Preparation, characterization and evaluation of the in vivo trypanocidal activity of ursolic acid-loaded solid dispersion with poloxamer 407 and sodium caprate

Ursolic acid is a promising candidate for treatment of Chagas disease; however it has low aqueous solubility and intestinal absorption, which are both limiting factors for bioavailability. Among the strategies to enhance the solubility and dissolution of lipophilic drugs, solid dispersions are growing in popularity. In this study, we employed a mixture of the surfactants poloxamer 407 with sodium caprate to produce a solid dispersion containing ursolic acid aimed at enhancing both drug dissolution and in vivo trypanocidal activity. Compared to the physical mixture, the solid dispersion presented higher bulk density and smaller particle size. Fourier Transform Infrared Spectroscopy results showed hydrogen bonding intermolecular interactions between drug and poloxamer 407. X-ray diffractometry experiments revealed the conversion of the drug from its crystalline form to a more soluble amorphous structure. Consequently, the solubility of ursolic acid in the solid dispersion was increased and the drug dissolved in a fast and complete manner. Taken together with the oral absorption-enhancing property of sodium caprate, these results explained the increase of the in vivo trypanocidal activity of ursolic acid in solid dispersion, which also proved to be safe by cytotoxicity evaluation using the LLC-MK2 cell line.O ácido ursólico é um candidato promissor para o tratamento da doença de Chagas, contudo este fármaco possui baixa solubilidade aquosa e limitada absorção intestinal, ambos os fatores limitantes da biodisponibilidade. Entre as estratégias para potencializar a solubilidade e a dissolução de fármacos lipofílicos, as dispersões sólidas estão crescendo em popularidade. Neste estudo, empregamos mistura dos tensoativos, poloxamer 407 e caprato de sódio, para produzir dispersão sólida contendo ácido ursólico, com o objetivo de aumentar tanto a dissolução do fármaco quanto a atividade tripanocida in vivo. Comparada à mistura física, a dispersão sólida apresentou maior densidade e menor tamanho de partícula. Os resultados da análise de espectroscopia no infravermelho com transformada de Fourier mostraram interações intermoleculares do tipo ligações de hidrogênio entre o fármaco e o poloxamer 407. Os experimentos de difratometria de raio-X revelaram a conversão do fármaco de sua forma cristalina para a forma amorfa, mais solúvel. Consequentemente, a solubilidade do ácido ursólico em dispersão sólida foi aumentada e o fármaco dissolveu-se de maneira mais rápida e completa. Em conjunto com as propriedades promotoras de absorção oral do caprato de sódio, estes resultados explicaram o aumento da atividade tripanocida in vivo do ácido ursólico em dispersão sólida, que também se provou segura após avaliação de citotoxicidade empregando a linhagem celular LLC-MK2

HPLC assay of lidocaine in in vitro dissolution test of the Poloxamer 407 gels

Apresenta-se método simples de cromatografia líquida de alta eficiência (CLAE) para análise da lidocaína em meio aquoso, após estudo de liberação in vitro. A lidocaína foi analisada usando-se coluna LichroCART RP-18 (5 mm, 125x4 mm), fase móvel constituída de acetonitrila: tampão fosfato de sódio 0,05 M, pH 6 (35:65), adicionada de 0,05% de dietilamina com fluxo de 1 mL/min. O tempo de retenção foi de 7,9 min. O comprimento de onda de análise utilizado foi de 210 nm. A linearidade do método foi de 1,25 a 25 µg/mL com coeficiente de variação intraensaio e inter-ensaio menor que 3,5 %. A metodologia desenvolvida e validada mostrou sensibilidade e especificidade para a realização dos estudos propostos, considerando-se que as amostras obtidas a partir dos estudos de liberação in vitro contêm concentrações muito baixas do fármaco, além de outras substâncias do meio de dissolução que podem interferir no doseamento. A quantificação do fármaco e dos interferentes pode não ser possível se for efetuada por outras metodologias analíticas convencionais. Assim, o método desenvolvido é de grande importância para a quantificação do fármaco nas alíquotas obtidas nos ensaios de liberação in vitro.A simple high performance liquid chromatography method to assay lidocaine hydrochloride in aqueous receiving media, following in vitro release, is presented. Lidocaine hydrochloride was analysed using a 5 mm LichroCART® RP-18 column (125 x 4 mm i.d.). The mobile phase was acetonitrile: 0.05 M sodium phosphate buffer, pH 6.0 (35:65) and 0.05% of diethylamine at a flow rate of 1 mL/min. The retention time was 7.9 min. Detection was carried out at 210 nm at room temperature (28 ºC). The method was found to be linear in the range 1.25 to 25 mg/mL, showing average intraassay and inter-assay coefficients of variation below 3.5%. The proposed method was validated for linearity, specificity, precision and accuracy and was shown to be useful for the analysis of lidocaine hydrochloride in in vitro release studies

Nanocarriers for Nitric Oxide Delivery

Nitric oxide (NO) is a promising pharmaceutical agent that has vasodilative, antibacterial, and tumoricidal effects. To study the complex and wide-ranging roles of NO and to facilitate its therapeutic use, a great number of synthetic compounds (e.g., nitrosothiols, nitrosohydroxyamines, N-diazeniumdiolates, and nitrosyl metal complexes) have been developed to chemically stabilize and release NO in a controlled manner. Although NO is currently being exploited in many biomedical applications, its use is limited by several factors, including a short half-life, instability during storage, and potential toxicity. Additionally, efficient methods of both localized and systemic in vivo delivery and dose control are needed. One strategy for addressing these limitations and thus increasing the utility of NO donors is based on nanotechnology

Solid Dispersion of Ursolic Acid in Gelucire 50/13: a Strategy to Enhance Drug Release and Trypanocidal Activity

Solid dispersions (SDs) are an approach to increasing the water solubility and bioavailability of lipophilic drugs such as ursolic acid (UA), a triterpenoid with trypanocidal activity. In this work, Gelucire 50/13, a surfactant compound with permeability-enhancing properties, and silicon dioxide, a drying adjuvant, were employed to produce SDs with UA. SDs and physical mixtures (PMs) in different drug/carrier ratios were characterized and compared using differential scanning calorimetry, hot stage microscopy, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), particle size, water solubility values, and dissolution profiles. Moreover, LLC-MK2 fibroblast cytotoxicity and trypanocidal activity evaluation were performed to determine the potential of SD as a strategy to improve UA efficacy against Chagas disease. The results demonstrated the conversion of UA from the crystalline to the amorphous state through XRD. FTIR experiments provided evidence of intermolecular interactions among the drug and carriers through carbonyl peak broadening in the SDs. These findings helped explain the enhancement of water solubility from 75.98 mu g/mL in PMs to 293.43 mu g/mL in SDs and the faster drug release into aqueous media compared with pure UA or PMs, which was maintained after 6 months at room temperature. Importantly, improved SD dissolution was accompanied by higher UA activity against trypomastigote forms of Trypanosoma cruzi, but not against mammalian fibroblasts, enhancing the potential of UA for Chagas disease treatment.Coordenacao de Aperfeicoamento de Pessoal de Nivel SuperiorConselho Nacional de Desenvolvimento Cientifico e Tecnologic

Estudo das características e propriedades da permeação in vitro de micropartículas de CMC/quitosana como sitema de liberação cutânea para vitamina E

Micropartículas de carboximetilcelulose (CMC)/quitosana contendo vitamina E foram preparadas pelo método de coacervação complexa e seu uso potencial como um sistema de liberação tópico foi avaliado. Estudos da morfologia, da distribuição do tamanho de partículas, da eficiência de encapsulação, da estabilidade física e química e da liberação e permeação cutânea in vitro foram realizados. As análises por Microscopia Eletrônica de Varredura mostraram que as partículas são esféricas, possuem uma superfície homogênea e ausência de agregados, com diâmetros na faixa de 2,7 a 7,6 µm. A eficiência de encapsulação da vitamina E foi 81%. Os estudos de estabilidade química mostraram proteção da vitamina E encapsulada, sendo que a diferença em relação à quantidade de ativo remanescente na emulsão O/A foi de 8,1% e na A/O, de 10,8%, após armazenamento a 45 °C por um período de 60 dias. O ensaio de liberação in vitro mostrou que 48% da vitamina E encapsulada, quantificada por CLAE, foram liberadas em 24 horas de experimento. Nos estudos de retenção e permeação in vitro observou-se que a emulsão A/O proporcionou maior penetração da vitamina E tanto da forma livre como encapsulada.Os sistemas avaliados parecem ser promissores para veiculação de ativos em preparações de uso tópico.Carboxymethylcellulose (CMC)/chitosan microparticles containing vitamin E were prepared by a complex coacervation method and their potential use as a topical delivery system was evaluated. Morphology, particle size distribution, encapsulation yield, physical and chemical stability, in vitro release and permeation through skin were studied. The microparticles appeared to be spherical, with a homogeneous surface and were not aggregated. Mean diameters ranged from 2.7 to 7.6 µm and the encapsulation yield was 81%. Chemical stability studies indicated a protection of encapsulated vitamin E, of 8.1% for O/W and of 10.83% for W/O emulsions, following storage at 45 °C for 60 days. Forty-eight% of vitamin E, determined by HPLC, were released within 24 hours. In vitro permeation and retention studies showed a higher penetration rate of vitamin E in the free and encapsulated forms, from the W/O emulsion. The carriers studied seem to be promising systems for topical administration

Propolis Standardized Extract (EPP-AF®), an Innovative Chemically and Biologically Reproducible Pharmaceutical Compound for Treating Wounds

The aim of this study was to develop a formulation, containing the propolis standardized extract (EPP-AF®), which can assist in the healing of skin lesions. To achieve this objective the antimicrobial activity and chemical composition of the propolis extract was determined. The final product was subjected to in vitro and in vivo pre-clinical evaluation. The broth macrodilution method was used to determine the antimicrobial activity of the extracts and formulations against the microorganisms most commonly found in burns, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus and Staphylococcus epidermidis. Wistar rats with puncture wounded skin were used to evaluate the wound healing properties of propolis. The results of chemical and biological characterization demonstrated the batch-to-batch reproducibility of the standardized extract which is an unprecedented result. The antimicrobial and wound healing activity of the pharmaceutical studied showed the best results when samples contain 3.6% propolis, suggesting that this is the most promising composition

Development and validation of a HPLC method for quantification of ursolic acid in solid dispersions

Ursolic acid is a natural molecule that presents several pharmacological properties. In this work, an analytical method by RP-HPLC has been developed and validated for quantification of this drug in the solid dispersions, using PEG 6000 and Poloxamer 407 as polymers. The method was specific, linear in the range of 1.0-50.0 µg mL-1 (r<0.99), precise (CV < 5% for both inter- and intra-assays), accurate (maximum deviation of ± 13%), and robust to the parameters evaluated. This method has proved to be simple and useful for ursolic acid determination in solid dispersions, enabling its determination in pharmaceutical dosage form

Description of the complexity of prescribed medication regimens in primary health care of Ribeirão Preto - SP

Introduction: Pharmacotherapy is the main therapeutic resource for the management of diseases. However, the number of drugs prescribed, dose frequency, and mode of administration can make the treatment more complex and influence treatment outcomes. The aim of this study was to measure the complexity of prescribed medication regimens in primary health care (PHC) services in Ribeirão Preto, Brazil. Methods: This cross-sectional study included 1,009 participants: 889 from primary health units and 120 from family health units in Ribeirão Preto, Brazil. Treatment complexity was assessed using the Medication Regimen Complexity Index (MRCI). Results: MRCI mean scores were 12.5 points (SD = 9.3) and dose frequency was the major contributor to increase the score. The complexity of pharmacotherapy showed a significant correlation with the number of prescribed medications (r = 0.93, p < 0.01), but not with patients' age (r = 0.28, p < 0.01). There is also no difference in complexity between the sexes (p = 0.83) and the types of primary health care service (p = 0.31). An analysis of variance revealed that patients with lower levels of education receive more complex prescriptions (p < 0.01). Conclusions: The pharmacotherapy prescribed in PHC services from Ribeirão Preto, Brazil is complex, and there is a need to concentrate efforts and adopt strategies to simplify drug prescription without compromising patient's clinical status. Keywords: Primary health care; drug prescriptions; drug therapy; medication regimen complexit

Description of the complexity of prescribed medication regimens in primary health care of Ribeirão Preto - SP

Introduction: Pharmacotherapy is the main therapeutic resource for the management of diseases. However, the number of drugs prescribed, dose frequency, and mode of administration can make the treatment more complex and influence treatment outcomes. The aim of this study was to measure the complexity of prescribed medication regimens in primary health care (PHC) services in Ribeirão Preto, Brazil. Methods: This cross-sectional study included 1,009 participants: 889 from primary health units and 120 from family health units in Ribeirão Preto, Brazil. Treatment complexity was assessed using the Medication Regimen Complexity Index (MRCI). Results: MRCI mean scores were 12.5 points (SD = 9.3) and dose frequency was the major contributor to increase the score. The complexity of pharmacotherapy showed a significant correlation with the number of prescribed medications (r = 0.93, p < 0.01), but not with patients' age (r = 0.28, p < 0.01). There is also no difference in complexity between the sexes (p = 0.83) and the types of primary health care service (p = 0.31). An analysis of variance revealed that patients with lower levels of education receive more complex prescriptions (p < 0.01). Conclusions: The pharmacotherapy prescribed in PHC services from Ribeirão Preto, Brazil is complex, and there is a need to concentrate efforts and adopt strategies to simplify drug prescription without compromising patient's clinical status. Keywords: Primary health care; drug prescriptions; drug therapy; medication regimen complexit

Determinação de heparina fracionada em preparações farmacêuticas utilizando turbidimetria

A turbidimetric method has been used for quantification of fractionated heparin (FH) in pharmaceutical dosage. The UV detection at two wavelengths (290 and 500 nm) showed a significant increase in sensitivity of the method, specificity, and linearity to range 5.0-50.0 µg mL-1 and 50.0-200.0 µg mL-1, respectively (r < 0.99). At both wavelengths, the method was precise (inter-assay CV < 5.0%, and intra-assay CV < 3.0%), accurate (maximum deviation of ± 12%), and robust to the parameters evaluated. Turbidimetry proved to be easy, inexpensive and relatively fast. The results obtained attest to the reliability of the method