RESUMEN
The aim of this investigation was to measure the activity of four 4-hydroxycoumarin derivatives - three of them were described before and one was newly synthesized. The substances were ethyl 2-[(4-hydroxy-2-oxo-2H-chromen-3-yl)(4-hydroxyphenyl)methyl]-3-oxobutanoate (SS-14), 4-[1-(4-hydroxy-2-oxo-2H-chromen-3-yl)-2-(ethoxycarbonyl)-3-oxobutyl]benzoic acid (SS-17), ethyl 2-[(4-hydroxy-2-oxo-2H-chromen-3-yl)(3-nitrophenyl)methyl]-3-oxobutanoate (SS-21) and ethyl 2-[(3,4,5-trimethoxyphenyl)(4-hydroxy-2-oxo-2H-chromen-3-yl)methyl]-3-oxobutanoate (T-2). The synthesis of T-2 consists of two steps. First step was Knoevenagel reaction between 3,4,5-trimethoxybenzaldehyde and ethylacetoacetate. Ethyl 2-(3,4,5-trimethoxy)-phenylmethyleneacetoacetate was the product. Second step was Michael addition reaction between the latter product and 4-hydroxycoumarin. All the compounds were tested in vitro for antioxidant activity in hypochlorous system. The assay was based on the luminol-dependent chemiluminescence of free radicals, which decreased in the presence of 4-hydroxycoumarin derivative. Compound SS-14 (ethyl 2-[(4-hydroxy-2-oxo-2H-chromen-3-yl)(4-hydroxyphenyl)methyl]-3-oxobutanoate) expresses the best scavenger activity at the highest concentration (10(-4)mol/L).
Asunto(s)
4-Hidroxicumarinas/química , 4-Hidroxicumarinas/farmacología , Diseño de Fármacos , Depuradores de Radicales Libres/química , Depuradores de Radicales Libres/farmacología , 4-Hidroxicumarinas/síntesis química , Depuradores de Radicales Libres/síntesis química , Humanos , Luminiscencia , Relación Estructura-ActividadRESUMEN
The present study describes the application of several chemiluminescent (CL) methods for evaluation of antioxidant and immunomodulation effects of psychotropic drugs upon phagocytes: KO2-induced luminal-dependent CL for detection of superoxide anion radicals in a pure chemical system; PMA- and A23187-induced CL of peritoneal macrophages for detection of free radicals in cell suspension; and CL, produced by the luciferase-catalyzed luciferin + ATP reaction, for evaluation of cell viability before and after drug application. These methods provide also a way to investigate the location of drug action. It was found that the psychotropic drugs in fluence the 'oxidative burst' of macrophages through two mechanisms: by expression of drug antioxidant properties and/or by a direct immunomodulation effect.
Asunto(s)
Antioxidantes/farmacología , Factores Inmunológicos/farmacología , Mediciones Luminiscentes/métodos , Macrófagos Peritoneales/inmunología , Macrófagos Peritoneales/metabolismo , Psicotrópicos/farmacología , Amitriptilina/farmacología , Animales , Antidepresivos Tricíclicos/farmacología , Antipsicóticos/farmacología , Calcimicina/farmacología , Calmodulina/metabolismo , Supervivencia Celular/efectos de los fármacos , Clorpromazina/farmacología , Clorprotixeno/farmacología , Imipramina/farmacología , Técnicas In Vitro , Indicadores y Reactivos , Ionóforos/farmacología , Luminol/química , Activación de Macrófagos/efectos de los fármacos , Macrófagos Peritoneales/efectos de los fármacos , Masculino , Ratas , Acetato de Tetradecanoilforbol/farmacología , Tioxantenos/farmacologíaRESUMEN
The antioxidant properties of galantamine hydrobromide ((4alpha,6beta)-4a,5,9,10,11,12-hexahydro-3-methoxy-11-methyl-6H-benzofuro[3a,3,2-ef][2]benzazepin-6-ol hydrobromide) were studied in vitro, using luminol-dependent chemiluminescence and spectrophotometry. It was found that this compound was a scavenger of reactive oxygen species (ROS). By comparing the antioxidant effects of galantamine ((4alpha,6beta)-4a,5,9,10,11,12-hexahydro-3-methoxy-11-methyl-6H-benzofuro[3a,3,2-ef][2]benzazepin-6-ol), galantamine hydrobromide, narwedine (4a,5,9,10,11,12-hexahydro-3-methoxy-11-methyl-6H-benzofuro[3a,3,2-ef][2]benzazepin-6-one), and narwedine hydrobromide it was found that the antioxidant activity depended on the enolic OH group in the molecule. The presence of a quaternary nitrogen in the compound increased the strength of the scavenging effect. It is proposed that the antioxidant properties observed in vitro may contribute to the therapeutical effect of galantamine hydrobromide on patients with brain degeneration.
Asunto(s)
Antioxidantes/farmacología , Galantamina/química , Galantamina/farmacología , Depuradores de Radicales Libres , Galantamina/síntesis química , Peróxido de Hidrógeno , Radical Hidroxilo/análisis , Luminol , Espectroscopía de Resonancia Magnética , Oxidación-Reducción , SuperóxidosRESUMEN
The effects of some phenothiazines (promethazine, PMZ; chlorpromazine, CPZ; levomepromazine, LVPZ; thioridazine, TRDZ; trifluoperazine, TFPZ) on the activation and viability of rat peritoneal macrophages were investigated. The macrophage activation was estimated by measuring of luminol-dependent chemiluminescence, induced by phorbol-12-myristate-13-acetate (PMA) (a protein kinase C activator) or calcium ionophore A23187. The viability of macrophages was determined using ATP bioluminescence as a criterion of cell viability. It was observed that all drugs, in concentrations higher than 1 micromol/L, markedly decreased the chemiluminescent index of PMA-activated or A23187-activated macrophages. The inhibitory effect was dose-dependent. It was better expressed in the case of CPZ, followed by TFPZ and TRDZ, and less expressed in the case of PMZ and LVPZ. The suppression of chemiluminescence of PMA-/A23187-activated macrophages by phenothiazines was not a result of their cytotoxic effect. Moreover, it was found that all drugs dose-dependently enhanced the viability of macrophages, estimated by ATP production. The inhibitory effects of phenothiazines on the chemiluminescence of PMA-/A23187-activated macrophages were greater than their ability to decrease KO2-induced chemiluminescence as a result of interaction with superoxide radicals. It may be supposed that the inhibitory effect of phenothiazines on PMA-/A23187-induced chemiluminescence of macrophages is a result not only of interaction between drugs and superoxide radicals, generated during the "oxidative burst" of activated cells. Presumably the drugs have an immunomodulating effect on rat peritoneal macrophages.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Calcio/metabolismo , Activación de Macrófagos/efectos de los fármacos , Macrófagos Peritoneales/efectos de los fármacos , Fenotiazinas/farmacología , Proteína Quinasa C/metabolismo , Adyuvantes Inmunológicos/química , Animales , Calcimicina/farmacología , Supervivencia Celular/efectos de los fármacos , Activadores de Enzimas/farmacología , Técnicas In Vitro , Activación de Macrófagos/inmunología , Macrófagos Peritoneales/enzimología , Macrófagos Peritoneales/inmunología , Macrófagos Peritoneales/metabolismo , Masculino , Estructura Molecular , Fenotiazinas/química , Ratas , Ratas WistarRESUMEN
Influenza virus infection is associated with development of oxidative stress in lung and blood plasma, viz. increase of primary and secondary lipid peroxidation products. It was established that rimantadine treatment led to a decrease of the products of lipid peroxidation in tissues of mice experimentally infected with influenza virus A/Aichi/2/68 (H3N2). The effect is strongest in blood plasma (a decrease of about 50%) and weaker in the lung (about 20%). To elucidate the mechanism of this action of rimantadine, experiments were carried out with some model systems. The capability of rimantadine to scavenge superoxide radicals (scavenging properties) was studied in a system of xanthine-xanthine oxidase to generate superoxide. The amount of superoxide was measured spectrophotometrically by the NBT-test and chemiluminesce. Rimantadine does not show scavenging properties and its antioxidant effect observed in vivo, is not a result of its direct action on the processes of lipid peroxidation and/or interaction with antioxidant enzymes. The antioxidant properties of rimantadine were investigated by measurement of induced lipid peroxidation in a Fe2+ and (Fe2+ - EDTA) system with an egg liposomal suspension. Our findings with model systems do not prove an antioxidant or prooxidant effect of the drug on the processes of lipid peroxidation. Apparently, the observed antioxidant effect of rimantadine in vivo is not connected directly with free radical processes in the organism.
Asunto(s)
Antioxidantes/farmacología , Peroxidación de Lípido/efectos de los fármacos , Pulmón/fisiología , Infecciones por Orthomyxoviridae/fisiopatología , Rimantadina/farmacología , Animales , Depuradores de Radicales Libres/farmacología , Virus de la Influenza A , Pulmón/efectos de los fármacos , Pulmón/fisiopatología , Masculino , Ratones , Ratones Endogámicos ICR , Infecciones por Orthomyxoviridae/sangre , Ratas , Ratas Wistar , Superóxidos/metabolismo , Sustancias Reactivas al Ácido Tiobarbitúrico/análisis , Xantina , Xantina OxidasaRESUMEN
The effect of low-intensity laser irradiation on the processes of lipid peroxidation in lens homogenate and aqueous humor during experimental diquat-induced cataract of rabbit eyes was studied. The levels of primary and secondary products of lipid peroxidation (LPO), conjugated dienes and thiobarbituric acid reactive substances (TBARS), were evaluated. We found that the experimental cataract model leads to a significant increase in the content of conjugated dienes and in the content of TBARS both in lens homogenate and in aqueous humor. The data obtained support the important role of oxidative stress in the development of the diquat-induced cataract model. Low-intensity laser treatment does not provoke a significant decrease in conjugated dienes or in TBARS in either lens homogenate or aqueous humor. Although our therapeutic scheme led to a slightly decreased level of LPO products, we conclude that the effect of low-intensity laser-irradiation may depend on the dose applied, individual tissues and other factors.
Asunto(s)
Catarata/metabolismo , Ojo/metabolismo , Rayos Láser/efectos adversos , Peroxidación de Lípido/efectos de la radiación , Animales , Humor Acuoso/metabolismo , Humor Acuoso/efectos de la radiación , Catarata/inducido químicamente , Chinchilla , Diquat , Ojo/efectos de los fármacos , Herbicidas , Cristalino/metabolismo , Cristalino/efectos de la radiación , Masculino , Conejos , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismoRESUMEN
The effect of some psychotropic drugs on the activity of macrophages to produce superoxide radicals during phagocytosis was tested. Three-cyclic antidepressants, imipramine and amitriptyline, and the thioxanthene neuroleptic, chlorprothixene, were studied. The superoxide production was measured by luminol-dependent chemiluminescence. The drugs were investigated in the concentration range of 10(-7)-10(-4) mol/l. It was seen that all tested drugs caused a concentration-dependent decrease of the luminol-dependent chemiluminescence. The inhibitory effect of imipramine and amitriptyline on the macrophage superoxide production was moderate, while the effect of chlorprothixene was significantly stronger (a decrease more than 100 times that of macrophage chemiluminescence). Essentially, the luminol-dependent chemiluminescence reflects the level of superoxide radicals in the system. Therefore, the effect of drugs may be due to the possible activity for scavenging superoxide. In additional experiments with different systems of generations of O2- and different methods of registration, this possibility was discarded. Therefore, the effect of the drugs on the luminol-dependent chemiluminescence seems to be due to drug-induced decrease of the ability of activated macrophages to produce superoxide radicals.
Asunto(s)
Luminol/metabolismo , Macrófagos/efectos de los fármacos , Psicotrópicos/farmacología , Amitriptilina/farmacología , Animales , Clorprotixeno/farmacología , Imipramina/farmacología , Mediciones Luminiscentes , Activación de Macrófagos , Macrófagos/metabolismo , Masculino , Ratas , Ratas Wistar , Superóxidos/metabolismoRESUMEN
The inhibitory effect of some phenothiazine neuroleptics (chlorpromazine, levomepromazine, thioridazine, promethazine and trifluoperazine) on the ability of rat peritoneal macrophages to produce O2- during phagocytosis was investigated. The superoxide radical release was estimated by measuring the luminol-dependent chemiluminescence (CL). The effect of drugs was studied in the concentration range of 0.1-100 mumol/l. Additional experiments to determine the ability of the drugs to scavenge O2- were carried out. They included measuring the effect of phenothiazines on the luminol-dependent CL in systems with enzymatically (xanthine-xanthine oxidase) and non-enzymatically (KO2) generated O2-. The ability of phenothiazines to scavenge O2- was additionally tested by a "non-luminescence" method in which the superoxide concentration was determined spectrophotometrically by the reduction of nitro blue tetrazolium to formazan. All drugs tested decreased significantly CL of stimulated macrophages at concentrations greater than 1 mumol/l. The C50 values were between 0.45 and 1.74 mumol/l. Also phenothiazines were found to act as scavengers of O2-. However, this effect occurred at significantly higher drug concentrations. The C50 values for 50% scavenging of O2- in systems with different sources of O2- were in the concentration range of 5-160 mumol/l. These results suggested that phenothiazines predominantly affected the ability of macrophages to produce O2- during phagocytosis. The findings may provide some insight into the untoward effects of the drugs tested.