Estudos de dimerização de peptídeos e os efeitos na inibição da atividade de DNA topoisomerases
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Data
2021-06-16
Autores
Angelo, Jonatas Medeiros de Almeida
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Universidade Estadual Paulista (Unesp)
Resumo
As doenças infeciosas estão entre as principais causas de mortalidade em todo o mundo, especialmente devido à crescente disseminação de bactérias patogênicas resistentes aos antimicrobianos atualmente disponíveis. Diante deste cenário, a pesquisa e o desenvolvimento de novos antimicrobianos, que possam atuar em diferentes alvos intracelulares, torna-se de grande urgência e importância. DNA topoisomerases são enzimas essenciais para a regulação dos processos fisiológicos e viabilidade celular e têm se mostrado um alvo bem atrativo para a ação de vários fármacos. Neste sentido, a pesquisa de novos inibidores de topoisomerases, incluindo os peptídeos sintéticos, representa uma estratégia promissora no desenvolvimento de uma nova classe de agentes terapêuticos. O uso de peptídeos como antimicrobianos apresenta importantes vantagens, como alta especificidade, diversidade nos mecanismos de ação, relativa facilidade no processo de síntese, capacidade de impedir a formação de biofilme e raras ocorrências de resistência bacteriana. Assim sendo, a síntese de análogos dimerizados do peptídeo WRWYCRCK se mostra uma importante e eficiente abordagem, que pode vir a contribuir com o desenvolvimento de novos e mais potentes inibidores de topoisomerases, candidatos a agentes antimicrobianos. Neste trabalho, peptídeos lineares, análogos ao octapeptídeo WRWYCRCK, foram obtidos pela metodologia da fase sólida e submetidos a processos oxidativos para a formação de ligações dissulfeto direcionadas, com o objetivo de avaliar o efeito do número e posição destas ligações, na inibição da atividade de diferentes topoisomerases. A dimerização favoreceu a atividade inibitória do octapeptídeo WRWYCRCK sobre as DNA topoisomerases, independentemente da classe e da origem das mesmas. A maior susceptibilidade das topoisomerases aos dímeros DimLin2 e DimAPLin2 sugere que ambos os resíduos de cisteína e as respectivas ligações dissulfeto são fundamentais para a inibição enzimática, o que corrobora o efeito bactericida e fungicida obtidos a baixas concentrações para estes peptídeos. Os peptídeos sintetizados não apresentaram atividade hemolítica e a inibição do crescimento de células de fibroblastos da linhagem NIH/3T3 foi detectada apenas em concentrações superiores às necessárias para a inibição enzimática. De uma maneira geral, os resultados obtidos se mostraram bastante promissores, uma vez que a dimerização direcionada da sequência peptídica WRWYCRCK resultou na obtenção de novos inibidores de DNA topoisomerases, com grande potencial para o desenvolvimento de novos agentes antimicrobianos.
Infectious diseases are among the leading causes of mortality worldwide, especially due to the increasing spread of antimicrobial-resistant pathogenic bacteria currently available. Given of this scenario, the research and development of new antimicrobial drugs, which can act on different intracellular targets, become urgent and of utmost importance. DNA topoisomerases are enzymes essential for the physiological processes regulation and cell viability and have been shown a very attractive target for various drugs. In this context, the search for novel topoisomerase inhibitors, including synthetic peptides, represents a promising strategy in the development of a new class of therapeutic agents. The use of synthetic peptides as antimicrobial drugs has important advantages, such as high specificity, diversity in the mechanisms of action, relative ease in the synthesis process, ability to prevent the formation of biofilms and rare bacterial resistance occurrences. Therefore, the synthesis of dimerized analogues of the WRWYCRCK peptide proves to be an important and efficient approach, which may contribute to the development of new and more potent topoisomerase inhibitors, candidates for antimicrobial agents. In this work, linear peptides, analogous to the WRWYCRCK octapeptide, were obtained by the solid phase methodology and subjected to oxidative processes for the targeted disulfide bonds formation, in order to evaluate the effect of the number and position of these bonds, in the inhibition of the activity of different topoisomerases. Dimerization favored the inhibitory activity of the octapeptide WRWYCRCK on DNA topoisomerases, regardless of their class and origin. The greater susceptibility of topoisomerases to DimLin2 and DimAPLin2 dimers suggests that both cysteine residues and the respective disulfide bonds are fundamental for enzymatic inhibition, which corroborates the bactericidal and fungicidal effects obtained at low concentrations for these peptides. The synthesized peptides did not show hemolytic activity and the inhibition of the growth of fibroblast cells of the NIH/3T3 strain was detected only in concentrations higher than those required for enzymatic inhibition did. In general, the results obtained showed to be quite promising, since the targeted dimerization of the WRWYCRCK peptide sequence resulted in the obtaining of new DNA topoisomerase inhibitors, with great potential for the development of new antimicrobial agents.
Infectious diseases are among the leading causes of mortality worldwide, especially due to the increasing spread of antimicrobial-resistant pathogenic bacteria currently available. Given of this scenario, the research and development of new antimicrobial drugs, which can act on different intracellular targets, become urgent and of utmost importance. DNA topoisomerases are enzymes essential for the physiological processes regulation and cell viability and have been shown a very attractive target for various drugs. In this context, the search for novel topoisomerase inhibitors, including synthetic peptides, represents a promising strategy in the development of a new class of therapeutic agents. The use of synthetic peptides as antimicrobial drugs has important advantages, such as high specificity, diversity in the mechanisms of action, relative ease in the synthesis process, ability to prevent the formation of biofilms and rare bacterial resistance occurrences. Therefore, the synthesis of dimerized analogues of the WRWYCRCK peptide proves to be an important and efficient approach, which may contribute to the development of new and more potent topoisomerase inhibitors, candidates for antimicrobial agents. In this work, linear peptides, analogous to the WRWYCRCK octapeptide, were obtained by the solid phase methodology and subjected to oxidative processes for the targeted disulfide bonds formation, in order to evaluate the effect of the number and position of these bonds, in the inhibition of the activity of different topoisomerases. Dimerization favored the inhibitory activity of the octapeptide WRWYCRCK on DNA topoisomerases, regardless of their class and origin. The greater susceptibility of topoisomerases to DimLin2 and DimAPLin2 dimers suggests that both cysteine residues and the respective disulfide bonds are fundamental for enzymatic inhibition, which corroborates the bactericidal and fungicidal effects obtained at low concentrations for these peptides. The synthesized peptides did not show hemolytic activity and the inhibition of the growth of fibroblast cells of the NIH/3T3 strain was detected only in concentrations higher than those required for enzymatic inhibition did. In general, the results obtained showed to be quite promising, since the targeted dimerization of the WRWYCRCK peptide sequence resulted in the obtaining of new DNA topoisomerase inhibitors, with great potential for the development of new antimicrobial agents.
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Palavras-chave
Inibidores enzimáticos, Antibióticos peptídicos, Antibacterianos, Anti-infecciosos, Dímeros