduction of proinflammatory cytokines production, especially fractions F4 and F5 and the ß-carbolines. The results suggest that the compounds present in B. caapi extract have the potential to treat neurodegenerative diseases, as the excess of proinflammatory microglial activation is involved in the neuropathology of CNS diseases

Components of Banisteriopsis caapi, a Plant Used in the Preparation of the Psychoactive Ayahuasca, Induce Anti-Inflammatory Effects in Microglial Cells

Banisteriopsis caapi is used to prepare the psychoactive beverage ayahuasca, and both have therapeutic potential for the treatment of many central nervous system (CNS) conditions. This study aimed to isolate new bioactive compounds from B. caapi extract and evaluate their biological activity, and that of the known &beta;-carboline components of the plant (harmine, harmaline, and tetrahydroharmine), in BV-2 microglial cells, the in vivo activation of which is implicated in the physiopathology of CNS disorders. B. caapi extract was fractionated using semipreparative liquid chromatography (HPLC-DAD) and the exact masses ([M + H]+m/z) of the compounds in the 5 isolated fractions were determined by high-resolution LC-MS/MS: F1 (174.0918 and 233.1289), F2 (353.1722), F3 (304.3001), F4 (188.1081), and F5 (205.0785). Harmine (75.5&ndash;302 &micro;M) significantly decreased cell viability after 2 h of treatment and increased the number of necrotic cells and production of reactive oxygen species at equal or lower concentrations after 24 h. F4 did not impact viability but was also cytotoxic after 24 h. Most treatments reduced proinflammatory cytokine production (IL-2, IL-6, IL-17, and/or TNF), especially harmaline and F5 at 2.5 &micro;M and higher concentrations, tetrahydroharmine (9.3 &micro;M and higher), and F5 (10.7 &micro;M and higher). The results suggest that the compounds found in B. caapi extract have anti-inflammatory potential that could be explored for the development of treatments for neurodegenerative diseases

Components of Banisteriopsis caapi, a plant used in the preparation of the psychoactive ayahuasca, induce anti-inflammatory effects in microglial cells

Banisteriopsis caapi is used to prepare the psychoactive beverage ayahuasca, and both have therapeutic potential for the treatment of many central nervous system (CNS) conditions. This study aimed to isolate new bioactive compounds from B. caapi extract and evaluate their biological activity, and that of the known β-carboline components of the plant (harmine, harmaline, and tetrahydroharmine), in BV-2 microglial cells, the in vivo activation of which is implicated in the physiopathology of CNS disorders. B. caapi extract was fractionated using semipreparative liquid chromatography (HPLC-DAD) and the exact masses ([M + H]+m/z) of the compounds in the 5 isolated fractions were determined by high-resolution LC-MS/MS: F1 (174.0918 and 233.1289), F2 (353.1722), F3 (304.3001), F4 (188.1081), and F5 (205.0785). Harmine (75.5–302 µM) significantly decreased cell viability after 2 h of treatment and increased the number of necrotic cells and production of reactive oxygen species at equal or lower concentrations after 24 h. F4 did not impact viability but was also cytotoxic after 24 h. Most treatments reduced proinflammatory cytokine production (IL-2, IL-6, IL-17, and/or TNF), especially harmaline and F5 at 2.5 µM and higher concentrations, tetrahydroharmine (9.3 µM and higher), and F5 (10.7 µM and higher). The results suggest that the compounds found in B. caapi extract have anti-inflammatory potential that could be explored for the development of treatments for neurodegenerative diseases