Atividade antibacteriana de híbridos curcumina-cinamaldeído contra células planctônicas e biofilmes de MSSA e MRSA
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2019-06-06
Autores
Marques, Beatriz de Carvalho
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Universidade Estadual Paulista (Unesp)
Resumo
Staphylococcus aureus é uma bactéria Gram-positiva, comensal e patogênica, que coloniza aproximadamente 30% da população humana, apresentando resistência a vários antibióticos. Estudos indicam a bioatividade da curcumina e do cinamaldeído contra S. aureus, no entanto, ambos apresentam limitações que impedem seu uso como agente terapêutico. Assim, a fim de otimizar a atividade anti-S. aureus da curcumina e do cinamaldeído, bem como suas propriedades químico-biológicas, no presente estudo, duas séries de híbridos curcumina-cinamaldeído foram planejadas e sintetizadas, sendo posteriormente avaliadas contra S. aureus sensível e resistente à meticilina (MSSA e MRSA). O híbrido mais ativo foi selecionado para avaliação da sua combinação com antibióticos, e quanto a sua atividade anti-adesão, antibiofilme e no tempo de morte de MSSA e MRSA. A toxicidade in vivo deste hídrido também foi avaliada utilizando o invertebrado Galleria mellonella. Os híbridos ainda foram avaliados quanto à sua atividade antibacteriana contra outras espécies bacterianas e Mycobacterium tuberculosis, e investigados in silico quanto às suas propriedades drug-likeness. Trinta híbridos foram obtidos com pureza entre 86 e 100%, sendo doze inéditos. Em geral, a hibridação molecular entre a curcumina e o cinamaldeído manteve ou aumentou a atividade antibacteriana dos híbridos quando comparados com a curcumina e com o cinamaldeído, sendo que a lipofilicidade foi um parâmetro central na bioatividade contra MSSA e MRSA. O híbrido 20 apresentou atividade anti-S. aureus mais pronunciada, com valores de Concentração Inibitória Mínima (CIM) de 7,8 μg/mL e Concentração Bactericida Mínima (CBM) de 15,6 μg/mL para MSSA, e CIM de 3,9 μg/mL e CBM de 7,8 μg/mL para MRSA, sendo selecionado para os ensaios subsequentes. As associações entre 20 e vancomicina ou meticilina aumentaram a atividade antibacteriana contra MSSA e MRSA. O tratamento com 20 diminuiu a aderência das cepas de MSSA e MRSA aos queratinócitos humanos, quando comparado ao grupo não tratado. Para os biofilmes, após o tratamento com 20, verificou-se uma diminuição na sua sobrevivência quando comparado ao controle negativo, apresentando atividade antibiofilme similar ou superior à vancomicina e à meticilina. No ensaio de tempo de morte, o tratamento com 20 diminuiu a população bacteriana, indicando ação bactericida. Além disso, 20 apresentou toxicidade a 90 mg/kg, após 72 horas de tratamento, sendo responsável pela morte de 50% da larvas de Galleria mellonella. Os híbridos mais lipofílicos demonstraram maior bioatividade contra Enterococcus faecalis, Staphylococcus epidermidis e Pseudomonas aeruginosa. Os híbridos não apresentaram atividade antibacteriana contra Escherichia coli. Acinetobacter baumannii foi a espécie bacteriana mais suscetível aos híbridos. Contra Mycobacterium tuberculosis, verificou-se que a inserção de grupos retiradores de elétrons e lipofílicos aumentou a atividade antimicobacteriana. Com relação aos estudos in silico, os híbridos não violaram as regras estabelecidas por Lipinski e Veber, corroborando boas propriedades drug-likeness. Neste estudo, pode-se comprovar a atividade antibacteriana de híbridos de curcumina-cinamaldeído, estimulando o estudo dessas substâncias, a fim de buscar agentes capazes de atuar contra cepas resistentes.
Staphylococcus aureus is a gram-positive, commensal and pathogenic bacteria that colonizes approximately 30% of the human population, presenting resistance to several antibiotics. Studies indicate the bioactivity of curcumin and cinnamaldehyde against S. aureus, however, both have limitations that prevent its use as a therapeutic agent. Thus, in order to optimize anti-S. aureus activity of curcumin and cinnamaldehyde, as well as its chemical-biological properties, in the present study, two series of curcumincinnamaldehyde hybrids were planned and synthesized and subsequently evaluated against methicillin-susceptible and methicillin-resistant S. aureus (MSSA and MRSA). The most active hybrid was selected for evaluation of its combination with antibiotics, and for its anti-adhesion activity, antibiofilm and the time kill of MSSA and MRSA. In vivo toxicity of this hybrid was also evaluated using the Galleria mellonella invertebrate. Hybrids were further evaluated for their antibacterial activity against other bacterial species and Mycobacterium tuberculosis, and investigated in silico for their drug-likeness properties. Thirty hybrids were obtained with purity between 86 and 100%, of which twelve were unpublished. In general, the molecular hybridization between curcumin and cinnamaldehyde maintained or increased the antibacterial activity of hybrids when compared to curcumin and cinnamaldehyde, and lipophilicity was a central parameter in bioactivity against MSSA and MRSA. Hybrid 20 showed more pronounced activity, presented Minimal Inhibitory Concentration (MIC) of 7.8 μg/mL and Minimum Bactericidal Concentration (MBC) of 15.6 μg/mL for MSSA, and MIC of 3.9 μg/mL and MBC of 7,8 μg/mL for MRSA, being selected for subsequent assays. Associations between 20 and vancomycin or methicillin increased the antibacterial activity against MSSA and MRSA. Treatment with 20 decreased the adherence of MSSA and MRSA strains onto human keratinocytes when compared to the untreated group. For biofilms, after treatment with 20, there was a decrease in their survival when compared to the negative control, presenting antibiofilm activity similar or superior to vancomycin and methicillin. In time kill assay, treatment with 20 decreased the bacterial population, indicating bactericidal action. In addition, 20 presented toxicity at 90 mg/kg, after 72 hours of treatment, being responsible for the death of 50% of G. mellonella. Against Enterococcus faecalis, Staphylococcus epidermidis and Pseudomonas aeruginosa, the more lipophilic hybrids demonstrated greater bioactivity. Hybrids showed no antibacterial activity against Escherichia coli. Acinetobacter baumannii was the bacterial species most susceptible to hybrids. Against Mycobacterium tuberculosis, it was found that the insertion of electron withdrawing and lipophilic groups increased the antimycobacterial activity. In relation to in silico studies, hybrids did not violate the rules established by Lipinski and Veber, corroborating good drug-likeness properties. In this study, we can prove the antibacterial activity of curcumin-cinnamaldehyde hybrids, stimulating the study of these substances, in order to find agents capable of acting against resistant strains.
Staphylococcus aureus is a gram-positive, commensal and pathogenic bacteria that colonizes approximately 30% of the human population, presenting resistance to several antibiotics. Studies indicate the bioactivity of curcumin and cinnamaldehyde against S. aureus, however, both have limitations that prevent its use as a therapeutic agent. Thus, in order to optimize anti-S. aureus activity of curcumin and cinnamaldehyde, as well as its chemical-biological properties, in the present study, two series of curcumincinnamaldehyde hybrids were planned and synthesized and subsequently evaluated against methicillin-susceptible and methicillin-resistant S. aureus (MSSA and MRSA). The most active hybrid was selected for evaluation of its combination with antibiotics, and for its anti-adhesion activity, antibiofilm and the time kill of MSSA and MRSA. In vivo toxicity of this hybrid was also evaluated using the Galleria mellonella invertebrate. Hybrids were further evaluated for their antibacterial activity against other bacterial species and Mycobacterium tuberculosis, and investigated in silico for their drug-likeness properties. Thirty hybrids were obtained with purity between 86 and 100%, of which twelve were unpublished. In general, the molecular hybridization between curcumin and cinnamaldehyde maintained or increased the antibacterial activity of hybrids when compared to curcumin and cinnamaldehyde, and lipophilicity was a central parameter in bioactivity against MSSA and MRSA. Hybrid 20 showed more pronounced activity, presented Minimal Inhibitory Concentration (MIC) of 7.8 μg/mL and Minimum Bactericidal Concentration (MBC) of 15.6 μg/mL for MSSA, and MIC of 3.9 μg/mL and MBC of 7,8 μg/mL for MRSA, being selected for subsequent assays. Associations between 20 and vancomycin or methicillin increased the antibacterial activity against MSSA and MRSA. Treatment with 20 decreased the adherence of MSSA and MRSA strains onto human keratinocytes when compared to the untreated group. For biofilms, after treatment with 20, there was a decrease in their survival when compared to the negative control, presenting antibiofilm activity similar or superior to vancomycin and methicillin. In time kill assay, treatment with 20 decreased the bacterial population, indicating bactericidal action. In addition, 20 presented toxicity at 90 mg/kg, after 72 hours of treatment, being responsible for the death of 50% of G. mellonella. Against Enterococcus faecalis, Staphylococcus epidermidis and Pseudomonas aeruginosa, the more lipophilic hybrids demonstrated greater bioactivity. Hybrids showed no antibacterial activity against Escherichia coli. Acinetobacter baumannii was the bacterial species most susceptible to hybrids. Against Mycobacterium tuberculosis, it was found that the insertion of electron withdrawing and lipophilic groups increased the antimycobacterial activity. In relation to in silico studies, hybrids did not violate the rules established by Lipinski and Veber, corroborating good drug-likeness properties. In this study, we can prove the antibacterial activity of curcumin-cinnamaldehyde hybrids, stimulating the study of these substances, in order to find agents capable of acting against resistant strains.
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Palavras-chave
Staphylococcus aureus, Curcumina, Cinalmaldeído, Hibridação molecular, Curcumin, Cinalmaldehyde, Molecular hybridization