RESUMO
A series of 1,2,4-triazole derivatives were synthesized and assigned as potential anti-tuberculosis substances. The molecular and crystal structures for the model compounds C1, C12, and C13 were determined using X-ray analysis. The X-ray investigation confirmed the synthesis pathway and the assumed molecular structures for analyzed 1,2,4-triazol-5-thione derivatives. The conformational preferences resulting from rotational degrees of freedom of the 1,2,4-triazole ring substituents were characterized. The lipophilicity (logP) and electronic parameters as the energy of frontier orbitals, dipole moments, NBO net charge distribution on the atoms, and electrostatic potential distribution for all structures were calculated at AM1 and DFT/B3LYP/6-311++G(d,p) level. The in vitro test was done against M. tuberculosis H37Ra, M. phlei, M. smegmatis, and M. timereck. The obtained results clearly confirmed the antituberculosis potential of compound C4, which turned out to be the most active against Mycobacterium H37Ra (MIC = 0.976 µg/mL), Mycobaterium pheli (MIC = 7.81 µg/mL) and Mycobacerium timereck (62.6 µg/mL). Satisfactory results were obtained with compounds C8, C11, C14 versus Myc. H37Ra, Myc. pheli, Myc. timereck (MIC = 31.25-62.5 µg/mL). The molecular docking studies were carried out for all investigated compounds using the Mycobacterium tuberculosis cytochrome P450 CYP121 enzyme as molecular a target connected with antimycobacterial activity.
Assuntos
Antituberculosos/farmacologia , Triazóis/farmacologia , Testes de Sensibilidade Microbiana/métodos , Simulação de Acoplamento Molecular/métodos , Mycobacterium tuberculosis/efeitos dos fármacos , Relação Estrutura-AtividadeRESUMO
A series of thiosemicarbazide derivatives was designed and synthesized by reaction of carboxylic acid hydrazide with isothiocyanates. The molecular structures of the investigated thiosemicarbazides were confirmed and characterized by spectroscopic analysis. The conformational preference of carbonylthiosemicarbazide chain and intra- and intermolecular interactions in the crystalline state were characterized using X-ray analysis. The antituberculosis activity of the target compounds were tested in vitro against four Mycobacterium strains: M. H37Ra, M. phlei, M. smegmatis, M. timereck. The most active compounds were those with 2-pyridine ring. They exhibited lower minimal inhibitory concentration (MIC) values in the range 7.81â»31.25 µg/mL in comparison to the other isomers. Compound 5 had activity against M. smegmatis at a concentration of 7.81 µg/mL whereas compound 2 had activity against all tested strains at a concentration of 15.625 µg/mL. The molecular docking studies were performed for investigated compounds using the Mycobacterium tuberculosis glutamine synthetase MtGS as their molecular target.
Assuntos
Antituberculosos/química , Antituberculosos/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Simulação de Acoplamento Molecular , Relação Quantitativa Estrutura-Atividade , Semicarbazidas/química , Semicarbazidas/farmacologia , Antituberculosos/síntese química , Sítios de Ligação , Ligação de Hidrogênio , Modelos Moleculares , Simulação de Dinâmica Molecular , Estrutura Molecular , Ligação Proteica , Semicarbazidas/síntese químicaRESUMO
The most commonly used plant source of ß-elemene is Curcuma wenyujin Y. H. Chen & C. Ling (syn. of Curcuma aromatic Salisb.) with its content in supercritical CO2 extract up to 27.83%. However, the other rich source of this compound is Nigella damascena L. essential oil, in which ß-elemene accounts for 47%. In this work, the effective protocol for preparative isolation of ß-elemene from a new source-N. damascena essential oil-using high performance counter-current chromatography HPCCC was elaborated. Furthermore, since sesquiterpens are known as potent antimicrobials, the need for finding new agents designed to combat multi-drug resistant strains was addressed and the purified target compound and the essential oil were tested for its activity against a panel of Gram-positive and Gram-negative bacteria, fungi, and mycobacterial strains. The application of the mixture of petroleum ether, acetonitrile, and acetone in the ratio 2:1.5:0.5 (v/v) in the reversed phase mode yielded ß-elemene with high purity in 70 min. The results obtained for antimicrobial assay clearly indicated that N. damascena essential oil and isolated ß-elemene exert action against Mycobacterium tuberculosis strain H37Ra.
Assuntos
Anti-Infecciosos , Mycobacterium tuberculosis/crescimento & desenvolvimento , Nigella damascena/química , Óleos Voláteis/química , Sesquiterpenos , Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Testes de Sensibilidade Microbiana , Sesquiterpenos/química , Sesquiterpenos/farmacologiaRESUMO
Background: On account of emergence of multi- and extensively drug-resistant Mycobacterium tuberculosis (Mtb) strains, combinations of drugs with natural compounds were tested to search for antibiotic activity enhancers. In this work we studied terpenes (α-pinene, bisabolol, ß-elemene, (R)-limonene, (S)-limonene, myrcene, sabinene), which are the main constituents of essential oil obtained from Mutellina purpurea L., a plant with described antitubercular activity, to investigate their interactions with antibiotics against reference Mtb strains and multidrug-resistant clinical isolates. Methods: The serial dilution method was used to evaluate the minimal inhibitory concentration (MIC) of tested compounds, while the fractional inhibitory concentration index (FICI) was calculated for characterization of interactions. Moreover, IC50 values of tested compounds were determined using monkey kidney epithelial cell line (GMK). Results: The combinations of all studied terpenes with ethambutol or rifampicin resulted in a synergistic interaction. Bisabolol and (R)-limonene decreased the MIC for rifampicin at least two-fold for all tested strains, however no synergistic action was observed against virulent strains. The tested terpenes showed slight (bisabolol) or no cytotoxic effect against normal eukaryotic cells in vitro. Conclusions: The obtained enhanced activity (FICI < 0.5) of ethambutol and rifampicin against H37Ra strain under the influence of the studied terpenes may be correlated to the capability of essential oil constituents to modify bacterial resistance mechanisms in general. The observed differences in avirulent and virulent bacteria susceptibility to terpenes tested separately and in combinations with antibiotics can be correlated with the differences in the cell wall structure between H37Ra mutant and all virulent strains.
Assuntos
Antituberculosos/farmacologia , Produtos Biológicos/farmacologia , Mycobacterium tuberculosis/efeitos dos fármacos , Terpenos/farmacologia , Antituberculosos/química , Produtos Biológicos/química , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Sinergismo Farmacológico , Humanos , Concentração Inibidora 50 , Testes de Sensibilidade Microbiana , Mycobacterium tuberculosis/genética , Terpenos/químicaRESUMO
We present the specific cooperative effect of a semisynthetic glycopeptide antibiotic teicoplanin and chiral ionic liquids containing the (1R,2S,5R)-(-)-menthol moiety on the chiral recognition of enantiomers of mandelic acid, vanilmandelic acid, and phenyllactic acid. Experiments were performed chromatographically on an Astec Chirobiotic T chiral stationary phase applying the mobile phase with the addition of the chiral ionic liquids. The stereoselective binding of enantiomers to teicoplanin in presence of new chiral ionic liquids were evaluated applying thermodynamic measurements and the docking simulations. Both the experimental and theoretical methods revealed that the chiral recognition of enantiomers in the presence of new chiral ionic liquids was enthalpy driven. The changes of the teicoplanin conformation occurring upon binding of the chiral ionic liquids are responsible for the differences in the standard changes in Gibbs energy (ΔG0 ) values obtained for complexes formed by the R and S enantiomers and teicoplanin. Docking simulations revealed the steric adjustment between the chiral ionic liquids cyclohexane ring (chair conformation) and the ß-d-glucosamine ring of teicoplanin and additionally hydrophobic interactions between the decanoic aliphatic chain of teicoplanin and the alkyl group of the tested salts. The obtained terpene derivatives can be considered as "structural task-specific ionic liquids" responsible for enhancing the chiral resolution in synergistic systems with two chiral selectors.
Assuntos
Líquidos Iônicos/química , Teicoplanina/química , Terpenos/química , Cromatografia Líquida de Alta Pressão , EstereoisomerismoRESUMO
OBJECTIVE: In this study, we aimed to describe the influence of natural terpenes on the antimycobacterial activity of first-line tuberculostatic drugs against isolated Mycobacterium tuberculosis. MATERIALS AND METHODS: The natural terpenes used in this study were R-limonene, S-limonene, myrcene, sabinene, α-pinene, and ß-elemene. The values of the minimum inhibitory concentration (MIC) for these terpenes, as well as for combinations of terpenes with tuberculostatic antibiotics (ethambutol, isoniazid, and rifampicin), were determined using a tube log2 dilution method in the range of 125-0.059 µg/mL. RESULTS: S-limonene had a strong synergistic effect with all tested antibiotics (MIC decreased from 16 to 0.475 µg/mL for ethambutol, from 16 to 0.237 µg/mL for rifampicin, and from 32 to 0.475 µg/mL for isoniazid). Combinations of myrcene, R-limonene, ß-elemene, and sabinene with tuberculostatic antibiotics resulted in a decreased MIC of the antibiotics (from 3.9 to 0.475 µg/mL for ethambutol, from 15 to 0.475 µg/mL for isoniazid, and from 0.475 to 0.237 µg/mL for rifampicin) while combinations of α-pinene with ethambutol and isoniazid resulted in increased MIC values (from 16 to 125 µg/mL for ethambutol, and from 32 to 125 µg/mL for isoniazid). Rifampicin had a synergistic increase in activity with all the tested compounds. CONCLUSIONS: Our study showed that terpenes enhance the activity of tuberculostatic antibiotics.
Assuntos
Antituberculosos/farmacologia , Mycobacterium tuberculosis/efeitos dos fármacos , Terpenos/farmacologia , Monoterpenos Acíclicos , Alcenos/farmacologia , Monoterpenos Bicíclicos , Cicloexenos/farmacologia , Relação Dose-Resposta a Droga , Quimioterapia Combinada , Etambutol/farmacologia , Humanos , Isoniazida/farmacologia , Limoneno , Testes de Sensibilidade Microbiana , Monoterpenos/farmacologia , Rifampina/farmacologia , Sesquiterpenos/farmacologiaRESUMO
OBJECTIVE: The aim of this study was to evaluate the antimycobacterial activity of the essential oil (EO) of Mutellina purpurea L. and its main constituents against the M. tuberculosis H37Ra strain. MATERIALS AND METHODS: The M. purpurea EO was obtained by hydrodistillation, while its main constituents were purchased. The minimal inhibitory concentration values were determined by the log2 dilution method. Visualization of the effects of the tested substances on M. tuberculosis was performed using a transmission electron microscope (TEM). Mathematical shape descriptors such as area, circularity, aspect ratio and roundness were calculated to describe morphological changes in bacterial cell shape. RESULTS: The EO of M.purpurea and all substances tested in this experiment showed a significant antimycobacterial activity. The most active was α-pinene followed by bisabolol and myrcene (8, 16 and 32 µg/ml, respectively). The EO and limonene exhibited the same antimicrobial activity (64 µg/ml). The TEM images and shape descriptors showed significant changes in the overall tuberculosis cell shape and cytoplasm homogeneity (uniformity and consistency) CONCLUSIONS: In this study, the low molecular weight compounds of mono- and sesquiterpenes penetrated/destabilized the complex mycobacterial cell wall and decreased its viability. There is a need for further experiments to explain the mechanism of action of these small particles.
Assuntos
Antibacterianos/farmacologia , Apiaceae , Forma Celular/efeitos dos fármacos , Citoplasma/efeitos dos fármacos , Mycobacterium tuberculosis/efeitos dos fármacos , Óleos Voláteis/farmacologia , Antibacterianos/química , Monoterpenos Bicíclicos , Cicloexenos/farmacologia , Relação Dose-Resposta a Droga , Humanos , Limoneno , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Transmissão , Sesquiterpenos Monocíclicos , Monoterpenos/farmacologia , Óleos Voláteis/química , Sesquiterpenos/farmacologia , Terpenos/farmacologiaRESUMO
The global threat of tuberculosis demands the search for alternative antimycobacterial drugs. The present study examined roots and petioles from Rheum rhaponticum for antimycobacterial activity. Crude methanol extracts and eight phenolic compounds isolated by preparative column chromatography were tested against Mycobacterium tuberculosis H37Ra and M. bovis using the broth dilution method. The extract from roots and its components, such as rhaponticin, deoxyrhaponticin, resveratrol, barbaloin, aloe-emodin, and chrysophanol were found to have antimycobacterial activity against both microorganisms. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration of all the investigated samples ranged from 32 to 512 microg/mL. The anthracene derivatives were the most active; their MICs were 32, 64, and 64 microg/mL (M. tuberculosis H37Ra) and 128, 64, and 64 microg/mL (M. bovis), respectively. The microorganisms were resistant to stimulation with extract from petioles, as were quercetin and rutin. The results showed that anthracene and stilbene derivatives play a prominent role in the antimycobacterial properties of R. rhaponticum.
Assuntos
Antibacterianos/farmacologia , Mycobacterium tuberculosis/efeitos dos fármacos , Extratos Vegetais/farmacologia , Rheum/química , Antibacterianos/análise , Testes de Sensibilidade Microbiana , Mycobacterium bovis/efeitos dos fármacos , Extratos Vegetais/análise , Raízes de Plantas/químicaRESUMO
In the past few years, there has been a significant increase in detection of drug resistant strains of Mycobacterium tuberculosis. Search for new antimycobacterial drugs brought natural sources with their chemical diversity in focus. Especially essential oils, produced by plants also for toxic effect, are reservoir of potentially antitubercular compounds. In the present work, we exposed M. tuberculosis H37Ra ATCC 25177 strain to some terpenes commonly occurring in essential oils. Gene expression profiling was used to explore possible influence of these compounds on stress sensing and envelope preserving function. Expression of two genes dprE1 involved in cell wall synthesis and clgR responsible for regulation of cell membrane preservation was investigated. We report that two out of five tested compounds: ß-elemene and R-limonene alter expression of dprE1 and clgR genes. These findings indicate various mechanisms of action of essential oils compounds on M. tuberculosis. Especially the clgR expression seemed to be the perfect marker of stress sensing and envelope preserving systems status.
Assuntos
Oxirredutases do Álcool/genética , Antituberculosos/farmacologia , Proteínas de Bactérias/genética , Cicloexenos/farmacologia , Mycobacterium tuberculosis/genética , Óleos Voláteis/farmacologia , Sesquiterpenos/farmacologia , Terpenos/farmacologia , Biomarcadores/metabolismo , Perfilação da Expressão Gênica , Genes Bacterianos , Limoneno , Testes de Sensibilidade Microbiana , Mycobacterium tuberculosis/efeitos dos fármacos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Transcrição Gênica/efeitos dos fármacosRESUMO
As a continuation of previous tests concerning new N-substituted 3-amino-4-phenyl-5-oxo-pyrazolinecarboxamide derivatives (R3, R4 and R8) of notable antibacterial activity, their antitubercular potential against different mycobacterial strains was estimated. Tests performed on virulent (reference and clinical) strains of Mycobacterium bovis and Mycobacterium tuberculosis revealed the highest therapeutic potential of R8 derivative: MIC within the range 7.8-15.6⯵g/ml and TI (therapeutic index) within the range 46.5-93. Moreover, the synergistic interaction was found between R3, R4 and R8 derivatives and rifampicin, one of the front-line antitubercular drugs. R8/rifampicin mixture in concentrations effective in inhibition of Mycobacterium tuberculosis strain was non-cytotoxic against GMK cells, displaying cell viability approximately 88-97% when compared to control. Molecular docking study enabled to conclude that enoyl acyl carrier protein reductase (InhA) can be considered as a potential molecular target of tested pyrazole derivatives. Although further modifications of chemical structure of the investigated pyrazole derivatives is required, in order to increase their antitubercular efficacy and therapeutic safety, these compounds, in particular R8 compound, can be promising for the treatment of human and bovine tuberculosis.