AVALIAÇÃO DA EQUIVALÊNCIA FARMACÊUTICA EM COMPRIMIDOS DE HIDROCLOROTIAZIDA FABRICADOS POR COMPRESSÃO DIRETA E GRANULAÇÃO VIA ÚMIDA

Objetivos: Avaliar a equivalência farmacêutica dos comprimidos de hidroclotiaziada de 50 miligramas produzidos por via direta e por via úmida, fabricados em aula do curso de farmácia na Universidade do Extremo Sul Catarinense. Metodologia: Foi realizado teste para a avaliação da velocidade de dissolução dos comprimidos de hidroclorotiazida. O teste foi realizado seguindo parâmetros estabelecidos pela farmacopeia brasileira. Resultados: Submetemos os comprimidos testes ao aparelho de dissolutor, para o hidroclotiazida referência (Clorana) obtivemos valores de dissolução iguais a 96,4% em 30 minutos, para comprimidos produzidos em aula por via direta, valores iguais a 71,2% em 30 minutos e produzidos por via úmida, valores iguais a 110,9% em 30 minutos. Como exigido pela farmacopeia, os valores para a dissolução são acima de 60% da substância dissolvida em 30 minutos. Como preconizado na monografia da substância, os valores da concentração do analíto deve ser 93% a 107% da quantidade declarada. Neste quesito, só o referência obteve resultado satisfatório com valor de 101,9% da substância, quando por via direta teve valor de 75,6% e via úmida em 115,5% da substância declarada. Conclusão: O processo de fabricação de um fármaco pode causar uma alteração na sua disponibilidade, como testado in vitro pelo teste de dissolução. Para os testes observamos que não alcançamos os parâmetros exigidos pela monografia da substância.Palavras-chaves: Equivalência Farmacêutica; Dissolução; Hidroclorotiazida

AVALIAÇÃO DA EQUIVALÊNCIA FARMACÊUTICA EM COMPRIMIDOS DE HIDROCLOROTIAZIDA FABRICADOS POR COMPRESSÃO DIRETA E GRANULAÇÃO VIA ÚMIDA

Objetivos: Avaliar a equivalência farmacêutica dos comprimidos de hidroclotiaziada de 50 miligramas produzidos por via direta e por via úmida, fabricados em aula do curso de farmácia na Universidade do Extremo Sul Catarinense. Metodologia: Foi realizado teste para a avaliação da velocidade de dissolução dos comprimidos de hidroclorotiazida. O teste foi realizado seguindo parâmetros estabelecidos pela farmacopeia brasileira. Resultados: Submetemos os comprimidos testes ao aparelho de dissolutor, para o hidroclotiazida referência (Clorana) obtivemos valores de dissolução iguais a 96,4% em 30 minutos, para comprimidos produzidos em aula por via direta, valores iguais a 71,2% em 30 minutos e produzidos por via úmida, valores iguais a 110,9% em 30 minutos. Como exigido pela farmacopeia, os valores para a dissolução são acima de 60% da substância dissolvida em 30 minutos. Como preconizado na monografia da substância, os valores da concentração do analíto deve ser 93% a 107% da quantidade declarada. Neste quesito, só o referência obteve resultado satisfatório com valor de 101,9% da substância, quando por via direta teve valor de 75,6% e via úmida em 115,5% da substância declarada. Conclusão: O processo de fabricação de um fármaco pode causar uma alteração na sua disponibilidade, como testado in vitro pelo teste de dissolução. Para os testes observamos que não alcançamos os parâmetros exigidos pela monografia da substância.Palavras-chaves: Equivalência Farmacêutica; Dissolução; Hidroclorotiazida

Immunodiagnosis of Canine Visceral Leishmaniasis Using Mimotope Peptides Selected from Phage Displayed Combinatorial Libraries

ELISA and RIFI are currently used for serodiagnosis of canine visceral leishmaniasis (CVL). The accuracy of these tests is controversial in endemic areas where canine infections by Trypanosoma cruzi may occur. We evaluated the usefulness of synthetic peptides that were selected through phage display technique in the serodiagnosis of CVL. Peptides were chosen based on their ability to bind to IgGs purified from infected dogs pooled sera. We selected three phage clones that reacted only with those IgGs. Peptides were synthesized, polymerized with glutaraldehyde, and used as antigens in ELISA assays. Each individual peptide or a mix of them was reactive with infected dogs serum. The assay was highly sensitive and specific when compared to soluble Leishmania antigen that showed cross-reactivity with anti-T. cruzi IgGs. Our results demonstrate that phage display technique is useful for selection of peptides that may represent valuable synthetic antigens for an improved serodiagnosis of CVL

PEPOP 2.0: new approaches to mimic non-continuous epitopes

International audienceBACKGROUND:Bioinformatics methods are helpful to identify new molecules for diagnostic or therapeutic applications. For example, the use of peptides capable of mimicking binding sites has several benefits in replacing a protein which is difficult to produce, or toxic. Using peptides is less expensive. Peptides are easier to manipulate, and can be used as drugs. Continuous epitopes predicted by bioinformatics tools are commonly used and these sequential epitopes are used as is in further experiments. Numerous discontinuous epitope predictors have been developed but only two bioinformatics tools have been proposed so far to predict peptide sequences: Superficial and PEPOP 2.0. PEPOP 2.0 can generate series of peptide sequences that can replace continuous or discontinuous epitopes in their interaction with their cognate antibody

PEPOP: new approaches to mimic non-continous epitopes

Bioinformatics methods are helpful to identify new molecules for diagnostic or therapeutic applications. For example, the use of peptides capable of mimicking binding sites has several benefits as replacing a protein difficult to produce, or toxic. Using peptides is less expensive. Peptides are easier to manipulate, and can be used as drugs. Continuous epitope predicted by bioinformatics tools are commonly used and these sequential epitopes are used as such in further experiments. Numerous discontinuous epitope predictors have been developed but only two bioinformatics tools proposed so far to predict peptide sequences: Superficial and PEPOP. PEPOP can generate series of peptide sequences that can replace continuous or discontinuous epitopes in their interaction with their cognate antibody. We have developed an improved version of PEPOP dedicated to answer to the experimentalists' need for a tool able to handle proteins and to turn them into peptides. The PEPOP web site has been reorganized by peptide prediction category and is therefore better formulated to experimental designs. Since the first version of PEPOP, 32 new methods of peptide design were developed. In total, PEPOP proposes 35 methods in which 34 deal specifically with discontinuous epitopes, the most represented epitope type in nature. We present the user-friendly, well-structured web-site of PEPOP and its validation through the use of predicted immunogenic or antigenic peptides mimicking discontinuous epitopes in different experimental ways. PEPOP proposes 35 methods of peptide design to guide experimentalists in using peptides potentially capable of replacing the cognate protein in its interaction with an Ab

Use of a Synthetic Biosensor for Neutralizing Activity-Biased Selection of Monoclonal Antibodies against Atroxlysin-I, an Hemorrhagic Metalloproteinase from Bothrops atrox Snake Venom

International audienc

Genome-wide screening and identification of new Trypanosoma cruzi antigens with potential application for chronic Chagas disease diagnosis.

The protozoan Trypanosoma cruzi is the etiologic agent of Chagas disease, an infection that afflicts approximately 8 million people in Latin America. Diagnosis of chronic Chagas disease is currently based on serological tests because this condition is usually characterized by high anti-T. cruzi IgG titers and low parasitemia. The antigens used in these assays may have low specificity due to cross reactivity with antigens from related parasite infections, such as leishmaniasis, and low sensitivity caused by the high polymorphism among T. cruzi strains. Therefore, the identification of new T. cruzi-specific antigens that are conserved among the various parasite discrete typing units (DTUs) is still required. In the present study, we have explored the hybrid nature of the T. cruzi CL Brener strain using a broad genome screening approach to select new T. cruzi antigens that are conserved among the different parasite DTUs and that are absent in other trypanosomatid species. Peptide arrays containing the conserved antigens with the highest epitope prediction scores were synthesized, and the reactivity of the peptides were tested by immunoblot using sera from C57BL/6 mice chronically infected with T. cruzi strains from the TcI, TcII or TcVI DTU. The two T. cruzi proteins that contained the most promising peptides were expressed as recombinant proteins and tested in ELISA experiments with sera from chagasic patients with distinct clinical manifestations: those infected with T. cruzi from different DTUs and those with cutaneous or visceral leishmaniasis. These proteins, named rTc_11623.20 and rTc_N_10421.310, exhibited 94.83 and 89.66% sensitivity, 98.2 and 94.6% specificity, respectively, and a pool of these 2 proteins exhibited 96.55% sensitivity and 98.18% specificity. This work led to the identification of two new antigens with great potential application in the diagnosis of chronic Chagas disease

Neutralization of Atr-I or <i>B. atrox</i> venom enzymatic activities on Abz-LVEALYQ-EDDnp substrate.

<p>Purified mAbatrs were pre-incubated with Atr-I or <i>B. atrox</i> venom at 37°C for 30 minutes previously to addition of the FRET substrate. Results are normalized to Atr-I or <i>B. atrox</i> venom alone (positive control) and represent means ±S.D. of triplicates.</p

Cross-reactivity of mAbatrs with different toxins from <i>B. atrox</i> venom analyzed by western blotting.

<p><i>B. atrox</i> crude venom was transferred to a nitrocellulose membrane and incubated with rabbit polyclonal anti-Atr-I serum (<b>C</b>) as control, or mAbatr1 (<b>1</b>), mAbatr2 (<b>2</b>), mAbatr3 (<b>3</b>) or mAbatr6 (<b>6</b>). All mAbatrs recognized bands around 55, 30, 23 and 15 kDa.</p