A carregar...
Publicação
Aplicação dos princípios do Quality by Design ao processo de um antibiótico
| Nome: | Descrição: | Tamanho: | Formato: | |
|---|---|---|---|---|
| 601.66 KB | Adobe PDF |
Autores
Orientador(es)
Resumo(s)
Atualmente, os medicamentos são obrigatoriamente produzidos por unidades industriais altamente
qualificadas e autorizadas pelas entidades reguladoras, cabendo a ambos a garantia da aplicação de
padrões de qualidade exigentes. Assim, a implementação do Quality by Design (QbD) tem vindo a
aumentar, pois avalia a qualidade de um produto ao longo do seu processo.
No âmbito desta dissertação, aplicou-se os princípios do QbD ao processo de formação de um
antibiótico, o composto 1, que surge numa mistura com 3 compostos/impurezas (2-4), biossintetisados
pelo microrganismo Pseudomonas fluorescens. Surgem ainda outras impurezas (7-13), relevantes no
processo de fermentação. O processo de obtenção de (1), foi obtido através de transferência de
tecnologia (TT) e passa pela sua extração a partir de um permeado obtido por fermentação, sendo a fase
orgânica resultante, descolorada com carvão ativado e precipitada a uma concentração de 200 g/L, de
forma a obter um pó final rico em (1) e com uma percentagem de impurezas inferior a 6 %.
Para implementar a metodologia do QbD ao processo de (1), identificaram-se os parâmetros críticos
do processo, através dos ensaios realizados aos vários passos do processo (pH, temperatura, extração,
tratamento com carvão e agente secante, precipitação, filtração e secagem), onde se verificou que o pH,
o rácio de solvente de extração: permeado, a lavagem do filtro de dicalite (terra de diatomáceas), a
concentração da fase orgânica antes da precipitação e a lavagem do pó final, são críticos neste processo.
Para avaliar o risco associado a cada uma destas operações realizou-se a avaliação do risco, onde foi
possível diminuir o risco elevado associado à concentração, através da adição de um ponto de controlo
antes da precipitação.
A fim de confirmar os doseamentos de (1) obtidos por HPLC, técnica utilizada como método de
rotina para o doseamento de (1) e respetivas impurezas ao longo dos ensaios realizados, realizou-se a
validação de um método de quantificação por 1H qRMN, com recurso a um composto de pureza
conhecida, produzido na Cipan. Após a pré-validação desta metodologia de RMN, avaliou-se a sua
aplicabilidade a outros antibióticos sintetizados na Cipan, sendo que os resultados obtidos se encontram
em concordância com resultados anteriores. Por fim, para a realização da quantificação de (1), realizou se a sua caracterização completa através de RMN com recurso a experiências 1D e 2D, seguida da
respetiva quantificação, que se demonstrou em concordância com os doseamentos obtidos por HPLC.
Portanto, concluiu-se que esta metodologia foi validada para (1) e pode ser também aplicada em
compostos estruturalmente muito distintos.
Currently, active pharmaceutical ingredients must be produced by highly qualified industrial units authorized by the regulatory authorities, both of which are responsible for ensuring that demanding quality standards are applied. Thus, the implementation of Quality by Design (QbD) has been increasing, as it assesses the quality of a product throughout its process. Within the scope of this dissertation, the principles of QbD were applied to the formation process of compound (1), an antibiotic, which appears in a mixture with 3 compounds/impurities (2-4), biosynthesized by the microorganism Pseudomonas fluorescens. Other impurities (7-13), relevant to the fermentation process, also appear. The extraction process of (1) was obtained through technology transfer (TT) and begins with the extration of (1) from a permeate obtained through fermentation, where the resulting organic phase is treated with activated charcoal and precipitated to 200 g/L, to obtain a powder rich in (1) and with an impurity percentage of less than 6 %. To implement the QbD methodology in the process of (1), the critical parameters of the process were identified through the tests carried out to different stages in the process (pH, temperature, extraction ratio and solvents, use of drying agent and activated charcoal, precipitation, filtration and drying), where we gathered that, pH, extraction solvent:permeate ratio, dicalite (diatomaceous earth) filter wash, organic phase concentration and the washing of the final powder, were critical in this process. Furthermore, the risk associated with each of the critical parameters was accessed, where with corrective measure, we were able to reduce the risk associated with the organic phase concentration, by adding a control point before precipitation. To confirm the assay of (1) obtained by HPLC, a technique used as a routine method for the assay of (1) and its associated impurities throughout the tests that were carried out, a 1H qNMR quantification was validated using a compound of known purity, produced at Cipan. After the pre-validation of the NMR methodology, its applicability was evaluated with other antibiotics produced at Cipan, and the results obtained were in agreement with previous results. Lastly, in order to quantify compound 1, its complete characterization was carried out using 1D and 2D NMR experiments, followed by its quantification, which appeared to be in agreement with the assays obtained via HPLC. It was therefore concluded that this methodology was validated for the quantification of (1), as well as compounds with different structures.
Currently, active pharmaceutical ingredients must be produced by highly qualified industrial units authorized by the regulatory authorities, both of which are responsible for ensuring that demanding quality standards are applied. Thus, the implementation of Quality by Design (QbD) has been increasing, as it assesses the quality of a product throughout its process. Within the scope of this dissertation, the principles of QbD were applied to the formation process of compound (1), an antibiotic, which appears in a mixture with 3 compounds/impurities (2-4), biosynthesized by the microorganism Pseudomonas fluorescens. Other impurities (7-13), relevant to the fermentation process, also appear. The extraction process of (1) was obtained through technology transfer (TT) and begins with the extration of (1) from a permeate obtained through fermentation, where the resulting organic phase is treated with activated charcoal and precipitated to 200 g/L, to obtain a powder rich in (1) and with an impurity percentage of less than 6 %. To implement the QbD methodology in the process of (1), the critical parameters of the process were identified through the tests carried out to different stages in the process (pH, temperature, extraction ratio and solvents, use of drying agent and activated charcoal, precipitation, filtration and drying), where we gathered that, pH, extraction solvent:permeate ratio, dicalite (diatomaceous earth) filter wash, organic phase concentration and the washing of the final powder, were critical in this process. Furthermore, the risk associated with each of the critical parameters was accessed, where with corrective measure, we were able to reduce the risk associated with the organic phase concentration, by adding a control point before precipitation. To confirm the assay of (1) obtained by HPLC, a technique used as a routine method for the assay of (1) and its associated impurities throughout the tests that were carried out, a 1H qNMR quantification was validated using a compound of known purity, produced at Cipan. After the pre-validation of the NMR methodology, its applicability was evaluated with other antibiotics produced at Cipan, and the results obtained were in agreement with previous results. Lastly, in order to quantify compound 1, its complete characterization was carried out using 1D and 2D NMR experiments, followed by its quantification, which appeared to be in agreement with the assays obtained via HPLC. It was therefore concluded that this methodology was validated for the quantification of (1), as well as compounds with different structures.
Descrição
Tese de Mestrado, Química, 2024, Universidade de Lisboa, Faculdade de Ciências
Palavras-chave
Pseudomonas fluorescens Quality by Design (QbD) Metodologias de Extração e Recristalização Quantificação 1H qRMN Teses de mestrado - 2024