Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Andrea Krejzová Supervisor: PharmDr. Ivan Vokřál, Ph.D. Title of diploma thesis: Effect of albendazole on the activity of selected enzymes in tapeworm Hymenolepis diminuta The efficacy of anthelmintics used to treat diseases caused by helminths is not always sufficient, and in some cases, we are directly facing resistance to these drugs. Helminths, including tapeworms, are able to defend against the toxic effect of anthelmintics using several mechanisms. Xenobiotic metabolizing enzymes and transport proteins belong to these mechanisms. When xenobiotic metabolizing enzymes are induced, the efficacy of therapy may be significantly reduced. The effect of xenobiotic metabolizing enzymes on the drug resistance development has been already described in number of helminths. In tapeworms this information is still missing. Main aim of this study was to determine effect of drug albendazole on the activity of selected xenobiotic metabolizing enzymes in rat tapeworm (Hymenolepis diminuta). Tapeworms were incubated with albendazole (1 μM and 10 μM) for 24 hours. Then activities of selected enzymes in cytosol-like, microsome-like and mitochondria-like fractions were determined. This study is focused on...

As parasitoses causadas por helmintos constituem um grave problema sanitário, tanto para os seres humanos quanto para os animais, além de gerar grandes prejuízos econômicos. O sulfóxido de albendazol é um fármaco anti-helmíntico de amplo espectro, largamente utilizado na medicina veterinária, veiculado pelas vias oral e parenteral, mediante a utilização de formas farmacêuticas convencionais. Apresenta biodisponibilidade baixa e irregular em função de sua pouca solubilidade nos fluidos biológicos. Para a manutenção da concentração plasmática e completa eliminação dos parasitos, são necessárias administrações reiteradas, ocasionando transtornos decorrentes do manejo freqüente dos animais e do aumento do custo da terapia. O presente trabalho teve como objetivo desenvolver e caracterizar um sistema microparticulado para liberação sustentada de sulfóxido de albendazol, de modo que este pudesse permanecer no organismo dos animais pelo tempo suficiente para a completa eliminação dos parasitos após uma única administração. Os sistemas foram obtidos a utilizando as técnicas de spray-drying e emulsificação / evaporação de solvente, tendo sido utilizados os polímeros Eudragit RS 30 D® e Eudragit RS PO®, respectivamente. As micropartículas obtidas foram caracterizadas com relação ao tamanho, à morfologia e à eficiência de encapsulação. Através da técnica de emulsificação / evaporação de solvente, foram obtidas partículas com diâmetro médio inferior a 300nm, estreita faixa de distribuição de tamanho e eficiência de encapsulação de aproximadamente 60%. Os resultados do estudo in vitro do perfil de liberação do fármaco a partir das micropartículas obtidas mostraram que, apesar de o sistema desenvolvido não ter sido capaz de sustentar a liberação do fármaco, o mesmo promoveu um aumento significativo da solubilidade do sulfóxido de albendazol em pH 7,4, fato este que pode contribuir para o aumento da biodisponibilidade do mesmo após administração parenteral. / The helminthosis are a serious sanitary problem, as for the men than for the animals, besides the great economic lacks. Albendazole sulphoxide is an antihelminthic drug with broad spectrum of action, widely used at veterinarian medicine, throw oral and parentereal vies, in conventional pharmaceutical dosages. It has low and irregular bioavailability due its low solubility in the biological fluids. For the maintenance of the plasmatic concentration and complete elimination of the parasites, it is required several administrations, creating many troubles due the frequent handling of the animals and increase in the costs of the therapy. The present work had as objective to develop and characterize microparticles for sustained release of albendazole sulphoxide, in order that the drug could be for a longer time in the animals organisms and the parasites could be eliminated after just one administration. The referred microparticles were obtained from the spray-drying and emulsification / evaporation of solvent techniques, using the polymers Eudragit RS 30 D® and Eudragit RS PO®, respectively. The obtained systems were characterized considering size, morphology and encapsulation efficiency. Using the emulsification / evaporation of solvent technique, it was prepared microparticles with medium diameter under 300nm, narrow range of size distribution and encapsulation efficiency of about 60%. The results of the in vitro release profile study of the drug from the prepared microparticles showed that besides the developed system was not be able to sustain the drug delivery, it was able to improve significantly the solubility of albendazole sulphoxide at pH 7.4, what can be useful to improve its parenteral bioavailability.

Half a billion school aged children suffer from anemia, with the majority of anemia caused by iron deficiency. Researchers have shown a strong correlation between low hemoglobin levels and presence of intestinal parasites in children with anemia. Childhood anemia has profound negative effects on physical growth, maturation, and cognitive development leading to poorer educational achievement. Using hemoglobin as a measure of anemia, this quasi-experimental study investigated impact of either iron supplementation or an antiparasitic medication on hemoglobin levels in two groups of children in a rural region of Eastern Ghana. Surprisingly, after a 6-month intervention period, hemoglobin levels in both groups significantly decreased. Further research is needed toinvestigate other factors impacting nutrition and incidence of anemia in pediatric populations in developing countries.

No description available.

Submitted by MARCEL KENZO VILALBA ONIZUKA null (marcel.kenzo@gmail.com) on 2016-11-22T17:07:43Z No. of bitstreams: 1 Dissertação_Marcel_Kenzo_Vilalba_Onizuka.pdf: 966239 bytes, checksum: 3186b3d5035f1c543aa30c82f00aad7c (MD5) / Approved for entry into archive by Felipe Augusto Arakaki (arakaki@reitoria.unesp.br) on 2016-11-25T15:49:39Z (GMT) No. of bitstreams: 1 onizuka_mkv_me_jabo.pdf: 966239 bytes, checksum: 3186b3d5035f1c543aa30c82f00aad7c (MD5) / Made available in DSpace on 2016-11-25T15:49:39Z (GMT). No. of bitstreams: 1 onizuka_mkv_me_jabo.pdf: 966239 bytes, checksum: 3186b3d5035f1c543aa30c82f00aad7c (MD5) Previous issue date: 2016-10-20 / O objetivo deste trabalho foi avaliar o desenvolvimento ponderal de bovinos precoces, tratados estrategicamente contra nematódeos gastrintestinais, em julho e setembro, com dois fármacos distintos, o sulfóxido de albendazole 10% e ivermectina 1%. O experimento teve duração de 112 dias e foi dividido em três etapas: E1 (pasto); E2 (pasto e suplemento); E3 (confinamento e eutanásia). Foram utilizados 36 bovinos (Braford), idade entre 18 e 20 meses, peso médio inicial 357 kg e naturalmente infectados com nematódeos gastrintestinais. Os animais foram distribuídos em três grupos experimentais (n=12), de acordo com peso e OPG: GI (controle); GII (sulfóxido de albendazole 10%); GIII (ivermectina 1%). Para análise dos dados foram avaliadas as variáveis peso, ganho em peso, OPG, helmintos remanescentes e rendimento de carcaça. Ao término do período experimental os animais foram eutanasiados para realizar a necropsia parasitológica. Em todas as análises estabeleceu-se como nível de significância 5%. Houve diferença significativa nas variáveis peso, ganho em peso, OPG e rendimento de carcaça quando comparou-se o GII aos demais grupos. Na necropsia parasitológica foi encontrada predominância das espécies Trichostrongylus axei e Cooperia punctata. Concluiu-se que os bovinos tratados estrategicamente com sulfóxido de albendazole 10%, em julho e setembro, apresentaram melhor desenvolvimento ponderal em relação aos demais grupos. / The objective of this study was to evaluate the weight development of early cattle strategically treated against gastrointestinal nematodes in July and September, with two different drugs, albendazole sulphoxide 10% and ivermectin 1%. The experiment lasted 112 days and was divided into three stages: S1 (pasture); S2 (pasture and supplement); S3 (feedlot and euthanasia). Were used 36 cattle (Braford), aged between 18 and 20 months, average weight 357 kg and naturally infected with gastrointestinal nematodes. The animals were divided into three experimental groups (n= 12), according to weight and EPG: GI (control); GII (albendazole sulphoxide 10%); GIII (ivermectin 1%). For data analysis was evaluated the variables weight, weight gain, EPG, remaining helminths and carcass yield. At the end of the experimental period the animals were euthanized to perform the parasitological necropsy. In all analyzes it was established as a significance level of 5%. There were significant differences in the variables weight, weight gain, EPG and carcass yield when compared to the GII to other groups. In parasitological necropsy found predominance of species Trichostrongylus axei and Cooperia punctata. It was concluded that cattle strategically treated with albendazole sulphoxide 10%, in July and September, showed better growth development than the other groups.

Orientador: Alvimar José da Costa / Resumo: O objetivo deste trabalho foi avaliar o desenvolvimento ponderal de bovinos precoces, tratados estrategicamente contra nematódeos gastrintestinais, em julho e setembro, com dois fármacos distintos, o sulfóxido de albendazole 10% e ivermectina 1%. O experimento teve duração de 112 dias e foi dividido em três etapas: E1 (pasto); E2 (pasto e suplemento); E3 (confinamento e eutanásia). Foram utilizados 36 bovinos (Braford), idade entre 18 e 20 meses, peso médio inicial 357 kg e naturalmente infectados com nematódeos gastrintestinais. Os animais foram distribuídos em três grupos experimentais (n=12), de acordo com peso e OPG: GI (controle); GII (sulfóxido de albendazole 10%); GIII (ivermectina 1%). Para análise dos dados foram avaliadas as variáveis peso, ganho em peso, OPG, helmintos remanescentes e rendimento de carcaça. Ao término do período experimental os animais foram eutanasiados para realizar a necropsia parasitológica. Em todas as análises estabeleceu-se como nível de significância 5%. Houve diferença significativa nas variáveis peso, ganho em peso, OPG e rendimento de carcaça quando comparou-se o GII aos demais grupos. Na necropsia parasitológica foi encontrada predominância das espécies Trichostrongylus axei e Cooperia punctata. Concluiu-se que os bovinos tratados estrategicamente com sulfóxido de albendazole 10%, em julho e setembro, apresentaram melhor desenvolvimento ponderal em relação aos demais grupos. / Mestre

A current initiative to eliminate lymphatic filariasis (LF), headed by the World Health Organization, aims to interrupt transmission of the disease through yearly community-wide treatment with the broad spectrum anthelmintic albendazole (ABZ), in combination with ivermectin (IVM) or diethylcarbamazine (DEC). Over the years, the use of both ABZ and IVM in the treatment of veterinary parasites has led to widespread anthelmintic resistance against these drugs. In this study, we genotyped microfilaria of Wuchereria bancrofti, a causative agent of LF, in order to detect the presence of mutations which confer ABZ resistance in other parasites, and we identified such mutations in worms obtained from untreated patients in Ghana and Burkina Faso, West Africa. Microfilaria from patients who had been treated with ABZ + IVM, had a significantly higher frequency of the resistant genotype, and this frequency was even higher in worms from patients that had received two rounds of treatment. In addition, the untreated population of microfilaria had an excess of homozygotes in the population. This excess homozygosity was equivalent to a Wright's Inbreeding Statistic of FIT= 0.44, and we found that the population was significantly subdivided between patients. In order to better understand the mechanisms and factors involved in the potential spread of ABZ resistance, caused by such mutations, through a population of Culex-transmitted W. bancrofti, we developed a deterministic model that incorporates genotype structure into the epidemiological model EPIFIL. This model predicts that the combination of ABZ + DEC leads to stronger selection for the resistant genotype than ABZ + IVM, and that drug efficacy assumptions are an important factor affecting the spread of drug resistance. Treatment coverage, non-random mating, initial allele frequency and number of treatments also had substantial impact on the speed and magnitude of the spread of ABZ resistance. When we expanded this model to include potential IVM-resistance alleles we found that, under ABZ + IVM treatment, selection for resistance to either drug is enhanced by the presence of resistance against the second drug. Similarly, excess homozygosity caused by parasite non-random mating may increase selection for a dominant IVM resistance allele through enhancing the spread of a recessive ABZ resistance allele. Resistence developed more slowly when it was inherited as a polygenic trait. Results from this study suggest that resistance monitoring is crucial, as resistance may not be apparent until treatment is stopped, recrudescence occurs and treatment is reapplied.

No description available.

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / O praziquantel (PZQ), um fÃrmaco quiral disponÃvel como racemato, e o albendazol (ABZ), um fÃrmaco biotransformado ao metabÃlito ativo quiral sulfÃxido de abendazol (ASOX), tem sido empregados no tratamento da neurocisticercose humana. O estudo abrange a investigaÃÃo da disposiÃÃo cinÃtica, metabolismo e enantiosseletividade na associaÃÃo ABZ - PZQ em voluntÃrios sadios. O estudo cruzado e aleatÃrio foi desenvolvido em trÃs fases (n=9), sendo que alguns voluntÃrios iniciaram pela FASE 1 (400mg de ABZ), outros pela FASE 2 (1500mg de PZQ) e outros pela FASE 3 (400mg de ABZ + 1500mg de PZQ). O perÃodo de washout foi de no mÃnimo 15 dias (FASE 1 seguida da FASE 2 e FASE 1 seguida da FASE 3) ou 7 dias (FASE 2 seguida de uma das outras FASES). As amostras seriadas de sangue foram coletadas no perÃodo de 0-48h. Os metabÃlitos do ABZ foram analisados por HPLC com detecÃÃo por fluorescÃncia e os enantiÃmeros do PZQ e do trans-4-hidroxipraziquantel (4-OHPZQ) foram analisados por LC-MS-MS. Os parÃmetros farmacocinÃticos foram determinados com auxÃlio do programa WinNonlin. O teste de Wilcoxon (p≤0.05) foi empregado para avaliar as razÃes enantiomÃricas de concentraÃÃes plasmÃticas do ASOX, PZQ e 4-OHPZQ. Os dados estÃo expressos como medianas. A disposiÃÃo cinÃtica do PZQ, 4-OHPZQ e do ASOX Ã enantiosseletiva na situaÃÃo de monoterapia; as razÃes de AUC sÃo de 2,97 para (+)-(S)-PZQ /(-)-(R)-PZQ, 0,78 para (+)-(S)-4OHPZQ /(-)-(R)-4-OHPZQ e 7,08 para (+)-ASOX/(-)-ASOX. A administraÃÃo de PZQ resulta em aumento das concentraÃÃes plasmÃticas do (+)-ASOX em 264% (AUC 980,42 vs 2591,80 ng.h/ml), do (-)-ASOX em 358% (139,59 vs 500,28 ng.h./ml) e do sulfona de albendazol em 187% (170,85 vs 319,50ng.h./ml) sugerindo o PZQ como inibidor da Pgp intestinal. A administraÃÃo de ABZ nÃo altera a disposiÃÃo cinÃtica do (+)-(S)-PZQ e dos metabÃlitos (-)-(R)-4-OHPZQ e (+)-(S)-4OHPZQ, mas resulta em aumento das concentraÃÃes plasmÃticas do (-)-(R)-PZQ em 64,77% (AUC 518,02 vs 853,57ng.h/ml) sugerindo inibiÃÃo enantiosseletiva do metabolismo do ASOX. Os dados permitem sugerir a possibilidade de aumento da eficÃcia terapÃutica na interaÃÃo ABZ-PZQ, embora outros estudos sejam necessÃrios para avaliar a seguranÃa da interaÃÃo. / The praziquantel (PZQ), a chiral drug available as racemic, and the albendazole (ABZ), a drug biotransformed into active metabolic chiral suphoxide of abendazol (ASOX), have been used in the treatment of human neurocysticercosis. The study covers the examination / search of the kinetic disposition, the metabolism, and the enantioselectiveness in the ABZ-PZQ association in healthy volunteers. The crossed and random study was developed in three phases (n=9), in which some volunteers started by PHASE 1 (400 mg of ABZ), others by PHASE 2 (1500mg of PZQ), and others by PHASE 3 (400 mg of ABZ + 1500mg of PZQ). The period of washout was of a minimum of 15 days (PHASE 1 followed by PHASE 2 and PHASE 1 followed by PHASE 3) or of 7 days (PHASE 2 followed by one of the other Phases). The serial blood samples were collected in a period of 0-48 hours. The ABZ metabolics were analised by HPLC with detection by fluorescence and the PZQ enantiomers and the trans-4-hydroxypraziquantel (4-OHPZQ) were analised by LC-MS-MS. The pharmacokinetic patterns were determined with the help of the WinNonlin program. The test of Wilcoxon (p≤0.05) was used to evaluate the enantiomer ratios of plasma concentrations of ASOX, PZQ and 4-OHPZQ. The data are shown as medians. The kinetic disposition of the PZQ, 4-OHPZQ and ASOX is enantioselective in the monotherapy situation; the ratios of AUC are of 2.97 to (+)-(S)-PZQ / (-)-(R)-PZQ, 0.78 to (+)-(S)-4-OHPZQ / (-)-(R)-4-OHPZQ, and 7.08 to (+)-ASOX / (-)-PZQ. The administration of the PZQ results in the increase of the plasma concentrations of the (+)-ASOX in 264% (AUC 980.42 vs 2591.80ng.h./ml), of the (-)-ASOX in 358% (139.59 vs 500.28ng.h./ml), and of the sulphona of albendazole in 187% (170.85 vs 319.50ng.h./ml), suggesting the PZQ as an inhibiting factor of the intestinal Pgp. The administration of the ABZ does not change/ alter the kinetic disposition of the (+)-(S)-PZQ, and of the metabolic (-)-(R)-4-OHPZQ and (+)-(S)-4-OHPZQ, but it results in the increase of the plasma concentrations of the (-)-(R)-PZQ in 64.77% (AUC 518.02 vs 853.57ng.h./ml ), suggesting enantioselective inhibition of the metabolism of the ASOX. The data allow us to suggest the possibility of increase of therapeutic efficacy in the ABZ-PZQ interaction; although, other studies are necessary to evaluate the safety of the interaction.

The aim of this research was to determine the influence of iron deficiency on both the efficacy and metabolic patterns of albendazole in mice infected with Heligmosomoides polygyrus. Anthelmintic efficacy was markedly decreased in iron-deficient mice; the deficiency was also associated with a decrease in body weight, altered hematological parameters and a decreased net egg output; worm establishment in the deficient group was not affected by the deficiency. Although anthelmintic efficacy was significantly decreased by the iron deficiency, plasma concentration profiles of the main metabolites, albendazole sulphoxide and albendazole sulphone, were not changed by the deficiency. Levels of intestinal cytochrome P-450, the main metabolizing enzyme of albendazole however, was significantly depressed in iron-deficient mice. These observations suggest that although pharmacokinetic parameters are not affected by iron deficiency, nutritional status has the potential to influence anthelmintic efficacy and thus warrants further study.