RESUMO
BACKGROUND: Photodynamic therapy (PDT) is a promising therapeutic modality that can convert oxygen into cytotoxic reactive oxygen species (ROS) via photosensitizers to halt tumor growth. However, hypoxia and the unsatisfactory accumulation of photosensitizers in tumors severely diminish the therapeutic effect of PDT. In this study, a multistage nanoplatform is demonstrated to overcome these limitations by encapsulating photosensitizer IR780 and oxygen regulator 3-bromopyruvate (3BP) in poly (lactic-co-glycolic acid) (PLGA) nanocarriers. RESULTS: The as-synthesized nanoplatforms penetrated deeply into the interior region of tumors and preferentially remained in mitochondria due to the intrinsic characteristics of IR780. Meanwhile, 3BP could efficiently suppress oxygen consumption of tumor cells by inhibiting mitochondrial respiratory chain to further improve the generation of ROS. Furthermore, 3BP could abolish the excessive glycolytic capacity of tumor cells and lead to the collapse of ATP production, rendering tumor cells more susceptible to PDT. Successful tumor inhibition in animal models confirmed the therapeutic precision and efficiency. In addition, these nanoplatforms could act as fluorescence (FL) and photoacoustic (PA) imaging contrast agents, effectuating imaging-guided cancer treatment. CONCLUSIONS: This study provides an ideal strategy for cancer therapy by concurrent oxygen consumption reduction, oxygen-augmented PDT, energy supply reduction, mitochondria-targeted/deep-penetrated nanoplatforms and PA/FL dual-modal imaging guidance/monitoring. It is expected that such strategy will provide a promising alternative to maximize the performance of PDT in preclinical/clinical cancer treatment.
Assuntos
Mitocôndrias/efeitos dos fármacos , Nanopartículas/química , Fármacos Fotossensibilizantes/farmacologia , Hipóxia Tumoral/efeitos dos fármacos , Animais , Linhagem Celular Tumoral , Sinergismo Farmacológico , Feminino , Humanos , Indóis/química , Indóis/farmacocinética , Indóis/farmacologia , Indóis/uso terapêutico , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Camundongos Nus , Mitocôndrias/metabolismo , Neoplasias/tratamento farmacológico , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/farmacocinética , Fármacos Fotossensibilizantes/uso terapêutico , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Piruvatos/química , Piruvatos/farmacocinética , Piruvatos/farmacologia , Piruvatos/uso terapêutico , Espécies Reativas de Oxigênio/metabolismo , Distribuição Tecidual , Transplante HeterólogoRESUMO
Haloacids are considered to be environmental pollutants, but some of them have also been tested in clinical research. The way that haloacids are transported across biological membranes is important for both biodegradation and drug delivery purposes. In this review, we will first summarize putative haloacids transporters and the information about haloacids transport when studying carboxylates transporters. We will then introduce MCT1 and SLC5A8, which are respective transporter for antitumor agent 3-bromopyruvic acid and dichloroacetic acid, and monochloroacetic acid transporters Deh4p and Dehp2 from a haloacids-degrading bacterium. Phylogenetic analysis of these haloacids transporters and other monocarboxylate transporters reveals their evolutionary relationships. Haloacids transporters are not studied to the extent that they deserve compared with their great application potentials, thus future inter-discipline research are desired to better characterize their transport mechanisms for potential applications in both environmental and clinical fields.
Assuntos
Ácido Dicloroacético/farmacocinética , Piruvatos/farmacocinética , Animais , Transporte Biológico , Membrana Celular/metabolismo , Humanos , Transportadores de Ácidos Monocarboxílicos/fisiologia , Filogenia , Simportadores/fisiologiaRESUMO
PURPOSE: To investigate whether inhalation of ethyl pyruvate (EP) encapsulated with poly(ethylene glycol)-block-lactide/glycolide copolymer nanoparticles (EP-NPs) can prevent the development of shunt-flow-induced hyperkinetic pulmonary arterial hypertension (PAH) in a rat model. MATERIALS AND METHODS: Rats were separated into five groups: blank (ie, no treatment after shunt flow), normal control (ie, no shunt flow or treatment), EP-NP instillation, EP-only instillation, and vehicle. The animals received intratracheal instillation of EP-NPs or other treatments immediately after a shunt flow, and treatment continued weekly until the end of the experiment. Hemodynamic data were recorded, pulmonary arterial remodeling was assessed, and levels of inflammatory mediators and ET1 expression in the lung and serum were analyzed. In addition, retention of EP in the lungs of rats in the EP-NP and EP-only groups was measured using high-performance liquid chromatography. RESULTS: After 12 weeks, hemodynamic abnormalities and pulmonary arterial remodeling were improved in the EP-NP instillation group, compared with the blank, EP-only, and vehicle groups (P<0.05). In addition, the EP-NP group showed significantly decreased levels of HMGB1, IL-6, TNFα, reactive oxygen species, and ET1 in the lung during PAH development (P<0.05). Furthermore, EP-NP instillation was associated with reduced serum levels of inflammatory factors and ET1. High-performance liquid-chromatography measurement indicated that EP retention was greater in the lungs of the EP-NP group than in the EP-only group. CONCLUSION: EP-NP instillation attenuated inflammation and prevented pulmonary arterial remodeling during the development of PAH induced by shunt flow. In the future, EP-NP delivery into the lung might provide a novel approach for preventing PAH.
Assuntos
Hipertensão Pulmonar/prevenção & controle , Nanopartículas/administração & dosagem , Piruvatos/administração & dosagem , Animais , Modelos Animais de Doenças , Endotelina-1/metabolismo , Hipertensão Pulmonar/etiologia , Interleucina-6/metabolismo , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Masculino , Nanopartículas/química , Poliésteres/química , Polietilenoglicóis/química , Substâncias Protetoras/administração & dosagem , Substâncias Protetoras/química , Artéria Pulmonar/efeitos dos fármacos , Artéria Pulmonar/fisiopatologia , Piruvatos/química , Piruvatos/farmacocinética , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/metabolismo , Remodelação Vascular/efeitos dos fármacos , Remodelação Vascular/fisiologiaRESUMO
Although the anti-cancer properties of 3BP (3-bromopyruvate) have been described previously, its selectivity for cancer cells still needs to be explained [Ko et al. (2001) Cancer Lett. 173, 83-91]. In the present study, we characterized the kinetic parameters of radiolabelled [14C] 3BP uptake in three breast cancer cell lines that display different levels of resistance to 3BP: ZR-75-1 < MCF-7 < SK-BR-3. At pH 6.0, the affinity of cancer cells for 3BP transport correlates with their sensitivity, a pattern that does not occur at pH 7.4. In the three cell lines, the uptake of 3BP is dependent on the protonmotive force and is decreased by MCTs (monocarboxylate transporters) inhibitors. In the SK-BR-3 cell line, a sodium-dependent transport also occurs. Butyrate promotes the localization of MCT-1 at the plasma membrane and increases the level of MCT-4 expression, leading to a higher sensitivity for 3BP. In the present study, we demonstrate that this phenotype is accompanied by an increase in affinity for 3BP uptake. Our results confirm the role of MCTs, especially MCT-1, in 3BP uptake and the importance of cluster of differentiation (CD) 147 glycosylation in this process. We find that the affinity for 3BP transport is higher when the extracellular milieu is acidic. This is a typical phenotype of tumour microenvironment and explains the lack of secondary effects of 3BP already described in in vivo studies [Ko et al. (2004) Biochem. Biophys. Res. Commun. 324, 269-275].
Assuntos
Antineoplásicos , Neoplasias da Mama/metabolismo , Citotoxinas , Piruvatos , Microambiente Tumoral/efeitos dos fármacos , Antineoplásicos/farmacocinética , Antineoplásicos/farmacologia , Basigina/metabolismo , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Citotoxinas/farmacocinética , Citotoxinas/farmacologia , Inibidores Enzimáticos/farmacocinética , Inibidores Enzimáticos/farmacologia , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Glicosilação/efeitos dos fármacos , Humanos , Concentração de Íons de Hidrogênio , Transportadores de Ácidos Monocarboxílicos/metabolismo , Proteínas Musculares/metabolismo , Proteínas de Neoplasias/metabolismo , Transporte Proteico/efeitos dos fármacos , Piruvatos/farmacocinética , Piruvatos/farmacologia , Simportadores/metabolismoRESUMO
Clinical research is currently exploring the validity of the anti-tumor candidate 3-bromopyruvate (3-BP) as a novel treatment for several types of cancer. However, recent publications have overlooked rarely-cited earlier work about the instability of 3-BP and its decay to 3-hydroxypyruvate (3-HP) which have obvious implications for its mechanism of action against tumors, how it is administered, and for precautions when preparing solutions of 3-BP. This study found the first-order decay rate of 3-BP at physiological temperature and pH has a half-life of only 77 min. Lower buffer pH decreases the decay rate, while choice of buffer and concentration do not affect it. A method for preparing more stable solutions is also reported.
Assuntos
Antineoplásicos/uso terapêutico , Piruvatos/uso terapêutico , Antineoplásicos/farmacocinética , Meia-Vida , Concentração de Íons de Hidrogênio , Espectroscopia de Ressonância Magnética , Piruvatos/farmacocinéticaRESUMO
We have investigated the cytotoxicity in Saccharomyces cerevisiae of the novel antitumor agent 3-bromopyruvate (3-BP). 3-BP enters the yeast cells through the lactate/pyruvate H(+) symporter Jen1p and inhibits cell growth at minimal inhibitory concentration of 1.8 mM when grown on non-glucose conditions. It is not submitted to the efflux pumps conferring Pleiotropic Drug Resistance in yeast. Yeast growth is more sensitive to 3-BP than Gleevec (Imatinib methanesulfonate) which in contrast to 3-BP is submitted to the PDR network of efflux pumps. The sensitivity of yeast to 3-BP is increased considerably by mutations or chemical treatment by buthionine sulfoximine that decrease the intracellular concentration of glutathione.
Assuntos
Antineoplásicos Alquilantes/farmacocinética , Antineoplásicos Alquilantes/toxicidade , Piruvatos/farmacocinética , Piruvatos/toxicidade , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/metabolismo , Antineoplásicos Alquilantes/metabolismo , Butionina Sulfoximina/farmacologia , Glutationa/metabolismo , Testes de Sensibilidade Microbiana , Transportadores de Ácidos Monocarboxílicos/metabolismo , Piruvatos/metabolismo , Saccharomyces cerevisiae/crescimento & desenvolvimento , Proteínas de Saccharomyces cerevisiae/metabolismo , Simportadores/metabolismoRESUMO
The aim of the present study was to evaluate in a mouse model of spinal cord injury (SCI) the effect of the treatment with ethyl pyruvate (EP). Spinal cord injury was induced by the application of vascular clips (force of 24 g) to the dura via a four-level T5-T8 laminectomy in mice. Treatment with EP (75, 25, or 8.5 mg/kg) 1 and 6 h after the SCI significantly decreased (a) the degree of spinal cord inflammation and tissue injury (histological score), (b) neutrophil infiltration (myeloperoxidase activity), (c) nitrotyrosine formation and iNOS expression, (d) proinflammatory cytokines expression, (e) nuclear factor kappaB activation, (f) extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase phosphorylation, and (g) apoptosis (TUNEL staining, Fas ligand, Bax, and Bcl-2 expression). Moreover, EP (75, 25, or 8.5 mg/kg) significantly ameliorated in a dose-dependent manner the loss of limb function (evaluated by motor recovery score). Taken together, our results demonstrate that EP treatment reduces the development of inflammation and tissue injury associated with spinal cord trauma.
Assuntos
Piruvatos/farmacocinética , Traumatismos da Medula Espinal/tratamento farmacológico , Animais , Apoptose/efeitos dos fármacos , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Proteína Ligante Fas/biossíntese , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Inflamação/patologia , Masculino , Camundongos , Proteína Quinase 1 Ativada por Mitógeno/biossíntese , Proteína Quinase 3 Ativada por Mitógeno/biossíntese , Infiltração de Neutrófilos/efeitos dos fármacos , Óxido Nítrico Sintase Tipo II/metabolismo , Peroxidase/metabolismo , Fosforilação , Traumatismos da Medula Espinal/metabolismo , Traumatismos da Medula Espinal/patologia , Fatores de Tempo , Tirosina/análogos & derivados , Tirosina/metabolismo , Proteína X Associada a bcl-2/biossínteseRESUMO
The aim of this study was to determine the biodistribution and tumor targeting ability of (14)C-labeled 3-bromopyruvate ([(14)C]3-BrPA) after i.a. and i.v. delivery in the VX2 rabbit model. In addition, we evaluated the effects of [(14)C]3-BrPA on tumor and healthy tissue glucose metabolism by determining (18)F-deoxyglucose (FDG) uptake. Last, we determined the survival benefit of i.a. administered 3-BrPA. In total, 60 rabbits with VX2 liver tumor received either 1.75 mM [(14)C]3-BrPA i.a., 1.75 mM [(14)C]3-BrPA i.v., 20 mM [(14)C]3-BrPA i.v., or 25 ml of phosphate-buffered saline (PBS). All rabbits (with the exception of the 20 mM i.v. group) received FDG 1 h before sacrifice. Next, we compared survival of animals treated with i.a. administered 1.75 mM [(14)C]3-BrPA in 25 ml of PBS (n = 22) with controls (n = 10). After i.a. infusion, tumor uptake of [(14)C]3-BrPA was 1.8 +/- 0.2% percentage of injected dose per gram of tissue (%ID/g), whereas other tissues showed minimal uptake. After i.v. infusion (1.75 mM), tumor uptake of [(14)C]3-BrPA was 0.03 +/- 0.01% ID/g. After i.a. administration of [(14)C]3-BrPA, tumor uptake of FDG was 26 times lower than in controls. After i.v. administration of [(14)C]3-BrPA, there was no significant difference in tumor FDG uptake. Survival analysis showed that rabbits treated with 1.75 mM 3-BrPA survived longer (55 days) than controls (18.6 days). Intra-arterially delivered 3-BrPA has a favorable biodistribution profile, combining a high tumor uptake resulting in blockage of FDG uptake with no effects on healthy tissue. The local control of the liver tumor by 3-BrPA resulted in a significant survival benefit.
Assuntos
Infusões Intra-Arteriais , Neoplasias Hepáticas Experimentais/tratamento farmacológico , Piruvatos/administração & dosagem , Piruvatos/farmacocinética , Animais , Radioisótopos de Carbono , Fluordesoxiglucose F18 , Neoplasias Hepáticas Experimentais/mortalidade , Coelhos , Distribuição TecidualRESUMO
Hypoxia stimulates hepatocellular carcinoma (HCC) cell growth via hexokinase (HK) II induction, and alternatively, HK II inhibition induces apoptosis by activating mitochondrial signaling. This study was to investigate whether the induction of HK II by hypoxia is associated with enhanced mitochondrial stability and to confirm the apoptosis-inducing efficacy of HK II inhibitor in an in vivo model of HCC. Mitochondrial stability was examined by treating isolated mitochondria with deoxycholate, a permeability-enhancing agent. Alteration of permeability transition pore complex composition was analyzed by immunoprecipitation and immunoblotting. An in vivo model of HCC was established in C3H mice i.d. implanted with MH134 cells. The antitumor efficacy of i.p. given 3-bromopyruvate (3-BrPA), a HK II inhibitor, was evaluated by measuring tumor volumes and quantifying apoptosis using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining and (99m)Tc-hydrazinonicotinamide-Annexin V scans. Hypoxia enhanced mitochondrial stability, and this was inhibited by 3-BrPA treatment. In particular, HK II levels in permeability transition pore complex immunoprecipitates were reduced after 3-BrPA treatment. In mice treated with 3-BrPA, mean tumor volumes and tumor volume growth were found to be significantly reduced. Moreover, percentages of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling-positive cells were significantly increased in 3-BrPA-treated mice, and this apoptosis-inducing efficacy was reflected in vivo by (99m)Tc-hydrazinonicotinamide-Annexin V imaging. Our results show that hypoxia enhances mitochondrial stability via HK II induction and that HK II inhibitor treatment exhibits an in vivo antitumor effect by inducing apoptosis. Therefore, HK II inhibitors may be therapeutically useful for the treatment of advanced infiltrative hypovascular HCCs, which are growing in a hypoxic environment.
Assuntos
Apoptose/efeitos dos fármacos , Carcinoma Hepatocelular/tratamento farmacológico , Inibidores Enzimáticos/farmacologia , Hexoquinase/antagonistas & inibidores , Neoplasias Hepáticas Experimentais/tratamento farmacológico , Piruvatos/farmacologia , Animais , Anexina A5/metabolismo , Azirinas , Carcinoma Hepatocelular/enzimologia , Carcinoma Hepatocelular/metabolismo , Inibidores Enzimáticos/farmacocinética , Citometria de Fluxo , Hexoquinase/metabolismo , Humanos , Hipóxia , Immunoblotting , Imunoprecipitação , Neoplasias Hepáticas Experimentais/enzimologia , Neoplasias Hepáticas Experimentais/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C3H , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Fosfatidilcolinas , Complexo Piruvato Desidrogenase/antagonistas & inibidores , Piruvatos/farmacocinética , Distribuição Tecidual , Células Tumorais CultivadasRESUMO
Studies describe an attenuation of sugar cataract formation by topical administration of ethyl pyruvate. Cataract formation was induced by feeding young rats a 30% galactose diet. Mature cataracts appeared in about thirty days. Instillation of the eye drops containing 5% ethyl pyruvate decelerated the process significantly. Biochemically, the effect was reflected by lowering in the contents of dulcitol and glycated proteins. The ATP levels were also higher in comparison to the placebo treated group. The effects are hence attributable to the effect of pyruvate in inhibiting dulcitol synthesis and protein glycation, in addition to its antioxidant properties and metabolic support. The use of esterified pyruvate instead of the unesterified pyruvate was preferred because of its greater penetration through the cornea and consequently a higher concentration attained in the aqueous humor.
Assuntos
Catarata/prevenção & controle , Piruvatos/farmacologia , Trifosfato de Adenosina/metabolismo , Animais , Humor Aquoso/metabolismo , Catarata/etiologia , Catarata/metabolismo , Cristalinas/metabolismo , Sacarose Alimentar/administração & dosagem , Galactitol/metabolismo , Galactose/administração & dosagem , Galactosemias/complicações , Glicosilação , Masculino , Soluções Oftálmicas , Piruvatos/administração & dosagem , Piruvatos/farmacocinética , Ratos , Ratos Sprague-DawleyRESUMO
We examined pyruvate turnover using 1-[11C]pyruvate in the brain and epicranial muscle of 6 patients with mitochondrial encephalomyopathy (MEM), diagnosed by muscle biopsy and mitochondrial enzyme assay. The radioactivity was measured by positron emission tomography (PET). The time-activity curve for 11C in both brain and muscle generated after i.v. injection of 1-[11C]pyruvate consisted of 2 components in normal subjects and patients, i.e. a fast and a slow component which were assumed to represent the aerobic (mitochondrial) and anaerobic (glycolytic) metabolism of pyruvate, respectively. In the brain and muscle of patients, the aerobic component was smaller and the anaerobic larger than in normals. The extent of this abnormality seemed to reflect the severity of the disease. The same slight abnormality for [11C]pyruvate turnover was also observed in the brain of MEM patients who were without cerebral symptoms. Cerebral blood flow (CBF) and cerebral oxygen consumption (CMRO2) of most patients were lower than those of normals, and the oxygen extraction fraction (OEF) was decreased in many patients.
Assuntos
Encefalopatias Metabólicas/metabolismo , Encéfalo/metabolismo , Mitocôndrias/patologia , Músculos/metabolismo , Doenças Musculares/metabolismo , Piruvatos/farmacocinética , Tomografia Computadorizada de Emissão , Adulto , Encéfalo/diagnóstico por imagem , Encefalopatias Metabólicas/diagnóstico por imagem , Encefalopatias Metabólicas/genética , Circulação Cerebrovascular , Metabolismo Energético , Humanos , Mitocôndrias/metabolismo , Músculos/diagnóstico por imagem , Doenças Musculares/diagnóstico por imagem , Doenças Musculares/genética , Consumo de OxigênioRESUMO
Two lines of mouse Ehrlich ascites cancer differing in the rate of aerobic glycolysis were employed. Both lines use lactate and show oxidizing action upon cytosol NADH. The obtained results indicate the possibility that cytosol NADH reoxidation is partly independent of the mitochondrial respiratory system.