Estratégias analíticas integradas para obtenção de metabólitos citotóxicos a partir de Swinglea glutinosa e Penicillium setosum, um endofítico associado
Data
2021-10-08
Autores
Carvalho, Ana Calheiros de 
Orientadores
Veiga, Thiago André Moura
Tipo
Tese de doutorado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
Neste trabalho, foi conduzido o estudo químico da espécie vegetal Swinglea glutinosa (Rutaceae) e do metabolismo secundário de seus fungos endofíticos associados. Para isto, foi proposta a utilização da abordagem de desreplicação molecular via cromatografia líquida acoplada ao espectrômetro de massas de alta resolução (UHPLC-HRMS). O emprego dessa abordagem possibilitou, a partir de S. glutinosa, o isolamento de quatro substâncias não relacionadas ao gênero: lansamida I, lansiumamida B, N-(2-feniletil)cinamamida e lansiumamida C, as quais pertencem à classe das fenilacrilamidas. Além destas, também foram isolados cinco alcaloides acridônicos previamente conhecidos: citrusinina I, citibrasina, piranofolina, 5-hidroxinoracronicina e glicotricina IV. Paralelamente, foram obtidos 14 fungos endofíticos associados a S. glutinosa, os quais foram cultivados em quatro meios de cultura diferentes. Os micro-extratos obtidos foram analisados via HRMS e avaliados contra algumas linhagens de leucemia humana. Após a triagem inicial, decidiu-se priorizar o estudo químico de Penicillium setosum, um fungo recentemente descoberto. A partir desse microrganismo, foram isoladas seis substâncias pertencentes às classes das piranonas, dicetopiperazinas e antraquinonas cloradas, que foram identificadas como: 4-metil-5,6-dihidro-2H-piran-2-ona, felutanina C, bisdetiobis(metiltio)acetilaranotina, 7-cloroemodina, 2-cloro-1,3,8-trihidroxi-6-(hidroximetil)-antraquinona e ácido 2-cloroemódico. A felutanina C apresentou atividade contra a linhagem Kasumi-1, bem como seletividade para a mesma. Os experimentos realizados até o momento sugerem o mecanismo de morte por apoptose. Diante disso, este trabalho, além de enriquecer o conhecimento do perfil químico do gênero Swinglea e do fungo P. setosum, ofereceu excelentes perspectivas para a obtenção de substâncias com ação anti-leucemia.
In this work, the chemical study of Swinglea glutinosa (Rutaceae) plant species and the secondary metabolism of its associated endophytic fungi was carried out. For this, it was proposed to use the dereplication approach via liquid chromatography coupled to a high resolution mass spectrometer (UHPLC-HRMS). The use of dereplication methodologies allowed the isolation of four substances not reported for the Swinglea genus: lansamide I, lansiumamide B, N-(2-phenylethyl)cinnamamide and lansiumamide C, which belong to the class of phenylacrylamides. In addition to these, five previously known acridonic alkaloids were also isolated: citrusinine I, citibrasine, pyranofoline, 5-hydroxynorachronicine and glycothricine IV. In parallel, 14 endophytic fungi associated with S. glutinosa were obtained, which were cultivated in four different culture media. The micro-extracts obtained were dereplicated via HRMS and evaluated against human leukemia cell lines. After the initial screening, it was decided to prioritize the chemical study of Penicillium setosum, a recently discovered fungus. From this microorganism, six substances belonging to pyranones, diketopiperazines and chlorinated anthraquinones classes were isolated, identified as: 4- methyl-5,6-diihydro-2H-pyran-2-one, fellutanine C, bisdethiobis(methylthio)acetylaranotine, 7-chloroemodin, 2-chloro-1,3,8-trihydroxy-6- (hydroxymethyl)-anthraquinone and 2-chloroemodic acid. Fellutanin C showed promising activity against human leukemia cell line Kasumi-1 as well as selectivity for it. The experiments carried out so far indicate a mechanism of death by apoptosis. Therefore, this work, besides enriching the knowledge of the chemical profile of the genus Swinglea and the fungus P. setosum, offers great prospects for obtaining substances with anti-leukemia action.
In this work, the chemical study of Swinglea glutinosa (Rutaceae) plant species and the secondary metabolism of its associated endophytic fungi was carried out. For this, it was proposed to use the dereplication approach via liquid chromatography coupled to a high resolution mass spectrometer (UHPLC-HRMS). The use of dereplication methodologies allowed the isolation of four substances not reported for the Swinglea genus: lansamide I, lansiumamide B, N-(2-phenylethyl)cinnamamide and lansiumamide C, which belong to the class of phenylacrylamides. In addition to these, five previously known acridonic alkaloids were also isolated: citrusinine I, citibrasine, pyranofoline, 5-hydroxynorachronicine and glycothricine IV. In parallel, 14 endophytic fungi associated with S. glutinosa were obtained, which were cultivated in four different culture media. The micro-extracts obtained were dereplicated via HRMS and evaluated against human leukemia cell lines. After the initial screening, it was decided to prioritize the chemical study of Penicillium setosum, a recently discovered fungus. From this microorganism, six substances belonging to pyranones, diketopiperazines and chlorinated anthraquinones classes were isolated, identified as: 4- methyl-5,6-diihydro-2H-pyran-2-one, fellutanine C, bisdethiobis(methylthio)acetylaranotine, 7-chloroemodin, 2-chloro-1,3,8-trihydroxy-6- (hydroxymethyl)-anthraquinone and 2-chloroemodic acid. Fellutanin C showed promising activity against human leukemia cell line Kasumi-1 as well as selectivity for it. The experiments carried out so far indicate a mechanism of death by apoptosis. Therefore, this work, besides enriching the knowledge of the chemical profile of the genus Swinglea and the fungus P. setosum, offers great prospects for obtaining substances with anti-leukemia action.