Publicação
Caracterização da afinidade da interação de fármacos com albumina do soro, comparação de várias metodologias
| Resumo: | “Anyone who wishes to have more than an extremely superficial understanding of life in all its various manifestations need biochemistry” – Hans Krebs. The study of the interactions between drugs and protein is of great interest and value, the equilibrium and dynamics between molecule and macromolecule determines the physico-chemical properties that the system will have under the conditions in which it is. It is, therefore, of utmost importance to understand the mechanisms that rule these interactions and their consequences in the system. Therefore, the objective of the present work is to investigate the interaction of drugs with serum albumin, to estimate their association constants and dynamics of interaction, so that the system described here serves as an in vitro model for the parameters of these interactions. Chapter 1, was based on Chapters 3 and 4 of Lehningher's Principles of Biochemistry, and describes the amino acids, proteins and their structures, as well as aspects that justify the formation of peptide bonds, and structural conformations adopted by peptides and proteins, in primary, secondary and tertiary structures. The forces that lead to the formation of these complexes and justify their structuring. In Chapter 2 was presented the binding of molecules to plasma proteins and their implications in the properties of the drugs in plasma, the bound fraction and their consequences on the drug metabolism, half-life, volume of distribution and clearance. In Chapter 3 were described the techniques used to estimate the Ka association constants, through the hydrogels of BSA-PEG, the thermodynamic parameters of the interactions Ka, ΔG, ΔH and TΔS, through Isothermic Titration Calorimetry (ITC), and in the end were presented the drugs selected for the study (salicylic acid, diclofenac, chlorpromazine, acetaminophen, verapamil and labetalol) and the reasons for being chosen. Chapter 4 contains a description of the experimental conditions used in the tests with their respective methods, characterization of the activated polyethylene glycol and reagents used in these tests. In chapter 5 are the results obtained and all their analysis, from where the association constants for all drugs were obtained with the use of the hydrogels of BSA-PEG in accordance with the literature, thermodynamic parameters in the ITC for salicylic acid and diclofenac. Finally, in Chapter 6 is summarized all the work done in the conclusions drawn, the advantages and disadvantages of each technique as well as the application of the methodologies to optimize an in vitro model that is able to elucidate and predict the in vivo properties of the active ingredients used in medicine. |
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| Autores principais: | Paiva, Victor Santos de |
| Assunto: | Plasma Protein Binding Equilibrium and Dynamics BSA-PEG Hydrogels Isothermal Titration Calorimetry Ligação às Proteínas do Plasma Equilíbrio e Dinâmica Hidrogéis BSA-PEG Calorimetria de Titulação Isotérmica |
| Ano: | 2023 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade de Coimbra |
| Idioma: | português |
| Origem: | Estudo Geral - Universidade de Coimbra |
| Resumo: | “Anyone who wishes to have more than an extremely superficial understanding of life in all its various manifestations need biochemistry” – Hans Krebs. The study of the interactions between drugs and protein is of great interest and value, the equilibrium and dynamics between molecule and macromolecule determines the physico-chemical properties that the system will have under the conditions in which it is. It is, therefore, of utmost importance to understand the mechanisms that rule these interactions and their consequences in the system. Therefore, the objective of the present work is to investigate the interaction of drugs with serum albumin, to estimate their association constants and dynamics of interaction, so that the system described here serves as an in vitro model for the parameters of these interactions. Chapter 1, was based on Chapters 3 and 4 of Lehningher's Principles of Biochemistry, and describes the amino acids, proteins and their structures, as well as aspects that justify the formation of peptide bonds, and structural conformations adopted by peptides and proteins, in primary, secondary and tertiary structures. The forces that lead to the formation of these complexes and justify their structuring. In Chapter 2 was presented the binding of molecules to plasma proteins and their implications in the properties of the drugs in plasma, the bound fraction and their consequences on the drug metabolism, half-life, volume of distribution and clearance. In Chapter 3 were described the techniques used to estimate the Ka association constants, through the hydrogels of BSA-PEG, the thermodynamic parameters of the interactions Ka, ΔG, ΔH and TΔS, through Isothermic Titration Calorimetry (ITC), and in the end were presented the drugs selected for the study (salicylic acid, diclofenac, chlorpromazine, acetaminophen, verapamil and labetalol) and the reasons for being chosen. Chapter 4 contains a description of the experimental conditions used in the tests with their respective methods, characterization of the activated polyethylene glycol and reagents used in these tests. In chapter 5 are the results obtained and all their analysis, from where the association constants for all drugs were obtained with the use of the hydrogels of BSA-PEG in accordance with the literature, thermodynamic parameters in the ITC for salicylic acid and diclofenac. Finally, in Chapter 6 is summarized all the work done in the conclusions drawn, the advantages and disadvantages of each technique as well as the application of the methodologies to optimize an in vitro model that is able to elucidate and predict the in vivo properties of the active ingredients used in medicine. |
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