Pharmacokinetics of neutron-irradiated meglumine antimoniate in Leishmania amazonensis-infected BALB/c mice

Cutaneous leishmaniasis (CL) is a parasitic disease caused by the protozoan Leishmania spp. Pentavalent antimonial agents have been used as an effective therapy, despite their side effects and resistant cases. Their pharmacokinetics remain largely unexplored. This study aimed to investigate the pharmacokinetic profile of meglumine antimoniate in a murine model of cutaneous leishmaniasis using a radiotracer approach. Methods: Meglumine antimoniate was neutron-irradiated inside a nuclear reactor and was administered once intraperitoneally to uninfected and L. amazonensis-infected BALB/c mice. Different organs and tissues were collected and the total antimony was measured. Results: Higher antimony levels were found in infected than uninfected footpad (0.29% IA vs. 0.14% IA, p = 0.0057) and maintained the concentration. The animals accumulated and retained antimony in the liver, which cleared slowly. The kidney and intestinal uptake data support the hypothesis that antimony has two elimination pathways, first through renal excretion, followed by biliary excretion. Both processes demonstrated a biphasic elimination profile classified as fast and slow. In the blood, antimony followed a biexponential open model. Infected mice showed a lower maximum concentration (6.2% IA/mL vs. 11.8% IA/mL, p = 0.0001), a 2.5-fold smaller area under the curve, a 2.7-fold reduction in the mean residence time, and a 2.5-fold higher clearance rate when compared to the uninfected mice. Conclusions: neutron-irradiated meglumine antimoniate concentrates in infected footpad, while the infection affects antimony pharmacokinetics.(AU)

Antimonial drugs entrapped into phosphatidylserine liposomes: physicochemical evaluation and antileishmanial activity

Abstract: INTRODUCTION: Leishmaniasis is a disease caused by the protozoan Leishmania that resides mainly in mononuclear phagocytic system tissues. Pentavalent antimonials are the main treatment option, although these drugs have toxic side effects and high resistance rates. A potentially alternative and more effective therapeutic strategy is to use liposomes as carriers of the antileishmanial agents. The aims of this study were to develop antimonial drugs entrapped into phosphatidylserine liposomes and to analyze their biological and physicochemical characteristics. METHODS: Liposomes containing meglumine antimoniate (MA) or pentavalent antimony salt (Sb) were obtained through filter extrusion (FEL) and characterized by transmission electron microscopy. Promastigotes of Leishmania infantum were incubated with the drugs and the viability was determined with a tetrazolium dye (MTT assay). The effects of these drugs against intracellular amastigotes were also evaluated by optical microscopy, and mammalian cytotoxicity was determined by an MTT assay. RESULTS: Liposomes had an average diameter of 162nm. MA-FEL showed inhibitory activity against intracellular L. infantum amastigotes, with a 50% inhibitory concentration (IC50) of 0.9μg/mL, whereas that of MA was 60μg/mL. Sb-FEL showed an IC50 value of 0.2μg/mL, whereas that of free Sb was 9μg/mL. MA-FEL and Sb-FEL had strong in vitro activity that was 63-fold and 39-fold more effective than their respective free drugs. MA-FEL tested at a ten-times higher concentration than Sb-FEL did not show cytotoxicity to mammalian cells, resulting in a higher selectivity index. CONCLUSIONS: Antimonial drug-containing liposomes are more effective against Leishmania-infected macrophages than the non-liposomal drugs.

O SUS na medicina nuclear do Brasil: avaliação e comparação dos dados fornecidos pelo Datasus e CNEN / SUS in nuclear medicine in Brazil: analysis and comparison of data provided by Datasus and CNEN

Objetivo: Investigar o acesso a procedimentos ambulatoriais de medicina nuclear por intermédio do Sistema Único de Saúde (SUS) do Brasil e analisar a correspondência dos dados fornecidos por este sistema com os da Comissão Nacional de Energia Nuclear (CNEN). Materiais e Métodos: Foram obtidos e avaliados os dados disponíveis no Datasus quanto a quantidade de câmaras de cintilação, procedimentos ambulatoriais de 2008 a 2012, esfera administrativa responsável por estes procedimentos, tipo de prestador de serviços e terceirização de serviços. Também foi feita comparação com os dados de estabelecimentos autorizados pela CNEN. Resultados: O estudo mostrou que ainda falta amadurecimento do sistema quanto à sua completa alimentação, especialmente no campo de equipamentos disponíveis. Foi possível elencar os procedimentos mais realizados e verificar o crescimento da especialidade no período estudado. Estabelecimentos privados são responsáveis pela maior parte dos procedimentos cobertos pelo SUS. Entretanto, muitos estabelecimentos de saúde não são autorizados pela CNEN. Conclusão: O Datasus oferece dados importantes para uma análise como a feita neste estudo, embora alguns pontos ainda demandem atenção. O trabalho mostrou, quantitativamente, a realidade brasileira quanto ao acesso a procedimentos de medicina nuclear oferecidos pelo/para o SUS. .

Objective: To investigate the outpatient access to nuclear medicine procedures by means of the Brazilian Unified Health System (SUS), analyzing the correspondence between data provided by this system and those from Comissão Nacional de Energia Nuclear (CNEN) (National Commission of Nuclear Energy). Materials and Methods: Data provided by Datasus regarding number of scintillation chambers, outpatient procedures performed from 2008 to 2012, administrative responsibility for such procedures, type of service providers and outsourced services were retrieved and evaluated. Also, such data were compared with those from institutions certified by CNEN. Results: The present study demonstrated that the system still lacks maturity in terms of correct data input, particularly regarding equipment available. It was possible to list the most common procedures and check the growth of the specialty along the study period. Private centers are responsible for most of the procedures covered and reimbursed by SUS. However, many healthcare facilities are not certified by CNEN. Conclusion: Datasus provides relevant data for analysis as done in the present study, although some issues still require attention. The present study has quantitatively depicted the Brazilian reality regarding access to nuclear medicine procedures offered by/for SUS. .

Biodistribution of meglumine antimoniate in healthy and Leishmania (Leishmania) infantum chagasi-infected BALB/c mice

Pentavalent antimonials such as meglumine antimoniate (MA) are the primary treatments for leishmaniasis, a complex disease caused by protozoan parasites of the genus Leishmania . Despite over 70 years of clinical use, their mechanisms of action, toxicity and pharmacokinetics have not been fully elucidated. Radiotracer studies performed on animals have the potential to play a major role in pharmaceutical development. The aims of this study were to prepare an antimony radiotracer by neutron irradiation of MA and to determine the biodistribution of MA in healthy and Leishmania (Leishmania) infantum chagasi-infected mice. MA (Glucantime(r)) was neutron irradiated inside the IEA-R1 nuclear reactor, producing two radioisotopes, 122Sb and 124Sb, with high radionuclidic purity and good specific activity. This irradiated compound presented anti-leishmanial activity similar to that of non-irradiated MA in both in vitro and in vivo evaluations. In the biodistribution studies, healthy mice showed higher uptake of antimony in the liver than infected mice and elimination occurred primarily through biliary excretion, with a small proportion of the drug excreted by the kidneys. The serum kinetic curve was bi-exponential, with two compartments: the central compartment and another compartment associated with drug excretion. Radiotracers, which can be easily produced by neutron irradiation, were demonstrated to be an interesting tool for answering several questions regarding antimonial pharmacokinetics and chemotherapy.

Tc-99m direct radiolabeling of monoclonal antibody ior egf/r3: quality control and image studies in mice

Monoclonal antibodies (Mabs) have been useful for immunoscintigraphic applications in clinical diagnosis since they were introduced in the practice of nuclear medicine. The ior egf/r3 (Centis, Cuba) is a murine monoclonal antibody against epidermal growth factor receptor (EGF-R) and has been widely used in the radioimmunodiagnosis of tumors of epithelial origin. Labeled with 99mTc, its main application in Nuclear Medicine is the follow up, detection and evaluation of tumor recurrences. The objective of this work is to describe the preparation of a lyophilized formulation (kit) for radiolabeling the Mab ior egf/r3 with 99mTc for immunoscintigraphic applications. Radiolabeling efficiency, effects on immunoreactivity, image studies and stability of the formulation are reported. The study demonstrated that the kit formulation can be labeled with 99mTc at high yields and can be used to visualize in vivo human tumors of epithelial origin by immunoscintigraphy studies.

Anticorpos monoclonais (AcM) tem sido usado para aplicações imunocintilográficas no diagnóstico clínico desde sua introdução na prática da medicina nuclear. O ior egf/r3 (Centis, Cuba) é um anticorpo monoclonal murino contra o receptor do fator do crescimento epidérmico (EGF-R) e tem sido usado extensivamente no radioimunodiagnóstico de tumores de origem epitelial. Marcado com 99mTc sua maior aplicação em Medicina Nuclear é: acompanhamento, detecção e avaliação de recorrências tumorais. O objetivo deste trabalho é descrever a preparação da formulação liofilizada (kit) para radiomarcação do AcM ior egf/r3 com 99mTc para aplicações imunocintilográficas. A eficiência da radiomarcação, efeito sobre a imunorreatividade, estudos de imagem e estabilidade da formulação são relatados. O estudo demonstrou que a formulação do kit pode ser marcada com 99mTc com alto rendimento e pode ser usado para visualizar in vivo tumores humanos de origem epitelial por estudos imunocintilográficos.

Study of gels of molybdenum with cerium in the preparation of generators of 99Mo - 99mTc

99mTc has ideal nuclear properties for organ imaging in nuclear medicine, and it is obtained from the 99Mo-99mTc generator. Four different types of generators are available: chromatographic that uses 99Mo from fission of uranium; MEK solvent extraction; Tc2O7 sublimation; gel chromatographic. This work presents the preparation of gel generators of molybdenum with cerium and characterization of the gels: mass ratio between molybdenum and cerium, structure, size of particles and elution percentage of 99mTc after irradiating the gels. Eight gels were prepared at the same temperature of 50 °C with concentrations of NaOH of 2 and 4 mol/L, mass ratio of 0.31 and 0.38 and final pH of 3.5 and 4.5. The analysis of the results proved that these gels are not adequate for preparation of the generators of 99Mo-99mTc, since the elution percentages are low, when compared with the gel of molybdenum with zirconium.

O 99mTc é o radiofármaco mais utilizado em Medicina Nuclear. Ele é obtido do gerador de 99Mo-99mTc e existem quatro tipos diferentes de geradores: cromatográfico que utiliza 99Mo de fissão; extração por solvente com MKT; sublimação do heptaóxido de tecnécio; cromatográfico tipo gel. Este trabalho apresenta a preparação de geradores tipo gel de molibdênio com cério, a caracterização desses géis com relação à quantidade de molibdênio e de cério, sua estrutura, tamanho das partículas e porcentagem de eluição do 99mTc após o gel ser irradiado. Foram preparados oito géis na temperatura de 50°C com concentração de NaOH de 2 e 4 mol/L, relação de massa de 0,31 e 0,38 e pH final de 3,5 e 4,5. A análise dos resultados comprovou que esses géis não são adequados para preparação dos geradores de 99Mo-99mTc, já que as porcentagens de eluição são baixas, quando comparadas com o gel de molibdênio com zircônio.

In Vitro antileishmanial properties of neutron-irradiated meglumine antimoniate

Pentavalent antimony, as meglumine antimoniate (Glucantime® ) or sodium stibogluconate (Pentostam® ), is the main treatment for leishmaniasis, a complex of diseases caused by the protozoan Leishmania, and an endemic and neglected threat in Brazil. Despite over half a century of clinical use, their mechanism of action, toxicity and pharmacokinetic data remain unknown. The analytical methods for determination of antimony in biological systems remain complex and have low sensitivity. Radiotracer studies have a potential in pharmaceutical development. The aim of this study was to obtain a radiotracer for antimony, with suitable physical and biological properties. Meglumine antimoniate was neutron irradiated inside the IEA-R1 nuclear reactor, producing two radioisotopes 122Sb and 124Sb, with high radionuclidic purity and good specific activity. This compound showed the same antileishmanial activity as the native compound. The use of the radiotracers, easily created by neutron irradiation, could be an interesting tool to solve important questions in antimonial pharmacology.

Os antimoniais pentavalentes, como o antimoniato de meglumina (Glucantime® ) ou estibogluconato de sódio (Pentostam® ), são o principal tratamento para a leishmaniose, um complexo de doenças causadas pelo protozoário parasita Leishmania, uma doença endêmica e negligenciada no Brasil. Apesar do seu uso clínico por mais de meio século, seu mecanismo de ação, toxicidade e dados de farmacocinética permanecem desconhecidos. Os métodos analíticos para determinação de antimônio em sistemas biológicos são complexos e apresentam baixa sensibilidade. Estudos utilizando radiotraçadores têm papel potencial no desenvolvimento farmacológico. O objetivo deste estudo foi desenvolver um radiotraçador de antimônio, com propriedades físicas e biológicas adequadas. O antimoniato de meglumina foi irradiado por nêutrons no reator nuclear IEA-R1, produzindo dois radioisótopos: 122Sb e 124Sb, com alta pureza radionuclídica e boa atividade específica. Este composto mostrou atividade antileishmania similar ao fármaco não irradiado. O uso de radiotraçadores, facilmente produzidos por irradiação por nêutrons pode ser um importante instrumento para elucidar questões sobre a farmacologia dos antimoniais.

Liquid kit for preparation of 188rhenium-etidronate

The aim of this study was the preparation of a liquid kit for radiolabeling of 188Re-HEDP (hydroxyethylidene diphosphonate). 188Re was obtained from alumina based 188W/188Re generators. This paper reports the efficacy of a cold kit stored for more than two weeks, determined by the dependence of the radiolabeling yields of 188Re-HEDP on the incubation time, reducing agent concentration, the effects of concentration of ligand, the pH of the reaction and the temperature. The cold kits showed a good stability when carrie-free rhenium-188 was added in the reaction mixture.

O objetivo desse trabalho é a preparação de um kit líquido para a radiomarcação do 188Re-HEDP (hidroxietilideno - difosfonato). O 188Re foi obtido por meio do gerador 188W/188Re em alumina. Este trabalho reporta a experiência do preparo de um kit frio armazenado por até duas semanas. Os parâmetros estudados para obter melhores rendimentos de marcação do 188Re-HEDP foram: o tempo de incubação, a concentração do agente redutor, massa do ligante, o pH e a temperatura da reação. O kit frio mostrou uma boa establidade quando o 188Re livre de carregador foi adicionado na mistura de reação.