RESUMEN
Oxidative stress is induced by accumulation of hydrogen peroxide (H2O2), and therefore, H2O2 could serve as a potential biomarker of various oxidative stress-associated inflammatory diseases. Vanillin is one of the major components of natural vanilla and has potent antioxidant and anti-inflammatory activities. In this work, we developed a novel inflammation-responsive antioxidant polymeric prodrug of vanillin, termed poly(vanillin oxalate) (PVO). In design, PVO incorporates H2O2-reacting peroxalate ester bonds and bioactive vanillin via acid-responsive acetal linkages in its backbone. Therefore, in cells undergoing damages by oxidative stress, PVO readily degrades into three nontoxic components, one of which is antioxidant and anti-inflammatory vanillin. PVO nanoparticles exhibit potent antioxidant activities by scavenging H2O2 and inhibiting the generation of ROS (reactive oxygen species) and also reduce the expression of pro-inflammatory cytokines in activated macrophages in vitro and in vivo. We, therefore, anticipate that PVO nanoparticles have great potential as novel antioxidant therapeutics and drug delivery systems for ROS-associated inflammatory diseases.
Asunto(s)
Antiinflamatorios/síntesis química , Antioxidantes/síntesis química , Benzaldehídos/química , Dioxanos/síntesis química , Peróxido de Hidrógeno/química , Nanopartículas/química , Poliésteres/síntesis química , Profármacos/síntesis química , Animales , Antiinflamatorios/farmacocinética , Antiinflamatorios/farmacología , Antioxidantes/farmacocinética , Antioxidantes/farmacología , Biomarcadores/química , Dioxanos/farmacocinética , Dioxanos/farmacología , Inflamación/prevención & control , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Nanopartículas/uso terapéutico , Estrés Oxidativo , Poliésteres/farmacocinética , Poliésteres/farmacología , Profármacos/farmacocinética , Profármacos/farmacología , Especies Reactivas de Oxígeno/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
The roles that accessory gene products play in activating the Helicobacter pylori urease apoprotein were examined. The activity of the urease apoprotein increased in the following order when it was expressed with the accessory genes: ureGAsunto(s)
Apoproteínas/metabolismo
, Proteínas Bacterianas/metabolismo
, Regulación Bacteriana de la Expresión Génica
, Helicobacter pylori/enzimología
, Ureasa/metabolismo
, Apoproteínas/genética
, Proteínas Bacterianas/genética
, Proteínas Portadoras/genética
, Proteínas Portadoras/metabolismo
, Activación Enzimática
, Immunoblotting
, Proteínas de Unión a Fosfato
, Plásmidos
, Ureasa/genética
RESUMEN
Overproduction of hydrogen peroxide (H2O2) causes oxidative stress and is the main culprit in the pathogenesis of ischemia/reperfusion (I/R) injury. Suppression of oxidative stress is therefore critical in the treatment of I/R injury. Here, we report H2O2-activatable antioxidant prodrug (BRAP) that is capable of specifically targeting the site of oxidative stress and exerting anti-inflammatory and anti-apoptotic activities. BRAP with a self-immolative boronic ester protecting group was designed to scavenge H2O2 and release HBA (p-hydroxybenzyl alcohol) with antioxidant and anti-inflammatory activities. BRAP exerted potent antioxidant and anti-inflammatory activity in lipopolysaccharide (LPS)- and H2O2-stimulated cells by suppressing the generation of ROS and pro-inflammatory cytokines. In mouse models of hepatic I/R and cardiac I/R, BRAP exerted potent antioxidant, anti-inflammatory and anti-apoptotic activities due to the synergistic effects of H2O2-scavenging boronic esters and therapeutic HBA. In addition, administration of high doses of BRAP daily for 7 days showed no renal or hepatic function abnormalities. Therefore BRAP has tremendous therapeutic potential as H2O2-activatable antioxidant prodrug for the treatment of I/R injuries.
Asunto(s)
Antioxidantes/farmacología , Ácidos Borónicos/farmacología , Peróxido de Hidrógeno/antagonistas & inhibidores , Profármacos/farmacología , Daño por Reperfusión/prevención & control , Animales , Antiinflamatorios/farmacología , Antioxidantes/química , Apoptosis/efectos de los fármacos , Alcoholes Bencílicos/farmacología , Ácidos Borónicos/química , Caspasa 3/metabolismo , Línea Celular , Células Cultivadas , Expresión Génica/efectos de los fármacos , Peróxido de Hidrógeno/metabolismo , Immunoblotting , Hígado/irrigación sanguínea , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Ratones Endogámicos BALB C , Microscopía Confocal , Estructura Molecular , Daño por Reperfusión Miocárdica/genética , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/prevención & control , Profármacos/química , Especies Reactivas de Oxígeno/antagonistas & inhibidores , Especies Reactivas de Oxígeno/metabolismo , Daño por Reperfusión/genética , Daño por Reperfusión/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Doxorubicin (DOX) is a commonly used anti-neoplastic agent but its clinical use is limited due to serious hepatic and cardiac side effects. DOX-induced toxicity is mainly associated with overproduction of reactive species oxygen (ROS) such as hydrogen peroxide (H2O2). We have recently developed H2O2-responsive anti-oxidant polymer, polyoxalate containing vanillyl alcohol (PVAX), which is designed to rapidly scavenge H2O2 and release vanillyl alcohol with anti-oxidant, anti-inflammatory and anti-apoptotic properties. In this study, we report that PVAX nanoparticles are novel therapeutic agents for treating DOX-induced cardiac and hepatic toxicity. Intraperitoneal injection of PVAX nanoparticles (4 mg/kg/day) resulted in significant inhibition in apoptosis in liver and heart of DOX-treated mice by suppressing the activation of poly (ADP ribose) polymerase 1 (PARP-1) and caspase-3. PVAX treatment also prevented DOX-induced cardiac dysfunction. Furthermore, survival rate (vehicle = 35% vs. PVAX = 75%; p < 0.05) was significantly improved in a PVAX nanoparticles-treated group compared with vehicle treated groups. Taken together, we anticipate that PVAX nanoparticles could be a highly specific and potent treatment modality in DOX-induced cardiac and hepatic toxicity.
Asunto(s)
Antioxidantes/uso terapéutico , Cardiomiopatías/tratamiento farmacológico , Peróxido de Hidrógeno/metabolismo , Ácido Oxálico/uso terapéutico , Polímeros/uso terapéutico , Animales , Antioxidantes/química , Alcoholes Bencílicos/química , Alcoholes Bencílicos/uso terapéutico , Cardiomiopatías/inducido químicamente , Doxorrubicina , Masculino , Ratones , Ratones Endogámicos C57BL , Nanopartículas/química , Nanopartículas/uso terapéutico , Ácido Oxálico/química , Polímeros/químicaRESUMEN
The main culprit in the pathogenesis of ischemia/reperfusion (I/R) injury is the overproduction of reactive oxygen species (ROS). Hydrogen peroxide (H2O2), the most abundant form of ROS produced during I/R, causes inflammation, apoptosis and subsequent tissue damages. Here, we report H2O2-responsive antioxidant nanoparticles formulated from copolyoxalate containing vanillyl alcohol (VA) (PVAX) as a novel I/R-targeted nanotherapeutic agent. PVAX was designed to incorporate VA and H2O2-responsive peroxalate ester linkages covalently in its backbone. PVAX nanoparticles therefore degrade and release VA, which is able to reduce the generation of ROS, and exert anti-inflammatory and anti-apoptotic activity. In hind-limb I/R and liver I/R models in mice, PVAX nanoparticles specifically reacted with overproduced H2O2 and exerted highly potent anti-inflammatory and anti-apoptotic activities that reduced cellular damages. Therefore, PVAX nanoparticles have tremendous potential as nanotherapeutic agents for I/R injury and H2O2-associated diseases.