RESUMEN
BACKGROUND: Acylcarnitine is an intermediate product of fatty acid oxidation. It is reported to be closely associated with the occurrence of diabetic cardiomyopathy (DCM). However, the mechanism of acylcarnitine affecting myocardial disorders is yet to be explored. This current research explores the different chain lengths of acylcarnitines as biomarkers for the early diagnosis of DCM and the mechanism of acylcarnitines for the development of DCM in-vitro. METHODS: In a retrospective non-interventional study, 50 simple type 2 diabetes mellitus patients and 50 DCM patients were recruited. Plasma samples from both groups were analyzed by high throughput metabolomics and cluster heat map using mass spectrometry. Principal component analysis was used to compare the changes occurring in the studied 25 acylcarnitines. Multivariable binary logistic regression was used to analyze the odds ratio of each group for factors and the 95% confidence interval in DCM. Myristoylcarnitine (C14) exogenous intervention was given to H9c2 cells to verify the expression of lipid metabolism-related protein, inflammation-related protein expression, apoptosis-related protein expression, and cardiomyocyte hypertrophy and fibrosis-related protein expression. RESULTS: Factor 1 (C14, lauroylcarnitine, tetradecanoyldiacylcarnitine, 3-hydroxyl-tetradecanoylcarnitine, arachidic carnitine, octadecanoylcarnitine, 3-hydroxypalmitoleylcarnitine) and factor 4 (octanoylcarnitine, hexanoylcarnitine, decanoylcarnitine) were positively correlated with the risk of DCM. Exogenous C14 supplementation to cardiomyocytes led to increased lipid deposition in cardiomyocytes along with the obstacles in adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) signaling pathways and affecting fatty acid oxidation. This further caused myocardial lipotoxicity, ultimately leading to cardiomyocyte hypertrophy, fibrotic remodeling, and increased apoptosis. However, this effect was mitigated by the AMPK agonist acadesine. CONCLUSIONS: The increased plasma levels in medium and long-chain acylcarnitine extracted from factors 1 and 4 are closely related to the risk of DCM, indicating that these factors can be an important tool for DCM risk assessment. C14 supplementation associated lipid accumulation by inhibiting the AMPK/ACC/CPT1 signaling pathway, aggravated myocardial lipotoxicity, increased apoptosis apart from cardiomyocyte hypertrophy and fibrosis were alleviated by the acadesine.
Asunto(s)
Carnitina/análogos & derivados , Diabetes Mellitus Tipo 2/complicaciones , Cardiomiopatías Diabéticas/metabolismo , Metabolismo de los Lípidos , Adulto , Aminoimidazol Carboxamida/análogos & derivados , Aminoimidazol Carboxamida/farmacología , Animales , Biomarcadores/sangre , Carnitina/sangre , Carnitina/química , Carnitina/farmacología , Línea Celular , Diabetes Mellitus Tipo 2/metabolismo , Femenino , Humanos , Metabolismo de los Lípidos/efectos de los fármacos , Masculino , Espectrometría de Masas , Persona de Mediana Edad , Mioblastos Cardíacos/efectos de los fármacos , Mioblastos Cardíacos/metabolismo , Ácidos Mirísticos/farmacología , Ratas , Estudios Retrospectivos , Ribonucleósidos/farmacología , Factores de RiesgoRESUMEN
BACKGROUND: On March 11, 2020, the World Health Organization (WHO) declared the outbreak of coronavirus disease (COVID-19) a pandemic. Since then, thousands of people have suffered and died, making the need for a treatment of severe acute respiratory syndrome-related coronavirus-2 (SARS-CoV-2) more crucial than ever. MATERIALS AND METHODS: The authors carried out a search in PubMed, ClinicalTrials.gov and New England Journal of Medicine (NEJM) for COVID-19 to provide information on the most promising treatments against SARS-CoV-2. RESULTS: Possible COVID-19 agents with promising efficacy and favorable safety profile were identified. The results support the combination of copper, N-acetylcysteine (NAC), colchicine and nitric oxide (NO) with candidate antiviral agents, remdesivir or EIDD-2801, as a treatment for patients positive for SARS-CoV-2. CONCLUSION: The authors propose to study the effects of the combination of copper, NAC, colchicine, NO and currently used experimental antiviral agents, remdesivir or EIDD-2801, as a potential treatment scheme for SARS-COV-2.
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Acetilcisteína/uso terapéutico , Adenosina Monofosfato/análogos & derivados , Alanina/análogos & derivados , Antivirales/uso terapéutico , Colchicina/uso terapéutico , Cobre/uso terapéutico , Infecciones por Coronavirus/tratamiento farmacológico , Óxido Nítrico/uso terapéutico , Neumonía Viral/tratamiento farmacológico , Ribonucleósidos/uso terapéutico , Acetilcisteína/administración & dosificación , Acetilcisteína/farmacología , Adenosina Monofosfato/administración & dosificación , Adenosina Monofosfato/farmacología , Adenosina Monofosfato/uso terapéutico , Adyuvantes Inmunológicos/administración & dosificación , Adyuvantes Inmunológicos/uso terapéutico , Alanina/administración & dosificación , Alanina/farmacología , Alanina/uso terapéutico , Antiinflamatorios/administración & dosificación , Antiinflamatorios/uso terapéutico , Antivirales/administración & dosificación , Antivirales/farmacología , Autofagia/efectos de los fármacos , Betacoronavirus/efectos de los fármacos , Betacoronavirus/fisiología , COVID-19 , Colchicina/administración & dosificación , Colchicina/farmacología , Cobre/administración & dosificación , Infecciones por Coronavirus/inmunología , Infecciones por Coronavirus/fisiopatología , Citidina/análogos & derivados , Sinergismo Farmacológico , Quimioterapia Combinada , Humanos , Hidroxilaminas , Inflamación , Óxido Nítrico/administración & dosificación , Óxido Nítrico/farmacología , Pandemias , Neumonía Viral/inmunología , Neumonía Viral/fisiopatología , Profármacos/administración & dosificación , Profármacos/uso terapéutico , Ribonucleósidos/administración & dosificación , Ribonucleósidos/farmacología , SARS-CoV-2 , Internalización del Virus/efectos de los fármacos , Replicación Viral/efectos de los fármacosRESUMEN
An effective treatment for non-alcoholic fatty liver disease (NAFLD) is urgently needed. In the present study, we investigated whether the Chinese medicine Chai Hu Li Zhong Tang (CHLZT) could protect against the development of NAFLD. Rats in an animal model of NAFLD were treated with CHLZT, and their serum levels of cholesterol (TC), triglycerides (TG), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) were detected with an automatic biochemical analyzer. A cellular model of NAFLD was also established by culturing HepG2 cells in a medium that contained a long chain fat emulsion. Those cells were treated with CHLZT that contained serum from rats. After treatment, the levels of adenylate-activated protein kinase (AMPK) α (AMPKα), p-AMPKα, acetyl coenzyme A carboxylase (ACC) α (ACCα), pACCα, PPARγ, and SREBP-2 were detected. The AMPK agonist, acadesine (AICAR), was used as a positive control compound. Our results showed that CHLZT or AICAR significantly decreased the serum levels of TG, TC, LDL-C, AST, ALT, and insulin in NAFLD rats, and significantly increased their serum HDL-C levels. Treatments with CHLZT or AICAR significantly decreased the numbers of lipid droplets in NAFLD liver tissues and HepG2 cells. CHLZT and AICAR increased the levels of p-AMPKα and PPARγ in the NAFLD liver tissues and HepG2 cells, but decreased the levels of ACC-α, p-ACC-α, SREBP-2, and 3-hydroxyl-3-methylglutaryl-coenzyme A reductase (HMGR). CHLZT protects against NAFLD by activating AMPKα, and also by inhibiting ACC activity, down-regulating SREBP2 and HMGR, and up-regulating PPAR-γ. Our results suggest that CHLZT might be useful for treating NAFLD in the clinic.
Asunto(s)
Proteínas Quinasas Activadas por AMP/genética , Bupleurum/química , Medicamentos Herbarios Chinos/administración & dosificación , Enfermedad del Hígado Graso no Alcohólico/tratamiento farmacológico , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/genética , Proteínas Quinasas Activadas por AMP/sangre , Acetil-CoA Carboxilasa/sangre , Alanina Transaminasa/sangre , Aminoimidazol Carboxamida/análogos & derivados , Aminoimidazol Carboxamida/farmacología , Animales , Aspartato Aminotransferasas/sangre , HDL-Colesterol/sangre , LDL-Colesterol/sangre , Medicamentos Herbarios Chinos/química , Regulación de la Expresión Génica/efectos de los fármacos , Células Hep G2 , Humanos , Medicina Tradicional China/métodos , Enfermedad del Hígado Graso no Alcohólico/sangre , Enfermedad del Hígado Graso no Alcohólico/genética , Enfermedad del Hígado Graso no Alcohólico/patología , Ratas , Ribonucleósidos/farmacología , Proteína 2 de Unión a Elementos Reguladores de Esteroles/genética , Triglicéridos/sangreRESUMEN
BACKGROUND: Berberine, a well-known component of the Chinese herbal medicine Huanglian, has wide range of biochemical and pharmacological effects, including antineoplastic effect, but the exact mechanisms remain unclear. The aim of the present study was to evaluate the potential chemo-sensitization effect of berberine in ovarian cancer cell line A2780. METHODS: The expression of miR-93 was measure by RT-PCR. The target of miR-93 was confirmed by luciferase activity assay. Hoechst 33258 staining, Annexin V and PI double staining were used for apoptosis analysis. RESULTS: In this study, we found A2780/DDP cells that were incubated with berberine combined with cisplatin had a significantly lower survival than the control group. Berberine enhanced cisplatin induced apoptosis and induced G0/G1 cell cycle arrest in A2780 cells. Next, we observed that the miR-93 levels in cisplatin resistant cell lines were higher than that in cisplatin sensitive cell lines. Furthermore, our study found berberine could inhibit miR-93 expression and function in ovarian cancer, as shown by an increase of its target PTEN, an important tumor suppressor in ovarian cancer. A2780 cells that were treated with PTEN siRNA had increased survival compared to NC group and this could be partly alleviated by the AKT inhibitor Triciribine. More importantly, A2780 cells that were treated with PTEN siRNA had a survival pattern that is similar to cells with miR-93 overexpression. CONCLUSION: The results suggested that berberine modulated the sensitivity of cisplatin through miR-93/PTEN/AKT signaling pathway in the ovarian cancer cells.
Asunto(s)
Antineoplásicos/farmacología , Berberina/farmacología , Cisplatino/farmacología , Resistencia a Antineoplásicos/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica , MicroARNs/genética , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Resistencia a Antineoplásicos/genética , Sinergismo Farmacológico , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Células Epiteliales/patología , Femenino , Puntos de Control de la Fase G1 del Ciclo Celular/efectos de los fármacos , Genes Reporteros , Humanos , Luciferasas/genética , Luciferasas/metabolismo , MicroARNs/antagonistas & inhibidores , MicroARNs/metabolismo , Ovario/efectos de los fármacos , Ovario/metabolismo , Ovario/patología , Fosfohidrolasa PTEN/agonistas , Fosfohidrolasa PTEN/antagonistas & inhibidores , Fosfohidrolasa PTEN/genética , Fosfohidrolasa PTEN/metabolismo , Proteínas Proto-Oncogénicas c-akt/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-akt/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Ribonucleósidos/farmacología , Transducción de SeñalRESUMEN
Adenosine kinase (ADK) from Mycobacterium tuberculosis (Mtb) was selected as a target for design of antimycobacterial nucleosides. Screening of 7-(het)aryl-7-deazaadenine ribonucleosides with Mtb and human (h) ADKs and testing with wild-type and drug-resistant Mtb strains identified specific inhibitors of Mtb ADK with micromolar antimycobacterial activity and low cytotoxicity. X-ray structures of complexes of Mtb and hADKs with 7-ethynyl-7-deazaadenosine showed differences in inhibitor interactions in the adenosine binding sites. 1D (1)H STD NMR experiments revealed that these inhibitors are readily accommodated into the ATP and adenosine binding sites of Mtb ADK, whereas they bind preferentially into the adenosine site of hADK. Occupation of the Mtb ADK ATP site with inhibitors and formation of catalytically less competent semiopen conformation of MtbADK after inhibitor binding in the adenosine site explain the lack of phosphorylation of 7-substituted-7-deazaadenosines. Semiempirical quantum mechanical analysis confirmed different affinity of nucleosides for the Mtb ADK adenosine and ATP sites.
Asunto(s)
Adenosina Quinasa/antagonistas & inhibidores , Adenosina Quinasa/química , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/enzimología , Ribonucleósidos/química , Ribonucleósidos/farmacología , Adenina/análogos & derivados , Adenina/química , Adenosina Quinasa/metabolismo , Adenosina Trifosfato/metabolismo , Antituberculosos/química , Antituberculosos/farmacología , Sitios de Unión , Cristalografía por Rayos X , Evaluación Preclínica de Medicamentos , Humanos , Pruebas de Sensibilidad Microbiana , Resonancia Magnética Nuclear Biomolecular , Conformación Proteica , Relación Estructura-ActividadRESUMEN
The limited efficacy, in particular against the genotype 1 virus, as well as the variety of side effects associated with the current therapy for hepatitis C virus (HCV) infection necessitates more efficacious drugs. We found that phosphoramidate prodrugs of 2'-deoxy-2'-spirooxetane ribonucleosides form a novel class of HCV NS5B RNA-dependent RNA polymerase inhibitors, displaying EC50 values ranging from 0.2 to >98 µM, measured in the Huh7-replicon cell line, with no apparent cytotoxicity (CC50 > 98.4 µM). Confirming recent findings, the 2'-spirooxetane moiety was identified as a novel structural motif in the field of anti-HCV nucleosides. A convenient synthesis was developed that enabled the synthesis of a broad set of nucleotide prodrugs with varying substitution patterns. Extensive formation of the triphosphate metabolite was observed in both rat and human hepatocyte cultures. In addition, after oral dosing of several phosphoramidate derivatives of compound 21 to rats, substantial hepatic levels of the active triphosphate metabolite were found.
Asunto(s)
Antivirales/farmacología , Inhibidores Enzimáticos/farmacología , Hepacivirus/efectos de los fármacos , Profármacos/farmacología , Ribonucleósidos/farmacología , Proteínas no Estructurales Virales/antagonistas & inhibidores , Animales , Antivirales/química , Antivirales/farmacocinética , Área Bajo la Curva , Evaluación Preclínica de Medicamentos , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacocinética , Hepacivirus/enzimología , Humanos , Espectroscopía de Resonancia Magnética , Ratas , Ratas Sprague-Dawley , Ribonucleósidos/química , Ribonucleósidos/farmacocinéticaRESUMEN
African trypanosomes are capable of both de novo synthesis and salvage of pyrimidines. The last two steps in de novo synthesis are catalysed by UMP synthase (UMPS) - a bifunctional enzyme comprising orotate phosphoribosyl transferase (OPRT) and orotidine monophosphate decarboxylase (OMPDC). To investigate the essentiality of pyrimidine biosynthesis in Trypanosoma brucei, we generated a umps double knockout (DKO) line by gene replacement. The DKO was unable to grow in pyrimidine-depleted medium in vitro, unless supplemented with uracil, uridine, deoxyuridine or UMP. DKO parasites were completely resistant to 5-fluoroorotate and hypersensitive to 5-fluorouracil, consistent with loss of UMPS, but remained sensitive to pyrazofurin indicating that, unlike mammalian cells, the primary target of pyrazofurin is not OMPDC. The null mutant was unable to infect mice indicating that salvage of host pyrimidines is insufficient to support growth. However, following prolonged culture in vitro, parasites regained virulence in mice despite retaining pyrimidine auxotrophy. Unlike the wild-type, both pyrimidine auxotrophs secreted substantial quantities of orotate, significantly higher in the virulent DKO line. We propose that this may be responsible for the recovery of virulence in mice, due to host metabolism converting orotate to uridine, thereby bypassing the loss of UMPS in the parasite.
Asunto(s)
Complejos Multienzimáticos/genética , Orotato Fosforribosiltransferasa/genética , Orotidina-5'-Fosfato Descarboxilasa/genética , Pirimidinas/metabolismo , Trypanosoma brucei brucei/metabolismo , Trypanosoma brucei brucei/patogenicidad , Amidas , Animales , Transporte Biológico , Línea Celular , Desoxiuridina/metabolismo , Fluorouracilo/farmacología , Técnicas de Inactivación de Genes , Ratones/parasitología , Complejos Multienzimáticos/metabolismo , Orotato Fosforribosiltransferasa/metabolismo , Ácido Orótico/análogos & derivados , Ácido Orótico/metabolismo , Ácido Orótico/farmacología , Orotidina-5'-Fosfato Descarboxilasa/metabolismo , Pirazoles , Pirimidinas/biosíntesis , Ribonucleósidos/farmacología , Ribosa , Transfección , Tripanocidas/farmacología , Trypanosoma brucei brucei/efectos de los fármacos , Uracilo/metabolismo , Uridina/metabolismo , Uridina Monofosfato/metabolismo , VirulenciaRESUMEN
This study tested whether chronic systemic administration of 5-aminoimidazole-4-carboxamide-1-ß-D-ribofuranoside (AICAR) could attenuate hyperphagia, reduce lean and fat mass losses, and improve whole-body energy homeostasis in insulin-deficient rats. Male Wistar rats were first rendered diabetic through streptozotocin (STZ) administration and then intraperitoneally injected with AICAR for 7 consecutive days. Food and water intake, ambulatory activity, and energy expenditure were assessed at the end of the AICAR-treatment period. Blood was collected for circulating leptin measurement and the hypothalami were extracted for the determination of suppressor of cytokine signaling 3 (SOCS3) content, as well as the content and phosphorylation of AMP-kinase (AMPK), acetyl-CoA carboxylase (ACC), and the signal transducer and activator of transcription 3 (STAT3). Rats were thoroughly dissected for adiposity and lean body mass (LBM) determinations. In non-diabetic rats, despite reducing adiposity, AICAR increased (â¼1.7-fold) circulating leptin and reduced hypothalamic SOCS3 content and food intake by 67% and 25%, respectively. The anorexic effect of AICAR was lost in diabetic rats, even though hypothalamic AMPK and ACC phosphorylation markedly decreased in these animals. Importantly, hypothalamic SOCS3 and STAT3 levels remained elevated and reduced, respectively, after treatment of insulin-deficient rats with AICAR. Diabetic rats were lethargic and displayed marked losses of fat and LBM. AICAR treatment increased ambulatory activity and whole-body energy expenditure while also attenuating diabetes-induced fat and LBM losses. In conclusion, AICAR did not reverse hyperphagia, but it promoted anti-catabolic effects on skeletal muscle and fat, enhanced spontaneous physical activity, and improved the ability of rats to cope with the diabetes-induced dysfunctional alterations in glucose metabolism and whole-body energy homeostasis.
Asunto(s)
Proteínas Quinasas Activadas por AMP/metabolismo , Aminoimidazol Carboxamida/análogos & derivados , Hipotálamo/efectos de los fármacos , Hipotálamo/metabolismo , Insulina/deficiencia , Ribonucleósidos/administración & dosificación , Ribonucleósidos/farmacología , Acetil-CoA Carboxilasa/metabolismo , Tejido Adiposo/efectos de los fármacos , Tejido Adiposo/metabolismo , Aminoimidazol Carboxamida/administración & dosificación , Aminoimidazol Carboxamida/farmacología , Animales , Depresores del Apetito/administración & dosificación , Depresores del Apetito/farmacología , Composición Corporal/efectos de los fármacos , Peso Corporal/efectos de los fármacos , Diabetes Mellitus/sangre , Diabetes Mellitus/metabolismo , Diabetes Mellitus/patología , Diabetes Mellitus/fisiopatología , Ingestión de Líquidos/efectos de los fármacos , Metabolismo Energético/efectos de los fármacos , Hipotálamo/fisiopatología , Insulina/sangre , Leptina/sangre , Leptina/metabolismo , Masculino , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/metabolismo , Fosforilación/efectos de los fármacos , Ratas , Ratas Wistar , Factor de Transcripción STAT3/metabolismo , Transducción de Señal/efectos de los fármacos , Proteína 3 Supresora de la Señalización de Citocinas , Proteínas Supresoras de la Señalización de Citocinas/metabolismoRESUMEN
Rubinoboletus ballouii is an edible mushroom wildly grown in Yunnan province, China. Up till now, little was known about the chemical and biological properties of this mushroom. The aim of this study was to investigate the immunomodulatory effects of the ethanolic extract of Rubinoboletus ballouii and its fractions on human peripheral blood mononuclear cells (PBMCs) using bioactivity-guided fractionation. The crude extract of the fruiting bodies of RB was fractionated by high-speed counter current chromatography (HSCCC). Twelve fractions were obtained and the third fraction (Fraction C) exerted the most potent anti-inflammatory activities in mitogen-activated PBMCs. Further fractionation of fraction C led to the isolation of two single compounds which were elucidated as 1-ribofuranosyl-s-triazin-2(1H)-one and pistillarin, respectively. The results showed that both 1-ribofuranosyl-s-triazin-2(1H)-one and pistillarin exhibited significant immunosuppressive effects on phytohemagglutinin (PHA)-stimulated human PBMCs by inhibiting [methyl-(3)H]-thymidine uptake and inflammatory cytokines productions such as tumor necrosis factor (TNF)-α, interleukin (IL)-10, interferon (IFN)-γ and IL-1ß. Besides, 1-ribofuranosyl-s-triazin-2(1H)-one was firstly found in natural resources, and pistillarin was also isolated from the family Boletaceae for the first time. They exhibited great potential in developing as anti-inflammatory reagents.
Asunto(s)
Agaricales/química , Catecoles/aislamiento & purificación , Extractos Celulares/aislamiento & purificación , Citocinas/efectos de los fármacos , Inmunosupresores/aislamiento & purificación , Ribonucleósidos/aislamiento & purificación , Triazinas/aislamiento & purificación , Bioensayo , Catecoles/química , Catecoles/farmacología , Ciclo Celular/efectos de los fármacos , Extractos Celulares/química , Extractos Celulares/farmacología , Fraccionamiento Químico , Citocinas/metabolismo , Humanos , Inmunosupresores/química , Inmunosupresores/farmacología , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/metabolismo , Estructura Molecular , Ribonucleósidos/química , Ribonucleósidos/farmacología , Triazinas/química , Triazinas/farmacologíaRESUMEN
Maribavir (MBV), a UL97 inhibitor, shows good oral bioavailability, low host cell toxicity, and theoretical benefits to inhibit cross-resistant viruses. We herein examined clinical and virological outcomes of 12 patients, including 3 bone marrow recipients and 9 organ recipients infected with resistant cytomegalovirus (CMV) and treated with MBV during 2011-2012. All received at least 800-mg daily doses. They had developed clinical (12/12) and/or virological (11/12) resistance to CMV infection. Based on a decrease of viral load in blood >1.5 log copies/mL half of them responded to MBV treatment. The individual changes varied from a rapid decrease in viral load (n = 4) to no response (n = 3) with some late response slowly decreasing viremia (n = 3). In 2 cases MBV was used as secondary prophylaxis. No clear parameter emerged as a clinical surrogate for nonresponse to MBV. These results contrast with the lack of efficacy in phase III trials of MBV prophylaxis among stem cell recipients, which were possibly due to low doses or inadequate timing of drug initiation in the study. Additional clinical and surrogate laboratory markers are needed to determine antiviral responses to guide MBV use. Dosage ranging studies might benefit future MBV use.
Asunto(s)
Antivirales/uso terapéutico , Bencimidazoles/uso terapéutico , Infecciones por Citomegalovirus/tratamiento farmacológico , Trasplante de Órganos , Ribonucleósidos/uso terapéutico , Adulto , Antivirales/farmacología , Bencimidazoles/farmacología , Citomegalovirus/efectos de los fármacos , Francia , Genotipo , Humanos , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Fosfotransferasas (Aceptor de Grupo Alcohol)/genética , Ribonucleósidos/farmacologíaRESUMEN
Low-power laser irradiation (LPLI) has been shown to exert promotive effects on cell survival and proliferation through activation of various signaling pathways. Estrogen receptors (ERs, ERα, and ERß) are ligand-activated transcription factors, which regulate target gene expression, promote cell proliferation, and resist apoptosis. However, it is unclear whether LPLI could induce ligand-independent activation of ERs. In the present study, we investigated the subcellular pools, nuclear redistribution, and transcriptional activity of ERs under LPLI (1.2 J/cm(2), 633 nm) treatment using single-molecule fluorescence imaging and dual-luciferase reporter assay. We found that ERs were not only localized to nucleus, but also existed in mitochondria. Moreover, we found that LPLI induced nuclear redistribution and transcriptional activity of ERs in a ligand-independent manner. Our further investigation showed that PI3-K/Akt signaling cascade was involved in LPLI-induced activation of ERs. Wortmannin, a PI3-K inhibitor, or triciribine (API-2), a specific Akt inhibitor, potently suppressed the nuclear redistribution and transcriptional activity of ERs induced by LPLI, revealing that PI3-K/Akt signaling cascade was required for the activation of ERs induced by LPLI. Collectively, we demonstrated the first time that LPLI induced the ligand-independent nuclear redistribution and transcriptional activity of ERs, which were dependent on the activity of PI3-K/Akt. Our findings provide direct evidence for the molecular mechanisms of LPLI-induced transcription factor activation.
Asunto(s)
Regulación Neoplásica de la Expresión Génica/efectos de la radiación , Fosfatidilinositol 3-Quinasas , Proteínas Proto-Oncogénicas c-akt , Receptores de Estrógenos/metabolismo , Androstadienos/farmacología , Animales , Apoptosis/efectos de la radiación , Células COS , Núcleo Celular/metabolismo , Proliferación Celular/efectos de la radiación , Chlorocebus aethiops , Estrógenos/metabolismo , Células Hep G2 , Humanos , Terapia por Luz de Baja Intensidad , Células MCF-7 , Mitocondrias/metabolismo , Mitocondrias/efectos de la radiación , Fosfatidilinositol 3-Quinasas/metabolismo , Inhibidores de las Quinasa Fosfoinosítidos-3 , Proteínas Proto-Oncogénicas c-akt/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-akt/metabolismo , Especies Reactivas de Oxígeno , Ribonucleósidos/farmacología , Transducción de Señal/efectos de la radiación , WortmaninaRESUMEN
Current therapy options to treat infections with human cytomegalovirus face severe limitations leading to a continued search for novel drug candidates. Here, we describe novel characteristics of the strong antiviral potency of the drug artesunate. In vitro virus replication systems were applied to analyze a number of laboratory and clinically relevant strains of human cytomegalovirus. An inhibitory block at a very early stage of infection was demonstrated. Time-of-addition experiments indicated that the antiviral efficacy could be optimized when artesunate was applied as fractional doses consecutively added post-infection. Artesunate showed a clearly higher anti-cytomegaloviral activity than its parental drug artemisinin (approximately 10-fold) or other artesunate-related compounds. Mean IC(50) values of artesunate for a variety of standard therapy-resistant virus mutants were within a 2-fold range compared to wild-type virus. Furthermore, a synergistic effect was identified when artesunate was combined with the mechanistically distinct antiviral compound maribavir. These findings point to unique antiviral properties of artesunate which may offer an advantage over standard antiviral therapy particularly in cases of drug resistance.
Asunto(s)
Antivirales/farmacología , Artemisininas/farmacología , Citomegalovirus/efectos de los fármacos , Artesunato , Bencimidazoles/farmacología , Línea Celular , Sinergismo Farmacológico , Humanos , Concentración 50 Inhibidora , Pruebas de Sensibilidad Microbiana , Ribonucleósidos/farmacología , Replicación Viral/efectos de los fármacosRESUMEN
BACKGROUND: Despite advances in cytomegalovirus (CMV) prophylaxis and therapy, some transplant recipients still develop refractory CMV infections. Maribavir (MBV), an investigational benzimidazole antiviral agent, acts by a mechanism different from that of existing anti-CMV drugs. Previous Phase I and II studies have demonstrated a favorable safety profile for MBV, but its utility in treatment of complex CMV syndromes is unknown. METHODS: Between June and December 2008, MBV was released for use under individual emergency investigational new drug applications requested by treating physicians and approved by the US Food and Drug Administration and local institutional review boards. Six patients (5 solid organ transplant recipients and 1 hematopoietic stem cell transplant recipient) who had failed to respond to other therapies and/or had known ganciclovir-resistant CMV were treated with MBV at a starting oral dose of 400 mg twice daily. RESULTS: Patients were treated for a median of 207 days (range, 15-376). Four of 6 patients had no detectable CMV DNAemia within 6 weeks of starting MBV therapy. One patient, who had an initial viral load of 1.8 million copies/mL, developed MBV resistance mutations. One patient, who had low serum levels of MBV, had persistent CMV DNAemia and viruria without developing genotypic or phenotypic resistance to MBV. One patient cleared CMV DNAemia, but died of pneumonia and multiorgan failure. No significant adverse effects attributable to MBV were observed. CONCLUSIONS: MBV deserves further systematic evaluation as treatment for CMV infection that is resistant and/or refractory to standard therapies, but its optimal dose, duration of therapy, and use in combinations versus as a single agent have yet to be determined.
Asunto(s)
Antivirales/administración & dosificación , Bencimidazoles/administración & dosificación , Infecciones por Citomegalovirus/tratamiento farmacológico , Farmacorresistencia Viral , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Trasplante de Órganos/efectos adversos , Ribonucleósidos/administración & dosificación , Adolescente , Adulto , Anciano , Antivirales/efectos adversos , Antivirales/farmacología , Antivirales/uso terapéutico , Bencimidazoles/efectos adversos , Bencimidazoles/farmacología , Bencimidazoles/uso terapéutico , Citomegalovirus/efectos de los fármacos , Citomegalovirus/genética , Citomegalovirus/aislamiento & purificación , Infecciones por Citomegalovirus/virología , Femenino , Ganciclovir/farmacología , Humanos , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Ribonucleósidos/efectos adversos , Ribonucleósidos/farmacología , Ribonucleósidos/uso terapéutico , Resultado del TratamientoRESUMEN
Acadesine is an adenosine receptor agonist (ARA) in development for the treatment of ischaemia-reperfusion injury and chronic lymphocytic leukaemia. Schering-Plough is developing the compound as a cardioprotective agent in ischaemia-reperfusion injury. Avancell and Protherics are co-developing acadesine for the treatment of B-cell chronic lymphocytic leukaemia (B-CLL). Clinical development is underway for both indications. In January 2005, PeriCor Therapeutics obtained a sublicense for the worldwide rights to acadesine and three additional ARA compounds from Dr Mangano who was a founder of the company. Dr Mangano acquired the license of worldwide rights for acadesine from Metabasis in November 2000.Previously, in December 1997, all intellectual property rights and data covering acadesine and related ARAs for cardiovascular and cerebrovascular disorders were transferred from the original developer, Gensia Sicor (SICOR), to Metabasis Therapeutics in an asset-liability transfer agreement.Schering-Plough intends to conduct a randomized, placebo-controlled phase III trial, which is required for regulatory approval. The trial will further evaluate acadesine in patients deemed to be at high risk of ischaemic heart disorders. Patients will have either undergone coronary artery bypass graft surgery, or have a history of cardiovascular events (heart attack or stroke). Protherics presented 2-year mortality results from a study of acadesine, investigating long-term mortality after perioperative myocardial infarction, at the 55th Scientific Session of the American College of Cardiology in 2006.Previously, Gensia Sicor conducted clinical trials of acadesine in Canada, the EU and the US; results of these trials were inconclusive. Acadesine has been shown to selectively cause the death of B-cells, whilst sparing T-cells when administered to blood samples taken from patients with B-CLL. Orphan drug status has been granted for acadesine in the EU for the treatment of B-cell chronic lymphocytic leukaemia.
Asunto(s)
Aminoimidazol Carboxamida/análogos & derivados , Agonistas del Receptor Purinérgico P1 , Ribonucleósidos/farmacología , Aminoimidazol Carboxamida/efectos adversos , Aminoimidazol Carboxamida/farmacocinética , Aminoimidazol Carboxamida/farmacología , Animales , Ensayos Clínicos como Asunto , Diseño de Fármacos , Evaluación Preclínica de Medicamentos , Interacciones Farmacológicas , Humanos , Ribonucleósidos/efectos adversos , Ribonucleósidos/farmacocinéticaRESUMEN
The objectives were: (1) to test the association of methotrexate (MTX) efficacy in rat adjuvant arthritis (rat AA) with interference of purine biosynthesis and adenosine metabolism and (2) to test the efficacy of erythro-9-(2-hydroxynon-3-yl) adenine (EHNA), an inhibitor of adenosine deaminase, and the efficacy of aminoimidazolecarboxamide (AICA) riboside plus MTX in rat AA. Radiographic and histologic examinations of the hind limbs were measures of efficacy. Urinary excretions of AICA and adenosine were markers of AICA ribotide transformylase inhibition (i.e., blockage of purine biosynthesis) and interference with adenosine metabolism, respectively. AICA and adenosine excretions increased during the day of MTX dosing (treatment day) compared to the previous baseline day in animals responding well to MTX (i.e., low radiographic and histologic scores). Based on radiographic and histologic scores, adjuvant injected rats were separated into two disease categories (i.e., no/mild and moderate/severe). Only AICA excretion was significantly elevated on the treatment day in rat AA with no/mild disease (i.e., those responding well to MTX therapy). AICA (not adenosine) excretion was significantly correlated with the above scores. EHNA was not efficacious, even at toxic levels, while AICA riboside potentiated the efficacy of MTX. The data suggests that efficacious MTX therapy in rat AA (1) blocks purine biosynthesis; (2) increases in in vivo AICA levels. Also adenosine accumulation and blockage of adenosine deaminase (i.e., by EHNA) appear to be less critical to MTX efficacy. Increased levels of AICA metabolites may suppress the immune response in rat AA.
Asunto(s)
Adenina/análogos & derivados , Aminoimidazol Carboxamida/análogos & derivados , Antirreumáticos/farmacología , Artritis Experimental/tratamiento farmacológico , Inhibidores Enzimáticos/farmacología , Metotrexato/farmacología , Purinas/metabolismo , Ribonucleósidos/farmacología , Adenina/farmacología , Adenina/uso terapéutico , Adenosina/orina , Adenosina Desaminasa/metabolismo , Inhibidores de la Adenosina Desaminasa , Aminoimidazol Carboxamida/farmacología , Aminoimidazol Carboxamida/uso terapéutico , Aminoimidazol Carboxamida/orina , Animales , Antirreumáticos/uso terapéutico , Artritis Experimental/enzimología , Artritis Experimental/patología , Artritis Experimental/orina , Biomarcadores/orina , Relación Dosis-Respuesta a Droga , Quimioterapia Combinada , Inhibidores Enzimáticos/uso terapéutico , Metotrexato/uso terapéutico , Fosforribosilaminoimidazolcarboxamida-Formiltransferasa/antagonistas & inhibidores , Fosforribosilaminoimidazolcarboxamida-Formiltransferasa/metabolismo , Purinas/orina , Ratas , Ribonucleósidos/uso terapéuticoRESUMEN
The long-term treatment of herpesvirus infections with current antivirals leads to the development of drug-resistant viruses. Because currently available antivirals finally target the viral DNA polymerase, mutant resistant to one drug often shows cross-resistance to other drugs. This evidence highlights the need for the development of new antivirals that have the different viral targets. Recently, high-through-put screening of large compound collections for inhibiting specific viral enzymes, or in vitro cell culture assay, has identified several new antivirals. These include the inhibitors of helicase/primase complex, terminase complex, portal protein and UL97 protein kinase. This review will focus on these new compounds that directly inhibit viral replication.
Asunto(s)
Antivirales , Infecciones por Herpesviridae/tratamiento farmacológico , Herpesviridae , Animales , Antivirales/farmacología , Bencimidazoles/farmacología , Bencimidazoles/uso terapéutico , ADN Helicasas/antagonistas & inhibidores , Diseño de Fármacos , Evaluación Preclínica de Medicamentos , Farmacorresistencia Viral , Endodesoxirribonucleasas/antagonistas & inhibidores , Herpesviridae/efectos de los fármacos , Herpesviridae/enzimología , Infecciones por Herpesviridae/virología , Humanos , Péptidos y Proteínas de Señalización Intracelular/farmacología , Péptidos y Proteínas de Señalización Intracelular/uso terapéutico , Inhibidores de la Síntesis del Ácido Nucleico , Ribonucleósidos/farmacología , Ribonucleósidos/uso terapéuticoRESUMEN
Synthesis, conformational analysis and antitumor evaluation of 2'- and 3'-C-methyl analogues of mizoribine (bredinine, 4-carbamoyl-1-beta-D-ribofuranosylimidazole-5-olate) are reported.
Asunto(s)
Antibióticos Antineoplásicos/síntesis química , Inhibidores Enzimáticos/síntesis química , IMP Deshidrogenasa/antagonistas & inhibidores , Ribonucleósidos/síntesis química , Antibióticos Antineoplásicos/farmacología , Evaluación Preclínica de Medicamentos , Inhibidores Enzimáticos/farmacocinética , Humanos , Células K562 , Estructura Molecular , Ribonucleósidos/farmacologíaRESUMEN
The AMP-activated kinase has been proposed to be an important intracellular energy sensor because the enzyme controls lipid and glucose oxidation. In the corresponding knockout mice, insulin-stimulated muscle glycogen synthesis and glucose tolerance are reduced. In addition, these mice excrete catecholamines in excess, suggesting that the central and autonomic nervous systems are impaired. Indeed, in the brain, fuel sensor mechanisms have been described, and recently, evidence has shown that the AMP-activated kinase could control food intake. We show in this study that the intracerebroventricular infusion of 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside (AICAR), a pharmacological AMP-activated kinase activator, increased insulin-stimulated muscle glycogen synthesis and insulin sensitivity during a hyperinsulinemic clamp. Similarly, we infused AICAR in the brain of fasted mice, i.e. when insulinemia was low, and showed that muscle glycogen synthesis was also increased. We then studied the effect of a cerebral infusion of the peripheral signals, i.e. insulin and glucose, known to be detected by the brain. The cerebral infusion of insulin increased muscle glycogen synthesis. This effect was blunted by the coinfusion of glucose, which induced insulin resistance. Importantly, the cerebral injections of AICAR, insulin, and glucose were associated with variations in the phosphorylation state of the AMP-activated kinase in the hypothalamus. In conclusion, our data showed for the first time that 1) the brain is sensitive to insulin and glucose for the regulation of muscle glycogen synthesis; and 2) the cerebral infusion of AICAR enhances insulin sensitivity. Although the above mechanisms are correlated with the regulation of AMP-activated kinase, the direct involvement of the enzyme in the mechanism remains to be demonstrated.
Asunto(s)
Aminoimidazol Carboxamida/análogos & derivados , Aminoimidazol Carboxamida/farmacología , Glucosa/farmacocinética , Hipoglucemiantes/farmacología , Insulina/farmacología , Complejos Multienzimáticos/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Ribonucleósidos/farmacología , Proteínas Quinasas Activadas por AMP , Animales , Ayuno/fisiología , Glucógeno/biosíntesis , Hiperinsulinismo/metabolismo , Hipotálamo/efectos de los fármacos , Hipotálamo/enzimología , Inyecciones Intraventriculares , Masculino , Ratones , Ratones Endogámicos C57BL , Músculo Esquelético/metabolismo , Fosforilación/efectos de los fármacosRESUMEN
AMP-activated protein kinase (AMPK) is the downstream component of a protein kinase cascade that acts as an intracellular energy sensor maintaining the energy balance within the cell. The finding that leptin and adiponectin activate AMPK to alter metabolic pathways in muscle and liver provides direct evidence for this role in peripheral tissues. The hypothalamus is a key regulator of food intake and energy balance, coordinating body adiposity and nutritional state in response to peripheral hormones, such as leptin, peptide YY-(3-36), and ghrelin. To date the hormonal regulation of AMPK in the hypothalamus, or its potential role in the control of food intake, have not been reported. Here we demonstrate that counter-regulatory hormones involved in appetite control regulate AMPK activity and that pharmacological activation of AMPK in the hypothalamus increases food intake. In vivo administration of leptin, which leads to a reduction in food intake, decreases hypothalamic AMPK activity. By contrast, injection of ghrelin in vivo, which increases food intake, stimulates AMPK activity in the hypothalamus. Consistent with the effect of ghrelin, injection of 5-amino-4-imidazole carboxamide riboside, a pharmacological activator of AMPK, into either the third cerebral ventricle or directly into the paraventricular nucleus of the hypothalamus significantly increased food intake. These results suggest that AMPK is regulated in the hypothalamus by hormones which regulate food intake. Furthermore, direct pharmacological activation of AMPK in the hypothalamus is sufficient to increase food intake. These findings demonstrate that AMPK plays a role in the regulation of feeding and identify AMPK as a novel target for anti-obesity drugs.
Asunto(s)
Aminoimidazol Carboxamida/análogos & derivados , Ingestión de Alimentos , Péptidos y Proteínas de Señalización Intercelular , Complejos Multienzimáticos/fisiología , Proteínas Serina-Treonina Quinasas/fisiología , Proteínas Quinasas Activadas por AMP , Adiponectina , Aminoimidazol Carboxamida/farmacología , Animales , Regulación del Apetito , Western Blotting , Encéfalo/metabolismo , Ghrelina , Hipotálamo/metabolismo , Leptina/metabolismo , Hígado/metabolismo , Músculos/metabolismo , Hormonas Peptídicas/metabolismo , Péptido YY/farmacología , Proteínas/metabolismo , Ratas , Ratas Wistar , Ribonucleósidos/farmacología , Factores de TiempoRESUMEN
Different arabinosides and ribosides, viz. Ara-DDA or 9(1-beta-D-arabinofuranosyl) 1,3-dideazaadenine (6), Ara-NDDP or 9(1-beta-D-arabinofuranosyl) 4-nitro-1,3-dideazapurine (7), Ara-DKP or 1(1-beta-D-arabinofuranosyl) diketopiperazine (8), Ribo-DDA or 9(1-beta-D-ribofuranosyl) 1,3-dideazaadenine (9) and Ribo-NDDP or 9(1-beta-D-ribofuranosyl) 4-nitro-1,3-dideazapurine (10) have been synthesized as probable antiviral agents. The arabinosides have been synthesized using the catalyst TDA-1 that causes stereospecific formation of beta-nucleosides while a one-pot synthesis procedure was adopted for the synthesis of the ribonucleosides where beta-anomers were obtained in higher yields. All the five nucleoside analogs have been screened for antiviral property against HIV-1 (IIIB), HSV-1 and 2, parainfluenza-3, reovirus-1 and many others. It was observed that arabinosides had greater inhibitory action than ribosides. The compound 7 or Ara-NDDP has shown maximum inhibition of HIV-1 replication than the rest of the molecules with an IC50 of 79.4 microg/mL.