RESUMEN
Anticancer effects of a common lipid-lowering drug, fenofibrate, have been described in the literature for a quite some time; however, fenofibrate has not been used as a direct anticancer therapy. We have previously reported that fenofibrate in its unprocessed form (ester) accumulates in the mitochondria, inhibits mitochondrial respiration, and triggers a severe energy deficit and extensive glioblastoma cell death. However, fenofibrate does not cross the blood brain barrier and is quickly processed by blood and tissue esterases to form the PPARα agonist fenofibric acid, which is practically ineffective effective in triggering cancer cell death. To address these issues, we have made several chemical modifications in fenofibrate structure to increase its stability, water solubility, tissue penetration, and ultimately anticancer potential. Our data show that, in comparison to fenofibrate, four new compounds designated here as PP1, PP2, PP3, and PP4 have improved anticancer activity in vitro. Like fenofibrate, the compounds block mitochondrial respiration and trigger massive glioblastoma cell death in vitro. In addition, one of the lead compounds, PP1, has improved water solubility and is significantly more stable when exposed to human blood in comparison to fenofibrate. Importantly, mice bearing large intracranial glioblastoma tumors demonstrated extensive areas of tumor cell death within the tumor mass following oral administration of PP1, and the treated mice did not show any major signs of distress, and accumulated PP1 at therapeutically relevant concentrations in several tissues, including brain and intracranial tumors.
RESUMEN
BACKGROUND: Increasing evidence has shown the close link between energy metabolism and the differentiation, function, and longevity of immune cells. Chronic inflammatory conditions such as parasitic infections and cancer trigger a metabolic reprogramming from the preferential use of glucose to the up-regulation of fatty acid oxidation (FAO) in myeloid cells, including macrophages and granulocytic and monocytic myeloid-derived suppressor cells. Asthma is a chronic inflammatory condition where macrophages, eosinophils, and polymorphonuclear cells play an important role in its pathophysiology. OBJECTIVE: We tested whether FAO might play a role in the development of asthma-like traits and whether the inhibition of this metabolic pathway could represent a novel therapeutic approach. METHODS: OVA- and house dust mite (HDM)-induced murine asthma models were used in this study. RESULTS: Key FAO enzymes were significantly increased in the bronchial epithelium and inflammatory immune cells infiltrating the respiratory epithelium of mice exposed to OVA or HDM. Pharmacologic inhibition of FAO significantly decreased allergen-induced airway hyperresponsiveness, decreased the number of inflammatory cells, and reduced the production of cytokines and chemokines associated with asthma. CONCLUSIONS AND CLINICAL RELEVANCE: These novel observations suggest that allergic airway inflammation increases FAO in inflammatory cells to support the production of cytokines, chemokines, and other factors important in the development of asthma. Inhibition of FAO by re-purposing existing drugs approved for the treatment of heart disease may provide a novel therapeutic approach for the treatment of asthma.
Asunto(s)
Asma/etiología , Asma/metabolismo , Ácidos Grasos/metabolismo , Sistema Inmunológico/citología , Sistema Inmunológico/inmunología , Sistema Inmunológico/metabolismo , Oxidación-Reducción , Alérgenos , Animales , Asma/tratamiento farmacológico , Asma/patología , Biomarcadores , Quimiocinas/metabolismo , Citocinas/metabolismo , Modelos Animales de Enfermedad , Sistema Inmunológico/efectos de los fármacos , Inmunidad Innata/inmunología , Inmunoglobulina E/inmunología , Masculino , Ratones , Terapia Molecular Dirigida , Hipersensibilidad Respiratoria/inmunología , Hipersensibilidad Respiratoria/metabolismo , Hipersensibilidad Respiratoria/patologíaRESUMEN
Fenofibrate, a well-known normolipidemic drug, has been shown to exert strong anticancer effects against tumors of neuroectodermal origin including glioblastoma. Although some pharmacokinetic studies were performed in the past, data are still needed about the detailed subcellular and tissue distribution of fenofibrate (FF) and its active metabolite, fenofibric acid (FA), especially in respect to the treatment of intracranial tumors. We used high performance liquid chromatography (HPLC) to elucidate the intracellular, tissue and body fluid distribution of FF and FA after oral administration of the drug to mice bearing intracranial glioblastoma. Following the treatment, FF was quickly cleaved to FA by blood esterases and FA was detected in the blood, urine, liver, kidney, spleen and lungs. We have also detected small amounts of FA in the brains of two out of six mice, but not in the brain tumor tissue. The lack of FF and FA in the intracranial tumors prompted us to develop a new method for intracranial delivery of FF. We have prepared and tested in vitro biodegradable poly-lactic-co-glycolic acid (PLGA) polymer wafers containing FF, which could ultimately be inserted into the brain cavity following resection of the brain tumor. HPLC-based analysis demonstrated a slow and constant diffusion of FF from the wafer, and the released FF abolished clonogenic growth of glioblastoma cells. On the intracellular level, FF and FA were both present in the cytosolic fraction. Surprisingly, we also detected FF, but not FA in the cell membrane fraction. Electron paramagnetic resonance spectroscopy applied to spin-labeled phospholipid model-membranes revealed broadening of lipid phase transitions and decrease of membrane polarity induced by fenofibrate. Our results indicate that the membrane-bound FF could contribute to its exceptional anticancer potential in comparison to other lipid-lowering drugs, and advocate for intracranial delivery of FF in the combined pharmacotherapy against glioblastoma.
Asunto(s)
Plásticos Biodegradables/farmacocinética , Neoplasias Encefálicas/tratamiento farmacológico , Encéfalo/metabolismo , Portadores de Fármacos/farmacocinética , Fenofibrato/análogos & derivados , Glioblastoma/tratamiento farmacológico , Animales , Encéfalo/efectos de los fármacos , Línea Celular Tumoral , Femenino , Fenofibrato/farmacocinética , Fenofibrato/farmacología , Humanos , Ácido Láctico/farmacocinética , Ratones , Ratones Desnudos , Ácido Poliglicólico/farmacocinética , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Polímeros/farmacocinética , Distribución TisularRESUMEN
New treatments for adults with acute lymphoblastic T-cell leukemia (T-ALL) are urgently needed, as the current rate of overall remission in these patients is only about 40 percent. We recently showed the potential therapeutic benefit of the pegylated-human-arginase I (peg-Arg I) in T-ALL. However, the mechanisms by which peg-Arg I induces an anti-T-ALL effect remained unknown. Our results show the induction of T-ALL cell apoptosis by peg-Arg I, which associated with a global arrest in protein synthesis and with the phosphorylation of the eukaryotic-translation-initiation factor 2 alpha (eIF2α). Inhibition of eIF2α phosphorylation in T-ALL cells prevented the apoptosis induced by peg-Arg I, whereas the expression of a phosphomimetic eIF2α form increased the sensibility of T-ALL cells to peg-Arg I. Phosphorylation of eIF2α by peg-Arg I was mediated through kinases PERK and GCN2 and down-regulation of phosphatase GADD34. GCN2 and decreased GADD34 promoted T-ALL cell apoptosis after treatment with peg-Arg I, whereas PERK had an unexpected anti-apoptotic role. Additional results showed that phospho-eIF2α signaling further increased the anti-leukemic effects induced by peg-Arg I in T-ALL-bearing mice. These results suggest the central role of phospho-eIF2α in the anti-T-ALL effects induced by peg-Arg I and support its study as a therapeutic target.
Asunto(s)
Arginasa/administración & dosificación , Factor 2 Eucariótico de Iniciación/metabolismo , Polietilenglicoles/química , Leucemia-Linfoma Linfoblástico de Células T Precursoras/prevención & control , Proteína Fosfatasa 1/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , eIF-2 Quinasa/metabolismo , Animales , Apoptosis/efectos de los fármacos , Western Blotting , Proliferación Celular/efectos de los fármacos , Femenino , Técnica del Anticuerpo Fluorescente , Humanos , Ratones , Ratones Endogámicos NOD , Ratones SCID , Fosforilación , Leucemia-Linfoma Linfoblástico de Células T Precursoras/metabolismo , Leucemia-Linfoma Linfoblástico de Células T Precursoras/patología , Proteínas Recombinantes/uso terapéutico , Transducción de Señal , Tasa de SupervivenciaAsunto(s)
Antimetabolitos Antineoplásicos/farmacología , Azacitidina/análogos & derivados , Azacitidina/farmacología , Metilación de ADN/efectos de los fármacos , Metilasas de Modificación del ADN/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , Ácido Butírico/farmacología , Diferenciación Celular/efectos de los fármacos , Islas de CpG , Citosina/metabolismo , Decitabina , Humanos , Células K562Asunto(s)
Antineoplásicos/farmacología , Azacitidina/análogos & derivados , ADN (Citosina-5-)-Metiltransferasas/antagonistas & inhibidores , Metilación de ADN/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Vidarabina/farmacología , Antimetabolitos Antineoplásicos/farmacología , Azacitidina/farmacología , Línea Celular , Supervivencia Celular/efectos de los fármacos , Decitabina , Relación Dosis-Respuesta a Droga , Humanos , Vidarabina/análogos & derivadosRESUMEN
Variants of the human K562 were developed against the nucleoside analogues cytosine arabinoside, 2 chlorodeoxyadenosine, fludarabine and gemcitabine. The resistant lines displayed a high degree of cross-resistance to all nucleoside analogues, with little or no cross resistance to other agents. There was a profound accumulation defect of the different nucleoside analogues in all of the variants. There was a strong overexpression of 5'nucleotidase, measured by rt-PCR and enzyme activity, in all resistant variants. There was a two fold increase of ribonucleotide reductase in the fludarabine resistant line and increased expression of purine nucleoside phosphorylase in the 2 chlorodeoxyadenosine selected line. Karyotypic analysis revealed the loss of a 6(q16;q22) deletion present in the parental line in all of the resistant lines. This portion of chromosome 6 has been shown to contain the gene for 5'nucleotidase. Early events in the transport and metabolism appear to be involved in the resistance mechanisms to nucleoside analogues and are responsible for broad cross resistance to this family of compounds.
Asunto(s)
Antimetabolitos Antineoplásicos/toxicidad , Resistencia a Múltiples Medicamentos , Células K562/efectos de los fármacos , 5'-Nucleotidasa/metabolismo , Adenosina Desaminasa/metabolismo , División Celular/efectos de los fármacos , Células Clonales , Resistencia a Antineoplásicos , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Purina-Nucleósido Fosforilasa/metabolismo , Ribonucleótido Reductasas/metabolismoRESUMEN
Resistant variants of the human leukaemic line K562 were developed using selection with the deoxynucleoside analogues cytosine arabinoside, 2-chlorodeoxyadenosine, fludarabine and gemcitabine. The resistant lines displayed a high degree of cross resistance to all deoxynucleoside analogues, with little or no cross resistance to other agents. There was a profound accumulation defect of all nucleoside analogues in the resistant variants but no significant defect in nucleoside transport in any of the variants. 5' nucleotidase activity was strongly increased and deoxycytidine kinase activity was moderately reduced in all of the resistant variants, resulting in reduced accumulation of triphosphate analogues. In addition a deletion in one of the alleles of the deoxycytidine kinase was detected in the fludarabine-resistant line. Ribonucleotide reductase activity was found to be strongly increased in the gemcitabine-selected line and purine nucleoside phosphorylase was increased in the 2-chlorodeoxyadenosine-selected line. Free nucleotide pools were increased in the 2-chlorodeoxyadenosine-selected line. There was no expression of the mdr1 gene by the resistant lines. Karyotypic analysis and FISH experiments using a 6q21 specific probe showed alterations in the 6(q16-q22) region which contains the 5'-nucleotidase gene. Early events in the activation and degradation of deoxynucleoside analogues appear to constitute common mechanisms of resistance to these compounds.
Asunto(s)
Antineoplásicos/uso terapéutico , Cladribina/uso terapéutico , Citarabina/uso terapéutico , Desoxicitidina/análogos & derivados , Leucemia Eritroblástica Aguda/tratamiento farmacológico , Vidarabina/análogos & derivados , Desoxicitidina/uso terapéutico , Desoxicitidina Quinasa/metabolismo , Resistencia a Antineoplásicos , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Células K562 , Leucemia Eritroblástica Aguda/enzimología , Fenotipo , Vidarabina/uso terapéutico , GemcitabinaRESUMEN
The effect of 2-chloro-2'-deoxyadenosine and 9-beta-D-arabinosyl-2-fluoroadenine on DNA methyltransferase activity in stimulated human T-lymphocytes was estimated. In comparative studies 5-aza-deoxycytidine and deoxyadenosine plus deoxycoformycin were used. These antileukemic compounds demonstrated different effects; both 2CdA and dAdo plus dCF, like 5-aza-dCyt, inhibited the enzyme activity by 85-90% after 72 hours activation of lymphocytes, while the effect of F-ara-A, under the same conditions, was insignificant.
Asunto(s)
Cladribina/farmacología , ADN (Citosina-5-)-Metiltransferasas/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , Linfocitos T/efectos de los fármacos , Vidarabina/análogos & derivados , Humanos , Linfocitos T/enzimología , Vidarabina/farmacologíaAsunto(s)
Cladribina/farmacología , ADN (Citosina-5-)-Metiltransferasas/metabolismo , Hidrolasas/metabolismo , Leucemia L1210/enzimología , Adenosilhomocisteinasa , Animales , Antimetabolitos Antineoplásicos/farmacología , Antineoplásicos/farmacología , Azacitidina/análogos & derivados , Azacitidina/farmacología , Decitabina , Cinética , Ratones , Células Tumorales Cultivadas , Vidarabina/análogos & derivados , Vidarabina/farmacologíaRESUMEN
The studies on the metabolism and toxic mechanism of 2-chloro-2'-deoxyadenosine (2CdA, Cladribine), a new antileukemic drug, were reviewed. 2CdA, being a 2-halogenated, adenosine deaminase-resistant analogue of deoxyadenosine, is phosphorylated to the mono-, di, and triphosphate chlorodeoxy adenosine and the first step of phosphorylation is taken in the presence of enzymes, mainly kinase deoxycytidine (although in mitochondria it is phosphorylated by kinase deoxyguanosine). Triphosphate derivative of 2CdA is commonly considered to be the agent inducing cell apoptosis resulting from inhibition of ribonucleotide reductase, DNA polymerases and DNA repair. Recent studies on toxicity of 2CdA showed that the nucleoside possesses inhibitory activity against enzymes which are responsible for metabolism of deoxyadenosine, which suggests that the mechanism of toxicity by 2CdA includes a block in dAdo metabolic pathways which is very important for normal function of immune system cells. The agent under discussion and two other adenosine analogues (i.e. fludarabine and 2'-deoxycoformycin) which exhibit cytotoxicity against dividing and resting lymphocytes revolutionized the treatment of indolent lymphoid malignancies (i.e. chronic lymphocytic leukemia, non-Hodgkin's lymphoma, cutaneous T cell lymphoma and hairy cell leukemia). Particularly, in the treatment of hairy cell leukemia, 2-chloro-2'-deoxyadenosine demonstrated excellent efficacy, achieved after a single 7-day course, with an acceptable tolerability profile, suggesting that cladribine is likely to be more effective than other agents recommended in this disease. Preliminary clinical data, extremely encouraging in the case of 2CdA indicate that biomolecular mechanisms of the drug cytotoxicity is worth wide presentation.