RESUMO
Over the years, quinones or its derivatives have been extensively studied due to their broad therapeutic spectrum. However, due to the significant structural differences between the individual naturally occurring quinones, investigation of the precise mechanism of their action is essential. In this context, we have analyzed the mechanism of lapachol [4-hydroxy-3-(3-methylbut-2-enyl)naphthalene-1,2-dione] toxicity using Saccharomyces cerevisiae as eukaryotic model organism. Analyzing yeast (wild type, sod1∆, and gsh1∆) cell growth, we observed a strong cytostatic effect caused by lapachol exposure. Moreover, survival of cells was affected by time- and dose-dependent manner. Interestingly, sod1∆ cells were more prone to lapachol toxicity. In this sense, mitochondrial functioning of sod1∆ cells were highly affected by exposure to this quinone. Lapachol also decreased glutathione (GSH) levels in wild type and sod1∆ cells even though glutathione disulfide (GSSG) remained unchanged. We believe that reduction of GSH contents has contributed to the enhancement of lipid peroxidation and intracellular oxidation, effect much more pronounced in sod1∆ cells. Overall, the collected data suggest that although lapachol can act as an oxidant, it seems that the main mechanism of its action initially consists in alkylation of intracellular targets such as GSH and then generating oxidative stress.
Assuntos
Glutationa/antagonistas & inibidores , Naftoquinonas/farmacologia , Estresse Oxidativo , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/metabolismo , Alquilação , Glutamato-Cisteína Ligase/genética , Glutationa/análise , Peroxidação de Lipídeos , Mitocôndrias/metabolismo , Mutação , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/genética , Superóxido Dismutase-1/genéticaRESUMO
Cobalt(III) complexes are well-suited systems for cytotoxic drug release under hypoxic conditions. Here, we investigate the effect of cytotoxic azide release by cobalt-containing carrier-prototypes for antitumoral prodrugs. In addition, we study the species formed after reduction of Co(3+) â Co(2+) in the proposed models for these prodrugs. Three new complexes, [Co(III)(L)(N3)2]BF4(1), [{Co(II)(L)(N3)}2](ClO4)2(2), and [Co(II)(L)Cl]PF6(3), L=[(bis(1-methylimidazol-2-yl)methyl)(2-(pyridyl-2-yl)ethyl)amine], were synthesized and studied by several spectroscopic, spectrometric, electrochemical, and crystallographic methods. Reactivity and spectroscopic data reveal that complex 1 is able to release N3(-) either after reduction with ascorbic acid, or by ambient light irradiation, in aqueous phosphate buffer (pH6.2, 7.0 and 7.4) and acetonitrile solutions. The antitumoral activities of compounds 1-3 were tested in normoxia on MCF-7 (human breast adenocarcinoma), PC-3 (human prostate) and A-549 (human lung adenocarcinoma epithelial) cell lines, after 24h of exposure. Either complexes or NaN3 presented IC50 values higher than 200 µM, showing lower cytotoxicity than the clinical standard antitumoral complex cisplatin, under the same conditions. Complexes 1-3 were also evaluated in hypoxia on A-549 and results indicate high IC50 data (>200 µM) after 24h of exposure. However, an increase of cancer cell susceptibility to 1 and 2 was observed at 300 µM. Regarding complex 3, no cytotoxic activity was observed in the same conditions. The data presented here indicate that the tridentate ligand L is able to stabilize both oxidation states of cobalt (+3 and +2). In addition, the cobalt(III) complex generates the low cytotoxic cobalt(II) species after reduction, which supports their use as as carrier prototypes for antitumoral prodrugs.
Assuntos
Antineoplásicos/administração & dosagem , Azidas/química , Cloretos/química , Cobalto/química , Pró-Fármacos/administração & dosagem , Antineoplásicos/química , Cristalografia por Raios X , Pró-Fármacos/química , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
There is substantial interest in developing alternative strategies for cancer chemotherapy aiming to increase drug specificity and prevent tumor resistance. Calorie restriction (CR) has been shown to render human cancer cells more susceptible to drugs than normal cells. Indeed, deficiency of nutrient signaling proteins mimics CR, which is sufficient to improve oxidative stress response and life expectancy only in healthy cells. Thus, although CR and reduction of nutrient signaling may play an important role in cellular response to chemotherapy, the full underlying mechanisms are still not completely understood. Here, we investigate the relationship between the nutrient sensor proteins Ras2, Sch9, or Tor1 and the response of calorie-restricted Saccharomyces cerevisiae cells to cisplatin. Using wild-type and nutrient-sensing mutant strains, we show that deletion of any of these proteins mimics CR and is sufficient to increase cell protection. Moreover, we show that glutathione (GSH) is essential for proper CR protection of yeast cells under cisplatin chemotherapy. By measuring the survival rates and GSH levels, we found that cisplatin cytotoxicity leads to a decrease in GSH content reflecting in an increase of oxidative damage. Finally, investigating DNA fragmentation and apoptosis, we conclude that GSH contributes to CR-mediated cell survival.
Assuntos
Cisplatino/toxicidade , Glutationa/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/efeitos dos fármacos , Estresse Fisiológico , Proteínas ras/metabolismo , Apoptose , Fragmentação do DNA , Deleção de Genes , Viabilidade Microbiana/efeitos dos fármacos , Estresse Oxidativo , Fosfatidilinositol 3-Quinases/genética , Proteínas Serina-Treonina Quinases/genética , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas ras/genéticaRESUMO
Our group has recently initiated a study on the development of new prototypes for bioreductive prodrugs, based on Co(III) complexes with the ligand 2,2'-bis(3-hydroxy-1,4-naphthoquinone), H2bhnq. The focus of this work is to investigate the dissociation of bhnq(-2) from the complex upon reduction, and the effects of pH, redox potential, oxygen concentration and nature of the auxiliary ligands on this reaction. The bhnq(2-) ligand is a "non-cytotoxic" agent that was chosen as a probe for the reactivity studies due to its suitable chromophoric properties, at the same time that it resembles more cytotoxic naphthoquinones relevant for cancer therapy. In this way, two Co(III) complexes [Co(bhnq)(L1)]BF4·H2O (1) and [Co(bhnq)(L2)]BF4·H2O (2) (L1=N,N'-bis(pyridin-2-ylmethyl)ethylenediamine and L2=N,N'-dimethyl-N,N'-bis(pyridin-2-ylmethyl)ethylenediamine) were synthesized and fully characterized. The gallium analogs [Ga(bhnq)(L1)]NO3·3H2O (3) and [Ga(bhnq)(L2)]NO3·3H2O (4) were also prepared for helping with the assignments of the redox properties of the cobalt complexes and the structure of 2. Cyclic voltammetry analysis revealed a pH-independent quasi-reversible Co(III)/Co(II) process at -0.22 and -0.08V vs NHE for 1 and 2, respectively. An O2-dependent dissociation of bhnq(2-) was observed for the reaction of 1 with ascorbic acid. For 2, the dissociation of bhnq(2-) was found to be independent on the concentration of O2 and faster than in 1, with little influence of the pH on both complexes. The difference in reactivity between 1 and 2 and their redox properties, among other factors, suggests that 1 undergoes redox cycling, pointed out as a key feature for a prodrug to achieve hypoxic selectivity.
Assuntos
Cobalto/química , Naftoquinonas/química , Pró-Fármacos/química , Simulação por Computador , Cristalografia por Raios X , Gálio/química , Humanos , Concentração de Íons de Hidrogênio , Ligantes , Estrutura Molecular , OxirreduçãoRESUMO
Propolis is a natural product widely used for humans. Due to its complex composition, a number of applications (antimicrobial, antiinflammatory, anesthetic, cytostatic and antioxidant) have been attributed to this substance. Using Saccharomyces cerevisiae as a eukaryotic model we investigated the mechanisms underlying the antioxidant effect of propolis from Guarapari against oxidative stress. Submitting a wild type (BY4741) and antioxidant deficient strains (ctt1∆, sod1∆, gsh1∆, gtt1∆ and gtt2∆) either to 15 mM menadione or to 2 mM hydrogen peroxide during 60 min, we observed that all strains, except the mutant sod1∆, acquired tolerance when previously treated with 25 µg/mL of alcoholic propolis extract. Such a treatment reduced the levels of ROS generation and of lipid peroxidation, after oxidative stress. The increase in Cu/Zn-Sod activity by propolis suggests that the protection might be acting synergistically with Cu/Zn-Sod.
Assuntos
Antioxidantes/farmacologia , Estresse Oxidativo , Própole/farmacologia , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/fisiologia , Brasil , Tolerância a Medicamentos , Peróxido de Hidrogênio/toxicidade , Peroxidação de Lipídeos , Espécies Reativas de Oxigênio/análise , Superóxido Dismutase/análise , /toxicidadeRESUMO
Propolis is a natural product widely used for humans. Due to its complex composition, a number of applications (antimicrobial, antiinflammatory, anesthetic, cytostatic and antioxidant) have been attributed to this substance. Using Saccharomyces cerevisiae as a eukaryotic model we investigated the mechanisms underlying the antioxidant effect of propolis from Guarapari against oxidative stress. Submitting a wild type (BY4741) and antioxidant deficient strains (ctt1, sod1, gsh1, gtt1 and gtt2) either to 15 mM menadione or to 2 mM hydrogen peroxide during 60 min, we observed that all strains, except the mutant sod1, acquired tolerance when previously treated with 25 µg/mL of alcoholic propolis extract. Such a treatment reduced the levels of ROS generation and of lipid peroxidation, after oxidative stress. The increase in Cu/Zn-Sod activity by propolis suggests that the protection might be acting synergistically with Cu/Zn-Sod.(AU)
Assuntos
Própole , Saccharomyces cerevisiae , Estresse Oxidativo , Superóxido Dismutase , AntioxidantesRESUMO
Here we present the synthesis of the dinuclear complex [Cu(II)2(L)Cl3] (1), where L is the deprotonated form of the 3-[(4,7-diisopropyl-1,4,7-triazacyclononan-1-yl)methyl]-2-hydroxy-5-methylbenzaldehyde ligand. The complex was characterized by single crystal X-ray diffraction, potentiometric titration, mass spectrometry, electrochemical and magnetic measurements, EPR, UV-Vis and IR. Complex 1 is able to increase the hydrolysis rate of the diester bis-(2,4-dinitrophenyl)phosphate (2,4-BDNPP) by a factor of 2700, and also to promote the plasmidial DNA cleavage at pH 6 and to inhibit the formazan chromophore formation in redox processes at pH 7. Using Saccharomyces cerevisiae (BY4741) as a eukaryotic cellular model, we observed that 1 presents reduced cytotoxicity. In addition, treatment of wild-type and mutant cells lacking Cu/Zn-superoxide dismutase (Sod1) and cytoplasmic catalase (Ctt1) with 1 promotes increased survival after H2O2 or menadione (O2Ë(-) generator) stress, indicating that 1 might act as a Sod1 and Ctt1 mimetic. Considered together, these results support considerations regarding the dynamic behaviour of an unsymmetrical dinuclear copper(II) complex in solid state and in aqueous pH-dependent solution.
Assuntos
Complexos de Coordenação/química , Cobre/química , Compostos Heterocíclicos/química , Antioxidantes/química , Antioxidantes/metabolismo , Materiais Biomiméticos/síntese química , Materiais Biomiméticos/metabolismo , Materiais Biomiméticos/farmacologia , Catálise , Complexos de Coordenação/metabolismo , Complexos de Coordenação/farmacologia , Cristalografia por Raios X , DNA/química , DNA/metabolismo , Clivagem do DNA , Concentração de Íons de Hidrogênio , Cinética , Ligantes , Magnetismo , Conformação Molecular , Saccharomyces cerevisiae/efeitos dos fármacos , Superóxido Dismutase/metabolismo , TemperaturaRESUMO
Dimerization of lawsone occurs upon reaction with Co(BF(4))(2)·6H(2)O and N,N'-bis(pyridin-2-ylmethyl)ethylenediamine (py(2)en) to produce the mononuclear complex [Co(III)(bhnq)(py(2)en)]BF(4)·H(2)O (1). This complex has been investigated as a prototype of a bioreductive prodrug, where the bhnq(2-) ligand acts as a model for cytotoxic naphthoquinones. Cyclic voltammetry data in aqueous solution have shown a quasi-reversible Co(III)/Co(II) process at E(1/2) = -0.26 V vs Fc/Fc(+). Reactivity studies revealed the dissociation of bhnq(2-) from the complex upon reduction of 1 with ascorbic acid, and a dependence of the reaction rate on the oxygen concentration suggests the occurrence of redox cycling.
Assuntos
Cobalto/química , Naftoquinonas/química , Compostos Organometálicos/farmacologia , Pró-Fármacos/síntese química , Pró-Fármacos/farmacologia , Saccharomyces cerevisiae/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Cristalografia por Raios X , Dimerização , Modelos Moleculares , Estrutura Molecular , Compostos Organometálicos/síntese química , Compostos Organometálicos/química , Oxirredução , Pró-Fármacos/química , Saccharomyces cerevisiae/citologia , Relação Estrutura-AtividadeRESUMO
Propolis is a natural product widely used for humans. Due to its complex composition, a number of applications (antimicrobial, antiinflammatory, anesthetic, cytostatic and antioxidant) have been attributed to this substance. Using Saccharomyces cerevisiae as a eukaryotic model we investigated the mechanisms underlying the antioxidant effect of propolis from Guarapari against oxidative stress. Submitting a wild type (BY4741) and antioxidant deficient strains (ctt1Δ, sod1Δ, gsh1Δ, gtt1Δ and gtt2Δ) either to 15 mM menadione or to 2 mM hydrogen peroxide during 60 min, we observed that all strains, except the mutant sod1Δ, acquired tolerance when previously treated with 25 µg/mL of alcoholic propolis extract. Such a treatment reduced the levels of ROS generation and of lipid peroxidation, after oxidative stress. The increase in Cu/Zn-Sod activity by propolis suggests that the protection might be acting synergistically with Cu/Zn-Sod.
Assuntos
Antioxidantes/farmacologia , Estresse Oxidativo , Própole/farmacologia , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/fisiologia , Brasil , Tolerância a Medicamentos , Peróxido de Hidrogênio/toxicidade , Peroxidação de Lipídeos , Espécies Reativas de Oxigênio/análise , Superóxido Dismutase/análise , Vitamina K 3/toxicidadeRESUMO
Two flavonoids 3,5,7,3',4'-pentahydroxy-6-prenylflavonol (1) and 3,5,7,3',4'-pentahydroxy-8-methyl-6-prenylflavonol (2) were isolated from the ethyl acetate extract of sheaths of Vellozia kolbekii Alves (Velloziaceae). This is the first time that compound 2 has been described. The crude extract and flavonoids were found to be active as 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavengers and were able to the increase tolerance of the eukaryotic microorganism Saccharomyces cerevisiae to oxidative stress generated by H(2)O(2). The protective effect was correlated with a reduction in the oxidation of proteins and lipids. In addition, flavonoids isolated from Velloziaceae showed an inhibitory effect on mutations in p53, which is mutated and nonfunctional in more than 50% of cases of human cancer.