ABSTRACT
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.
Subject(s)
Antioxidants/pharmacology , Boronic Acids/pharmacology , Hydrogen Peroxide/antagonists & inhibitors , Prodrugs/pharmacology , Reperfusion Injury/prevention & control , Animals , Anti-Inflammatory Agents/pharmacology , Antioxidants/chemistry , Apoptosis/drug effects , Benzyl Alcohols/pharmacology , Boronic Acids/chemistry , Caspase 3/metabolism , Cell Line , Cells, Cultured , Gene Expression/drug effects , Hydrogen Peroxide/metabolism , Immunoblotting , Liver/blood supply , Liver/drug effects , Liver/metabolism , Male , Mice, Inbred BALB C , Microscopy, Confocal , Molecular Structure , Myocardial Reperfusion Injury/genetics , Myocardial Reperfusion Injury/metabolism , Myocardial Reperfusion Injury/prevention & control , Prodrugs/chemistry , Reactive Oxygen Species/antagonists & inhibitors , Reactive Oxygen Species/metabolism , Reperfusion Injury/genetics , Reperfusion Injury/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolismABSTRACT
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.
Subject(s)
Antioxidants/therapeutic use , Cardiomyopathies/drug therapy , Hydrogen Peroxide/metabolism , Oxalic Acid/therapeutic use , Polymers/therapeutic use , Animals , Antioxidants/chemistry , Benzyl Alcohols/chemistry , Benzyl Alcohols/therapeutic use , Cardiomyopathies/chemically induced , Doxorubicin , Male , Mice , Mice, Inbred C57BL , Nanoparticles/chemistry , Nanoparticles/therapeutic use , Oxalic Acid/chemistry , Polymers/chemistryABSTRACT
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.
Subject(s)
Antioxidants/metabolism , Hydrogen Peroxide/metabolism , Nanoparticles/metabolism , Polymers/metabolism , Reperfusion Injury/metabolism , Animals , Anti-Inflammatory Agents/administration & dosage , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/metabolism , Antioxidants/administration & dosage , Antioxidants/chemistry , Apoptosis , Benzyl Alcohols/chemistry , Cell Line , Inflammation/drug therapy , Inflammation/metabolism , Male , Mice , Nanoparticles/administration & dosage , Nanoparticles/chemistry , Nanoparticles/ultrastructure , Polymers/administration & dosage , Polymers/chemistry , Reactive Oxygen Species/metabolism , Reperfusion Injury/drug therapyABSTRACT
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.
Subject(s)
Anti-Inflammatory Agents/chemical synthesis , Antioxidants/chemical synthesis , Benzaldehydes/chemistry , Dioxanes/chemical synthesis , Hydrogen Peroxide/chemistry , Nanoparticles/chemistry , Polyesters/chemical synthesis , Prodrugs/chemical synthesis , Animals , Anti-Inflammatory Agents/pharmacokinetics , Anti-Inflammatory Agents/pharmacology , Antioxidants/pharmacokinetics , Antioxidants/pharmacology , Biomarkers/chemistry , Dioxanes/pharmacokinetics , Dioxanes/pharmacology , Inflammation/prevention & control , Macrophages/drug effects , Macrophages/metabolism , Mice , Nanoparticles/therapeutic use , Oxidative Stress , Polyesters/pharmacokinetics , Polyesters/pharmacology , Prodrugs/pharmacokinetics , Prodrugs/pharmacology , Reactive Oxygen Species/antagonists & inhibitors , Tumor Necrosis Factor-alpha/antagonists & inhibitors , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
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: ureGSubject(s)
Apoproteins/metabolism
, Bacterial Proteins/metabolism
, Gene Expression Regulation, Bacterial
, Helicobacter pylori/enzymology
, Urease/metabolism
, Apoproteins/genetics
, Bacterial Proteins/genetics
, Carrier Proteins/genetics
, Carrier Proteins/metabolism
, Enzyme Activation
, Immunoblotting
, Phosphate-Binding Proteins
, Plasmids
, Urease/genetics