RESUMEN
DNA polymerase ß (Pol ß) repairs cellular DNA damage. When such damage is inflicted upon the DNA in tumor cells treated with DNA targeted antitumor agents, Pol ß thus diminishes their efficacy. Accordingly, this enzyme has long been a target for antitumor therapy. Although numerous inhibitors of the lyase activity of the enzyme have been reported, none has yet proven adequate for development as a therapeutic agent. In the present study, we developed a new strategy to identify lyase inhibitors that critically engage the lyase active site primary nucleophile Lys72 as part of the binding interface. This involves a parallel evaluation of the effect of the inhibitors on the wild-type DNA polymerase ß (Pol ß) and Pol ß modified with a lysine analogue at position 72. A model panel of five structurally diverse lyase inhibitors identified in our previous studies (only one of which has been published) with unknown modes of binding were used for testing, and one compound, cis-9,10-epoxyoctadecanoic acid, was found to have the desired characteristics. This finding was further corroborated by in silico docking, demonstrating that the predominant mode of binding of the inhibitor involves an important electrostatic interaction between the oxygen atom of the epoxy group and Nε of the main catalytic nucleophile, Lys72. The strategy, which is designed to identify compounds that engage certain structural elements of the target enzyme, could find broader application for identification of ligands with predetermined sites of binding.
Asunto(s)
ADN Polimerasa beta/metabolismo , Ácidos Esteáricos/metabolismo , Sitios de Unión , Dominio Catalítico , ADN Polimerasa beta/antagonistas & inhibidores , ADN Polimerasa beta/genética , Humanos , Ligandos , Simulación del Acoplamiento Molecular , Mutagénesis Sitio-Dirigida , Unión Proteica , Ácidos Esteáricos/químicaRESUMEN
Friedreich's ataxia (FRDA) is an autosomal recessive neurodegenerative disorder resulting from reduced expression of the protein frataxin (FXN). Although its function is not fully understood, frataxin appears to help assemble iron sulfur clusters; these are critical for the function of many proteins, including those needed for mitochondrial energy production. Finding ways to increase FXN levels has been a major therapeutic strategy for this disease. Previously, we described a novel series of methylene violet analogues and their structural optimization as potential therapeutic agents for neurodegenerative and mitochondrial disorders. Presently, a series of methylene blue analogues has been synthesized and characterized for their in vitro biochemical and biological properties in cultured Friedreich's ataxia lymphocytes. Favorable methylene blue analogues were shown to increase frataxin levels and mitochondrial biogenesis, and to improve aconitase activity. The analogues were found to be good ROS scavengers, and able to protect cultured FRDA lymphocytes from oxidative stress resulting from inhibition of complex I and from glutathione depletion. The analogues also preserved mitochondrial membrane potential and augmented ATP production. Our results suggest that analogue 5, emerging from the initial structure of the parent compound methylene blue (MB), represents a promising lead structure and lacks the cytotoxicity associated with the parent compound MB.
Asunto(s)
Proteínas de Unión a Hierro/metabolismo , Azul de Metileno/análogos & derivados , Mitocondrias/efectos de los fármacos , Adenosina Trifosfato/metabolismo , Línea Celular , Supervivencia Celular/efectos de los fármacos , Complejo I de Transporte de Electrón/metabolismo , Ataxia de Friedreich/metabolismo , Ataxia de Friedreich/patología , Glutatión/metabolismo , Humanos , Linfocitos/citología , Linfocitos/efectos de los fármacos , Linfocitos/metabolismo , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Azul de Metileno/farmacología , Mitocondrias/metabolismo , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , FrataxinaRESUMEN
INTRODUCTION: Our laboratories have demonstrated that accumulation of oligomeric amyloid ß (OAß) in neurons is an essential step leading to OAß-mediated mitochondrial dysfunction. METHODS: Alzheimer's disease (AD) and matching control hippocampal neurons, astrocytes, and microglia were isolated by laser-captured microdissection from the same subjects, followed by whole-transcriptome sequencing. Complementary in vitro work was performed in OAß-treated differentiated SH-SY5Y, followed by the use of a novel CoQ10 analogue for protection. This compound is believed to be effective both in suppressing reactive oxygen species and also functioning in mitochondrial electron transport. RESULTS: We report decreases in the same mitochondrial-encoded mRNAs in Alzheimer's disease laser-captured CA1 neurons and in OAß-treated SH-SY5Y cells, but not in laser-captured microglia and astrocytes. Pretreatment with a novel CoQ10 analogue, protects neuronal mitochondria from OAß-induced mitochondrial changes. DISCUSSION: Similarity of expression changes in neurons from Alzheimer's disease brain and neuronal cells treated with OAß, and the effect of a CoQ10 analogue on the latter, suggests a pretreatment option to prevent OAß toxicity, long before the damage is apparent.
Asunto(s)
Péptidos beta-Amiloides/metabolismo , Neuronas/metabolismo , ARN Mensajero/metabolismo , ARN Mitocondrial/metabolismo , Anciano , Enfermedad de Alzheimer/metabolismo , Astrocitos/efectos de los fármacos , Astrocitos/metabolismo , Línea Celular Tumoral , Femenino , Hipocampo/metabolismo , Humanos , Técnicas In Vitro , Captura por Microdisección con Láser , Masculino , Microglía/efectos de los fármacos , Microglía/metabolismo , Microscopía Electrónica de Transmisión , Neuronas/efectos de los fármacos , ARN Mensajero/genética , ARN Mitocondrial/genética , Ubiquinona/análogos & derivados , Ubiquinona/farmacologíaRESUMEN
In an effort to identify methylene blue analogues having improved antioxidant activity, a series of new methylene violet analogues have been designed and synthesized. The analogues were prepared following a synthetic route that is more efficient than the previously reported methods, both in terms of yield and purity of the final products. The route involves the Smiles rearrangement as one of the crucial steps. Smiles rearrangement of suitably substituted diphenyl sulfide intermediates afforded the corresponding phenothiazine analogues in high yields, which were subsequently converted to the final products. The methylene violet analogues were evaluated for their ability to preserve mitochondrial function in Friedreich's ataxia (FRDA) lymphocytes. The analogues were shown to be efficient ROS scavengers, and able to protect cultured FRDA lymphocytes from oxidative stress resulting from inhibition of complex I. The analogues also preserved mitochondrial membrane potential and augmented ATP production. The analogues were found to be better antioxidants than the parent compounds methylene blue and methylene violet.
Asunto(s)
Mitocondrias/efectos de los fármacos , Fenotiazinas/farmacología , Adenosina Trifosfato/biosíntesis , Células Cultivadas , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Linfocitos/efectos de los fármacos , Linfocitos/metabolismo , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Mitocondrias/metabolismo , Estrés Oxidativo/efectos de los fármacos , Fenotiazinas/síntesis química , Fenotiazinas/química , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Recently, we described the optimization of novel pyrimidinol-based antioxidants as potential therapeutic molecules for targeting mitochondrial diseases. That study focused on improving the potency and metabolic stability of pyrimidinol antioxidants. This led us to consider the possibility of altering the positions of the exocyclic alkoxy and alkylamino substituents on the pyrimidinol scaffold. Twelve new analogues were prepared and their biological activities were investigated. The metabolic stability of the prepared regioisomers was also assessed in vitro using bovine liver microsomes. Unexpectedly, the 2-alkoxy-4-alkylamino substituted pyrimidinol antioxidants were found to have properties in protecting mitochondrial function superior to the isomeric 4-alkoxy-2-alkylamino substituted pyrimidinols evaluated in all earlier studies. This observation suggests a possible mode of action involving the intermediacy of an ortho-iminoquinone, a species not previously associated with mitochondrial respiratory chain function.
Asunto(s)
Antioxidantes/farmacología , Citoprotección/efectos de los fármacos , Pirimidinas/farmacología , Adenosina Trifosfato/metabolismo , Animales , Espectroscopía de Resonancia Magnética con Carbono-13 , Bovinos , Espectrometría de Masas , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Espectroscopía de Protones por Resonancia MagnéticaRESUMEN
In a recent study, several new derivatives of antimycin A (AMA) were produced by means of a novel transacylation reaction, and these were shown to mediate selective toxicity toward cultured A549 human lung epithelial adenocarcinoma cells, as compared with WI-38 normal human lung fibroblasts. The purpose of our study was to investigate whether the analogues all expressed their cytotoxicity by the same mechanism. This was done by studying the effects of the compounds in several types of cell lines. In comparison with 2-O-methylantimycin, which acts at the locus of Bcl-2, none of the new derivatives exhibited a difference in cytotoxicity toward cells expressing different levels of Bcl-2. In cell lines that over- or underexpress estrogen or Her2 receptors, AMA analogue 2 exhibited Her2 receptor dependency at low concentration. Three compounds (1, 4, and 6) exhibited concentration-dependent increases in reactive oxygen species, with 6 being especially potent. Compounds 5 and 6 diminished mitochondrial membrane potential more potently than AMA, and 1 also displayed enhanced activity relative to 2-4. Interestingly, only 1 and AMA displayed strong inhibition of the respiratory chain, as measured by monitoring NADH (reduced nicotinamide adenine dinucleotide) oxidase. Because four of the analogues have positively charged substituents, two of these (4 and 6) were studied to see whether the observed effects were due to much higher level of accumulation within the mitochondria. Their presence in the mitochondria was not dramatically enhanced. Neither of the two presently characterized mechanisms of cell killing by AMA can fully account for the observed results.
Asunto(s)
Antimicina A/análogos & derivados , Citotoxinas/farmacología , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Complejos Multienzimáticos/antagonistas & inhibidores , NADH NADPH Oxidorreductasas/antagonistas & inhibidores , Especies Reactivas de Oxígeno/metabolismo , Acilación , Animales , Antimicina A/química , Ácido Ascórbico/análogos & derivados , Ácido Ascórbico/farmacología , Bovinos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Citotoxinas/química , Fibroblastos/efectos de los fármacos , Humanos , Concentración 50 Inhibidora , Proteínas Proto-Oncogénicas c-bcl-2/efectos de los fármacos , Proteínas Proto-Oncogénicas c-bcl-2/genéticaRESUMEN
INTRODUCTION: We have comprehensively described the expression profiles of mitochondrial DNA and nuclear DNA genes that encode subunits of the respiratory oxidative phosphorylation (OXPHOS) complexes (I-V) in the hippocampus from young controls, age matched, mild cognitively impaired (MCI), and Alzheimer's disease (AD) subjects. METHODS: Hippocampal tissues from 44 non-AD controls (NC), 10 amnestic MCI, and 18 AD cases were analyzed on Affymetrix Hg-U133 plus 2.0 arrays. RESULTS: The microarray data revealed significant down regulation in OXPHOS genes in AD, particularly those encoded in the nucleus. In contrast, there was up regulation of the same gene(s) in MCI subjects compared to AD and ND cases. No significant differences were observed in mtDNA genes identified in the array between AD, ND, and MCI subjects except one mt-ND6. DISCUSSION: Our findings suggest that restoration of the expression of nuclear-encoded OXPHOS genes in aging could be a viable strategy for blunting AD progression.
Asunto(s)
Envejecimiento/genética , Enfermedad de Alzheimer/genética , Trastornos del Conocimiento/genética , Mitocondrias/genética , Fosforilación Oxidativa , Adulto , Anciano de 80 o más Años , Autopsia , Femenino , Hipocampo , Humanos , Masculino , Análisis de Secuencia por Matrices de OligonucleótidosRESUMEN
Nitroreductase (NTR) activities have been known for decades, studied extensively in bacteria and also in systems as diverse as yeast, trypanosomes, and hypoxic tumors. The putative bacterial origin of mitochondria prompted us to explore the possible existence of NTR activity within this organelle and to probe its behavior in a cellular context. Presently, by using a profluorescent near-infrared (NIR) dye, we characterize the nature of NTR activity localized in mammalian cell mitochondria. Further, we demonstrate that this mitochondrially localized enzymatic activity can be exploited both for selective NIR imaging of mitochondria and for mitochondrial targeting by activating a mitochondrial poison specifically within that organelle. This constitutes a new mechanism for mitochondrial imaging and targeting. These findings represent the first use of mitochondrial enzyme activity to unmask agents for mitochondrial fluorescent imaging and therapy, which may prove to be more broadly applicable.
Asunto(s)
Mitocondrias/enzimología , Nitrorreductasas/metabolismo , Células A549 , Antimicina A/análogos & derivados , Antimicina A/química , Antimicina A/farmacología , Escherichia coli/enzimología , Colorantes Fluorescentes/química , Humanos , Mitocondrias/efectos de los fármacos , Estructura Molecular , Nitrorreductasas/genética , Imagen Óptica , Espectroscopía Infrarroja CortaRESUMEN
Previously we described a novel series of pyrimidinol antioxidants and their structural optimization as potential therapeutic agents for neurodegenerative and mitochondrial disorders. Our initial lead compound was a potent antioxidant in vitro, but was subsequently found to exhibit poor stability to oxidative metabolism. The current study focused on balancing potency with metabolic stability through structural modification, and involved modifications at positions 2 and 4 of the pyrimidinol redox core, likely sites of oxidative metabolism. Eight new analogues have been prepared and their ability to suppress lipid peroxidation and reactive oxygen species (ROS), and to preserve mitochondrial membrane potential (Δψm) and support ATP production, has been investigated. The metabolic stability of the prepared compounds was also assessed in vitro using bovine liver microsomes to obtain preliminary insight on this class of compounds. This study revealed the complexity of balancing reasonable metabolic stability with efficient antioxidant properties. While a few analogues appear promising, especially in terms of metabolic stability, a 4-isopropoxy derivative conserved the favorable biological activity and exhibited good metabolic stability. The favorable metabolic stability conferred by the combination of the azetidine and isopropoxy moieties in analogue 6 makes this compound an excellent candidate for further evaluation.
Asunto(s)
Adenosina Trifosfato/biosíntesis , Antioxidantes/farmacología , Microsomas Hepáticos/química , Mitocondrias/efectos de los fármacos , Sustancias Protectoras/farmacología , Pirimidinas/farmacología , Animales , Antioxidantes/química , Antioxidantes/metabolismo , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Humanos , Peroxidación de Lípido/efectos de los fármacos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Estructura Molecular , Sustancias Protectoras/química , Sustancias Protectoras/metabolismo , Pirimidinas/química , Pirimidinas/metabolismo , Especies Reactivas de Oxígeno/antagonistas & inhibidores , Especies Reactivas de Oxígeno/metabolismo , Relación Estructura-Actividad , PorcinosRESUMEN
As part of our ongoing efforts to identify compounds having potential utility in treating neurodegenerative and mitochondrial disorders, a series of pyridinol analogues have been prepared. The synthetic route employed for the preparation of the new analogues is different, and considerably more efficient, than that used in previously reported studies. The new route yields a pair of pyridinol regioisomers that can be readily separated and evaluated. Their ability to quench lipid peroxidation and reactive oxygen species (ROS), and to preserve mitochondrial membrane potential (Δψm) and support ATP synthesis is reported. The optimal side chain length was found to be 16 carbon atoms. The metabolic stability of those compounds having optimal biological activities was evaluated in vitro using bovine liver microsomes. The omission of any side chain hydroxyl group and introduction of an azetidine moiety at position 6 of the pyridinol redox core (8 and 9) increased their microsomal stability as compared to the exocyclic dimethylamino group. The favorable metabolic stability conferred by the azetidine moiety in compounds 8 and 9 makes these compounds excellent candidates for further evaluation.
Asunto(s)
Antioxidantes/farmacología , Citoprotección/efectos de los fármacos , Enfermedades Mitocondriales/tratamiento farmacológico , Enfermedades Neurodegenerativas/tratamiento farmacológico , Fármacos Neuroprotectores/farmacología , Piridinas/farmacología , Animales , Antioxidantes/síntesis química , Antioxidantes/química , Bovinos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Humanos , Peroxidación de Lípido/efectos de los fármacos , Linfocitos/efectos de los fármacos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Microsomas Hepáticos/química , Microsomas Hepáticos/metabolismo , Enfermedades Mitocondriales/patología , Estructura Molecular , Enfermedades Neurodegenerativas/patología , Fármacos Neuroprotectores/síntesis química , Fármacos Neuroprotectores/química , Piridinas/síntesis química , Piridinas/química , Especies Reactivas de Oxígeno/antagonistas & inhibidores , Especies Reactivas de Oxígeno/metabolismo , Relación Estructura-ActividadRESUMEN
Two new aza analogues of the neuroprotective agent idebenone have been synthesized and characterized. Their antioxidant activity, and ability to augment ATP levels have been evaluated in several different cell lines having suboptimal mitochondrial function. Both compounds were found to be good ROS scavengers, and to protect the cells from oxidative stress induced by glutathione depletion. The compounds were more effective than idebenone in neurodegenerative disease cells. These novel pyrimidinol derivatives were also shown to augment ATP levels in coenzyme Q(10)-deficient human lymphocytes. The more lipophilic side chains attached to the pyrimidinol redox core in these compounds resulted in less inhibition of the electron transport chain and improved antioxidant activity.
Asunto(s)
Antioxidantes/química , Mitocondrias/metabolismo , Fármacos Neuroprotectores/química , Pirimidinas/química , Ubiquinona/análogos & derivados , Adenosina Trifosfato/metabolismo , Animales , Antioxidantes/síntesis química , Antioxidantes/toxicidad , Bovinos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Diseño de Fármacos , Glutatión/metabolismo , Humanos , Linfocitos/efectos de los fármacos , Linfocitos/metabolismo , Mitocondrias/enzimología , Complejos Multienzimáticos/antagonistas & inhibidores , Complejos Multienzimáticos/metabolismo , NADH NADPH Oxidorreductasas/antagonistas & inhibidores , NADH NADPH Oxidorreductasas/metabolismo , Enfermedades Neurodegenerativas/metabolismo , Enfermedades Neurodegenerativas/patología , Fármacos Neuroprotectores/síntesis química , Fármacos Neuroprotectores/toxicidad , Pirimidinas/síntesis química , Pirimidinas/toxicidad , Especies Reactivas de Oxígeno/metabolismo , Ubiquinona/síntesis química , Ubiquinona/química , Ubiquinona/genética , Ubiquinona/metabolismo , Ubiquinona/toxicidadRESUMEN
The effect of the alkyl side chain length of coenzyme Q10 on mitochondrial respiratory chain function has been investigated by the use of synthetic ubiquinone derivatives. Three analogues (3, 4 and 6) were identified that exhibited significantly improved effects on mitochondrial oxygen consumption and mitochondrial membrane potential, and also conferred significant cytoprotection on cultured mammalian cells in which glutathione had been depleted by treatment with diethyl maleate. The analogues also exhibited lesser inhibition of the electron transport chain than idebenone. The results obtained provide guidance for the design of CoQ10 analogues with improved activity compared to that of idebenone (1), the latter of which is undergoing evaluation in the clinic as a therapeutic agent.
Asunto(s)
Transporte de Electrón/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Ubiquinona/análogos & derivados , Animales , Bovinos , Línea Celular , Línea Celular Tumoral , Citoprotección , Transporte de Electrón/fisiología , Humanos , Leucemia/metabolismo , Leucemia/patología , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Mitocondrias/metabolismo , Consumo de Oxígeno/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Células Ganglionares de la Retina/citología , Células Ganglionares de la Retina/efectos de los fármacos , Células Ganglionares de la Retina/metabolismo , Relación Estructura-Actividad , Ubiquinona/química , Ubiquinona/metabolismo , Ubiquinona/farmacologíaRESUMEN
Bicyclic pyridinol antioxidants have been reported to suppress the autoxidation of methyl linoleate more effectively than α-tocopherol in benzene solution. A few novel lipophilic analogues have recently been synthesized by conjugating a pyridinol core with the phytyl side chain of α-tocopherol; these have been shown to possess potent antioxidant activity. However, the complexity of the synthetic routes has hampered their further development. Herein, we describe a facile approach, involving only five synthetic steps, to simplified analogues (1a-1c) and their acetate ester precursors (2a-2c). Simple alkyl chains of different lengths were attached to the 6-methyl group of the antioxidant core via regioselective metalation. These analogues were found to retain biological activity and exhibit protective behaviour under conditions of induced oxidative stress, which could lead to the development of more readily accessible analogues as potential antioxidants capable of preserving mitochondrial function.
Asunto(s)
Antioxidantes/química , Antioxidantes/farmacología , Mitocondrias/efectos de los fármacos , Piridinas/síntesis química , Piridinas/farmacología , Antioxidantes/síntesis química , Línea Celular Tumoral , Diseño de Fármacos , Evaluación Preclínica de Medicamentos , Humanos , Leucemia/tratamiento farmacológico , Peroxidación de Lípido/efectos de los fármacos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Mitocondrias/metabolismo , Fenómenos Químicos Orgánicos , Estrés Oxidativo/efectos de los fármacos , Piridinas/química , Piridonas , Especies Reactivas de Oxígeno/metabolismo , Relación Estructura-Actividad , alfa-Tocoferol/análogos & derivadosRESUMEN
Selected pyridinol analogues of the experimental neuroprotective drug idebenone have been synthesized and evaluated as antioxidants capable of preserving mitochondrial function. The compounds, having a different redox core but the same side chain as idebenone, exhibited a range of potencies, reflecting differences in their structures. The results obtained provide guidance in the design of such analogues with improved properties. Analogues were identified that have significantly improved antioxidant activity compared with idebenone in cultured lymphocytes, and which exhibit lesser inhibition of the electron transport chain.
Asunto(s)
Antioxidantes/farmacología , Citoprotección/efectos de los fármacos , Mitocondrias Cardíacas/efectos de los fármacos , Piridonas/farmacología , Animales , Antioxidantes/síntesis química , Antioxidantes/química , Bovinos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Humanos , Peroxidación de Lípido/efectos de los fármacos , Linfocitos/efectos de los fármacos , Linfocitos/metabolismo , Mitocondrias Cardíacas/enzimología , Mitocondrias Cardíacas/metabolismo , Membranas Mitocondriales/efectos de los fármacos , Membranas Mitocondriales/metabolismo , Estructura Molecular , Complejos Multienzimáticos/antagonistas & inhibidores , Complejos Multienzimáticos/metabolismo , NADH NADPH Oxidorreductasas/antagonistas & inhibidores , NADH NADPH Oxidorreductasas/metabolismo , Piridonas/síntesis química , Piridonas/química , Especies Reactivas de Oxígeno/antagonistas & inhibidores , Especies Reactivas de Oxígeno/metabolismo , Relación Estructura-Actividad , Células Tumorales CultivadasRESUMEN
The synthesis of benzoquinone natural product 2 and three analogues is described. The synthesized compounds were tested for their ability to protect Friedreich's ataxia (FRDA) lymphocytes from induced oxidative stress. One of the analogues (3) conferred cytoprotection in a dose-dependent manner in FRDA lymphocytes at micromolar concentrations. The biological assays suggest that the modification of the 2-hydroxy and N-(3-carboxypropyl) groups in the natural product can improve its antioxidant activity and significantly enhance its ability to protect mitochondrial function under conditions of oxidative stress.
Asunto(s)
Benzoquinonas/química , Benzoquinonas/farmacología , Ácido gamma-Aminobutírico/análogos & derivados , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ataxia de Friedreich/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Estructura Molecular , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/farmacología , Relación Estructura-Actividad , Ácido gamma-Aminobutírico/química , Ácido gamma-Aminobutírico/farmacologíaRESUMEN
PURPOSE: To investigate of an approach to stabilize a novel pyridinol based α-tocopherol analogue (1) as a prodrug by acetylation of its phenol moiety. METHODS: Biochemical indicators of oxidative stress in mitochondria were utilized to gain insight into the cytoprotective mechanism(s) of compound 1 acetate. Oxygen free radical scavenging activity was measured using DCF probe in a cultured cell model system that had been placed under oxidative stress. Lipid peroxidation was examined both in a cell-free system and in oxidatively stressed cultured cells. The bioenergetic parameters of mitochondria were evaluated by measuring mitochondrial membrane potential (Δψ(m)) and the MPT. RESULTS: The present results suggest strongly that the antioxidant efficacy of compound 1 can be improved by using it as a prodrug. The tested prodrug has shown to be activated as a function of time, presumably due to susceptibility to enzymatic hydrolysis, and exhibits an antioxidant effect in time-dependent manner, providing a compound that is more effective than α-tocopherol acetate with regard to all protective properties studied. CONCLUSIONS: An effective approach to stabilize compound 1 was realized by using its acetate as a prodrug.
Asunto(s)
Antioxidantes/química , Sistemas de Liberación de Medicamentos , Liposomas/síntesis química , Profármacos/síntesis química , Vitamina E/análogos & derivados , Vitamina E/química , Acetatos/análisis , Acetatos/síntesis química , Acetatos/química , Acetatos/metabolismo , Antioxidantes/metabolismo , Antioxidantes/farmacología , Compuestos de Boro/metabolismo , Técnicas de Cultivo de Célula , Línea Celular Tumoral , Supervivencia Celular , Sistema Libre de Células , Composición de Medicamentos , Evaluación Preclínica de Medicamentos , Humanos , Peroxidación de Lípido , Liposomas/química , Potencial de la Membrana Mitocondrial , Mitocondrias/metabolismo , Oxidación-Reducción , Estrés Oxidativo , Profármacos/química , Profármacos/metabolismo , Profármacos/farmacología , Especies Reactivas de Oxígeno/análisis , Especies Reactivas de Oxígeno/metabolismo , Vitamina E/metabolismo , Vitamina E/farmacologíaRESUMEN
An efficient synthesis has provided access to a novel α-tocopherol analogue (2), as well as its trifluoroacetate salt and acetate ester. An annulation reaction was used to establish the pyridinol core structure and a Stille coupling reaction was employed for conjugation with the tocopherol side chain. This analogue was shown to suppress the levels of reactive oxygen species in cultured cells, and to quench peroxidation of mitochondrial membranes.
Asunto(s)
Antioxidantes/química , Diterpenos/síntesis química , Mitocondrias/metabolismo , Piridinas/síntesis química , alfa-Tocoferol/análogos & derivados , Animales , Antioxidantes/metabolismo , Bovinos , Membrana Celular/metabolismo , Diterpenos/química , Diterpenos/farmacología , Diseño de Fármacos , Peroxidación de Lípido/efectos de los fármacos , Piridinas/química , Piridinas/farmacología , Pirimidinas/química , Pirroles/química , Especies Reactivas de Oxígeno/metabolismo , alfa-Tocoferol/síntesis química , alfa-Tocoferol/farmacologíaRESUMEN
Verticipyrone has recently been isolated from the culture broth of Verticillium sp. and shown to inhibit NADH fumarate reductase, as well as NADH oxidoreductase (complex I) of the mitochondrial electron transport chain. In order to assess the structural elements in verticipyrone essential for complex I inhibitor, 15 structural analogues were prepared and analyzed for their effects on mitochondrial NADH oxidoreductase and NADH oxidase activities. Also measured were the abilities of several of the analogues to inhibit respiration as judged by a shift to glycolysis, and to inhibit the growth of several mammalian cell lines. The nature of the pyrone ring was shown to be important to potency of inhibition, as was the length and nature of substituents in the side chain of the analogues.
Asunto(s)
Alquenos/farmacología , Mitocondrias/efectos de los fármacos , Pironas/farmacología , Relación Estructura-Actividad , Alquenos/química , Mitocondrias/fisiología , Pironas/químicaRESUMEN
Ferroptosis is an iron-catalyzed, nonapoptotic form of regulated necrosis that has been implicated in the pathological cell death associated with various disorders including neurodegenerative diseases (e.g., Friedreich's ataxia (FRDA), Alzheimer's disease, and Parkinson's disease), stroke, and traumatic brain injury. Recently, we showed that lipophilic methylene blue (MB) and methylene violet (MV) analogues both promoted increased frataxin levels and mitochondrial biogenesis, in addition to their antioxidant activity in cultured FRDA cells. Presently, we report the synthesis of series of lipophilic phenothiazine analogues that potently inhibit ferroptosis. The most promising compounds (1b-5b) exhibited an improved protection compared to the parent phenothiazine against erastin- and RSL3-induced ferroptotic cell death. These analogues have equivalent or better potency than ferrostatin-1 (Fer-1) and liproxstatin-1 (Lip-1), that are among the most potent inhibitors of this regulated cell death described so far. They represent novel lead compounds with therapeutic potential in relevant ferroptosis-driven disease models such as FRDA.
RESUMEN
There has been little innovation in identifying novel insulin sensitizers. Metformin, developed in the 1920s, is still used first for most Type 2 diabetes patients. Mice with genetic reduction of p52Shc protein have improved insulin sensitivity and glucose tolerance. By high-throughput screening, idebenone was isolated as the first small molecule 'Shc Blocker'. Idebenone blocks p52Shc's access to Insulin Receptor to increase insulin sensitivity. In this work the avidity of 34 novel idebenone analogs and 3 metabolites to bind p52Shc, and to block the interaction of p52Shc with the Insulin receptor was tested. Our hypothesis was that if an idebenone analog bound and blocked p52Shc's access to insulin receptor better than idebenone, it should be a more effective insulin sensitizing agent than idebenone itself. Of 34 analogs tested, only 2 both bound p52Shc more tightly and/or blocked the p52Shc-Insulin Receptor interaction more effectively than idebenone. Of those 2 only idebenone analog #11 was a superior insulin sensitizer to idebenone. Also, the long-lasting insulin-sensitizing potency of idebenone in rodents over many hours had been puzzling, as the parent molecule degrades to metabolites within 1 h. We observed that two of the idebenone's three metabolites are insulin sensitizing almost as potently as idebenone itself, explaining the persistent insulin sensitization of this rapidly metabolized molecule. These results help to identify key SAR = structure-activity relationship requirements for more potent small molecule Shc inhibitors as Shc-targeted insulin sensitizers for type 2 diabetes.