RESUMEN
Objective: Roxadustat is a new medication for the treatment of renal anemia. EPO (erythropoietin)-the current treatment standard-has been reported to enhance platelet activation and production. However, to date, the effect of roxadustat on platelets is unclear. To address this deficiency, herein, we have evaluated the effect of roxadustat on platelet production and function. Approach and Results: We performed several mouse platelet functional assays in the presence/absence of in vitro and in vivo roxadustat treatment. Both healthy and 5/6 nephrectomized mice were utilized. The effect of roxadustat on platelet function of healthy volunteers and chronic kidney disease patients was also evaluated. For platelet production, megakaryocyte maturation and proplatelet formation were assayed in vitro. Peripheral platelet and bone marrow megakaryocyte counts were also determined. We found that roxadustat could not stimulate washed platelets directly, and platelet aggregation, spreading, clot retraction, and P-selectin/JON/A exposure were similar with or without in vitro or in vivo roxadustat treatment among both healthy and 5/6 nephrectomized mice. In vivo mouse thrombosis models were additionally performed, and no differences were detected between the vehicle and roxadustat treatment groups. EPO, which was considered a positive control in the present study, promoted platelet function and production as reported previously. Megakaryocyte maturation and proplatelet formation were also not significantly different between control mice and those treated with roxadustat. After receiving roxadustat for 14 days, no difference in the peripheral platelet count was observed in the mice. Conclusions: Administration of roxadustat has no significant impact on platelet production and function.
Asunto(s)
Coagulación Sanguínea/efectos de los fármacos , Plaquetas/efectos de los fármacos , Eritropoyetina/farmacología , Glicina/análogos & derivados , Hematínicos/farmacología , Isoquinolinas/farmacología , Activación Plaquetaria/efectos de los fármacos , Trombopoyesis/efectos de los fármacos , Trombosis/sangre , Animales , Plaquetas/metabolismo , Estudios de Casos y Controles , Modelos Animales de Enfermedad , Eritropoyetina/toxicidad , Glicina/farmacología , Glicina/toxicidad , Hematínicos/toxicidad , Humanos , Isoquinolinas/toxicidad , Masculino , Ratones Endogámicos C57BL , Insuficiencia Renal Crónica/sangre , Insuficiencia Renal Crónica/etiología , Trombosis/etiologíaRESUMEN
Considerable data have suggested that acute kidney injury (AKI) is often incompletely repaired and could lead to chronic kidney disease (CKD). As we known, toxin-induced nephropathy triggers the rapid production of proinflammatory mediators and the prolonged inflammation allows the injured kidneys to develop interstitial fibrosis. In our previous study, fatty acid-binding protein 4 (Fabp4) has been reported to be involved in the process of AKI. However, whether Fabp4 plays crucial roles in toxin-induced kidney injury remained unclear. To explore the effect and mechanism of Fabp4 on toxin induced kidney injury, folic acid (FA) and aristolochic acid (AA) animal models were used. Both FA and AA injected mice developed severe renal dysfunction and dramatically inflammatory response (IL-6, MCP1 and TNF-a), which further lead to early fibrosis confirmed by the accumulation of extracellular matrix proteins (α-Sma, Fn, Col1 and Col4). Importantly, we found that FA and AA induced-kidney injury triggered the high expression of Fabp4 mRNA/protein in tubular epithelial cells. Furthermore, pharmacological and genetic inhibition of Fabp4 significantly attenuated FA and AA induced renal dysfunction, pathological damage, and early fibrosis via the regulation of inflammation, which is mediated by suppressing p-p65/p-stat3 expression via enhancing Pparγ activity. In summary, Fabp4 in tubular epithelial cells exerted the deleterious effects during the recovery of FA and AA induced kidney injury and the inhibition of Fabp4 might be an effective therapeutic strategy against the progressive AKI.
Asunto(s)
Lesión Renal Aguda/tratamiento farmacológico , Compuestos de Bifenilo/farmacología , Proteínas de Unión a Ácidos Grasos/antagonistas & inhibidores , Fibrosis/prevención & control , Inflamación/tratamiento farmacológico , Pirazoles/farmacología , Lesión Renal Aguda/inducido químicamente , Lesión Renal Aguda/inmunología , Lesión Renal Aguda/metabolismo , Animales , Ácidos Aristolóquicos/toxicidad , Carcinógenos/toxicidad , Modelos Animales de Enfermedad , Proteínas de Unión a Ácidos Grasos/genética , Proteínas de Unión a Ácidos Grasos/metabolismo , Fibrosis/inducido químicamente , Fibrosis/inmunología , Fibrosis/metabolismo , Ácido Fólico/toxicidad , Hematínicos/toxicidad , Inflamación/inmunología , Inflamación/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , ARN Interferente Pequeño/genéticaRESUMEN
OBJECTIVE: Iron is emerging as a key player in aging-associated diseases due to its propensity for driving free radical formation. Studies examining the role of iron in the pathogenesis of primary osteoarthritis (OA) are limited. Our objective was to establish a direct relationship between excess iron and OA by administering iron dextran to a guinea pig strain with decreased propensity for developing this disease. DESIGN: Twenty, 12-week-old Strain 13 guinea pigs received either iron dextran or dextran control intraperitoneally once weekly for 4 weeks; termination occurred at 16 weeks of age. Iron levels were determined systemically (serum and liver) and within diarthrodial joints [femoral head articular cartilage and infrapatellar fat pads (IFPs) of knee joints]. One knee was collected to score structural changes associated with OA via microcomputed tomography (microCT) and histology using published grading schemes. Articular cartilage and IFPs were harvested from contralateral knees for gene expression analyses. RESULTS: Iron overload was confirmed systemically via increased serum iron and liver iron concentration. Articular cartilage and IFPs in the iron dextran group also had higher levels of iron. Excess iron worsened knee OA using both microCT and histologic scoring systems. Gene analyses revealed that exogenous iron altered the expression of iron trafficking proteins, select cytokines, and structural components of cartilage. CONCLUSION: These results demonstrate that systemic iron overload caused cellular iron accumulation in the knee joint. This excess iron is associated with increased expression of local inflammatory mediators and early onset and progression of knee joint OA in Strain 13 animals.
Asunto(s)
Tejido Adiposo/metabolismo , Cartílago Articular/metabolismo , Sobrecarga de Hierro/fisiopatología , Osteoartritis/fisiopatología , Agrecanos/genética , Animales , Antígenos CD/genética , Antígenos de Diferenciación Mielomonocítica/genética , Apoferritinas/genética , Proteínas de Transporte de Catión/genética , Colágeno Tipo II/genética , Femenino , Expresión Génica , Cobayas , Hematínicos/toxicidad , Interleucina-1beta/genética , Interleucina-6/genética , Sobrecarga de Hierro/inducido químicamente , Sobrecarga de Hierro/metabolismo , Sobrecarga de Hierro/patología , Complejo Hierro-Dextran/toxicidad , Articulación de la Rodilla/diagnóstico por imagen , Articulación de la Rodilla/metabolismo , Articulación de la Rodilla/patología , Hígado/metabolismo , Masculino , Osteoartritis/diagnóstico por imagen , Osteoartritis/metabolismo , Osteoartritis/patología , Osteoartritis de la Rodilla/diagnóstico por imagen , Osteoartritis de la Rodilla/metabolismo , Osteoartritis de la Rodilla/patología , Osteoartritis de la Rodilla/fisiopatología , ARN Mensajero/metabolismo , Receptores de Superficie Celular/genética , Receptores de Transferrina/genética , Espectrofotometría Atómica , Factor de Crecimiento Transformador beta1/genética , Factor de Necrosis Tumoral alfa/genética , Microtomografía por Rayos XRESUMEN
The aim of the study was to examine the reproducibility of a rat model to assess the preclinical similarity in safety profiles and tissue accumulation of iron products. Accordingly, the effect of several doses of intravenously administered Venofer® and of Ferrlecit® on blood parameters, and on kidney and particularly liver toxicity were examined in non-anemic Sprague Dawley rats. The different analysis showed neither a clear treatment nor a dose effect after multiple injections. The parameters measured in this rat strain showed some iron induced adverse effects, but these could not be correlated to treatment specific differences. The findings presented in this paper indicate the difficulty to define a useful preclinical model to evaluate iron-based nano-colloidal preparations.
Asunto(s)
Hematínicos/toxicidad , Riñón/efectos de los fármacos , Hígado/efectos de los fármacos , Modelos Animales , Ratas , Animales , Coloides/administración & dosificación , Coloides/toxicidad , Compuestos Férricos/administración & dosificación , Compuestos Férricos/toxicidad , Sacarato de Óxido Férrico , Ácido Glucárico/administración & dosificación , Ácido Glucárico/toxicidad , Hematínicos/administración & dosificación , Infusiones Intravenosas , Inyecciones Intravenosas , Masculino , Nanopartículas/administración & dosificación , Nanopartículas/toxicidad , Ratas Sprague-Dawley , Reproducibilidad de los ResultadosRESUMEN
CONTEXT: Angelica sinensis L. (Umbelliferae) has medicinal properties. OBJECTIVES: The present study evaluates the haematopoietic effects of A. sinensis polysaccharides (ASP) against lisinopril-induced anaemia. MATERIALS AND METHODS: Thirty healthy adult male albino rats were randomly divided into five groups (n = 6). Group I was control group. Group II was treated with angiotensin-converting enzyme inhibitor (ACEI, 20 mg/kg/day) to induce anaemia. In group III, erythropoietin (EPO, 100 IU/kg/each) was administered in combination with ACEI. Group IV was treated with ASP (1 g/kg/day), extracted from A. sinensis root caps. In Group V, ASP (1 g/kg/day) was treated with ACEI. After 28 days, blood and tissue samples were collected for haematological and histopathological analysis, respectively. RESULTS: The results showed that ACEI significantly reduced the haemoglobin (Hb, 10.0 g/dL), packed cell volume (PCV, 39.5%), red blood cells (RBCs, 6.2 million/mm3), mean corpuscular volume (MCV, 53.5 fL) and mean corpuscular haemoglobin (MCH, 16.2 pg/cell) values. In the group treated with ASP, the Hb (13.7 g/dL) and RBCs (7.8 million/mm3) increased significantly (p < 0.05). The combination of ASP and ACEI led to the significant (p < 0.05) reduction in Hb (10.7 g/dL), PCV (33.3%), RBCs (6.0 million/mm3), MCV (54.42 fL) and MCH (16.44 pg/cell) values. While histopathological examination of the liver and kidney cells showed a mild degree of toxicity in the ASP-treated group. CONCLUSION: ASP has a potentiating effect on haematological parameters when given alone. However, when administered simultaneously with lisinopril, it showed an unfavourable effect with more complicated anaemia so it should not be used with ACEIs.
Asunto(s)
Anemia/tratamiento farmacológico , Angelica sinensis/química , Eritrocitos/efectos de los fármacos , Hematínicos/farmacología , Hematopoyesis/efectos de los fármacos , Lisinopril , Extractos Vegetales/farmacología , Cápsula de Raíz de Planta/química , Polisacáridos/farmacología , Anemia/sangre , Anemia/inducido químicamente , Animales , Biomarcadores/sangre , Modelos Animales de Enfermedad , Índices de Eritrocitos , Eritrocitos/metabolismo , Eritropoyetina/farmacología , Hematínicos/aislamiento & purificación , Hematínicos/toxicidad , Hematócrito , Hemoglobinas/metabolismo , Interacciones de Hierba-Droga , Masculino , Fitoterapia , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/toxicidad , Plantas Medicinales , Polisacáridos/aislamiento & purificación , Polisacáridos/toxicidad , Ratas Wistar , Factores de TiempoRESUMEN
EPO-018B, a synthetic peptide-based erythropoiesis stimulating agent (ESA), is mainly designed for treatment of anemia caused by chronic renal failure and chemotherapy against cancer. It overcomes the deficiencies of currently approved ESA, including the frequent administration of temperature-sensitive recombinant protein and anti-EPO antibody-mediated pure red cell aplasia (PRCA). This study was designed to evaluate the potential chronic toxicity of EPO-018B. Subcutaneous administration doses were designed as 0, 0.2, 1 and 10mg/kg for six months for 160 rats (20/gender/group) and 0, 0.3, 3 and 20mg/kg for nine months for 32 monkeys (4/gender/group) once every three weeks. The vehicles received the same volume of physiological saline injection. All animals survived to the scheduled necropsies after six weeks (for rats) and fourteen weeks (for monkeys) recovery period, except for the two high-dose female rats and two high-dose male monkeys, which were considered related to the increased RBCs, chronic blood hyperviscosity and chronic cardiac injury. EPO-018B is supposed to be subcutaneously injected once every month and the intended human therapeutic dose is 0.025mg/kg. The study findings at 0.2mg/kg for rats and 0.3mg/kg for monkeys were considered to be the study NOAEL (the no observed adverse effect level), which were more than ten times the intended human therapeutic dose. Higher doses caused adverse effects related to the liver toxicity, cardiotoxicity, appearance of neutralizing antibodies of EPO-018B and the decrease of serum glucose and cholesterol. Most treatment-induced effects were reversible or revealed ongoing recovery upon the discontinuation of treatment. The sequelae occurred in rats and monkeys were considered secondary to exaggerated pharmacology and would less likely occur in the intended patient population. As to the differences between human beings and animals, the safety of EPO-018B need to be further confirmed in the future clinical studies.
Asunto(s)
Eritropoyetina/análogos & derivados , Eritropoyetina/toxicidad , Hematínicos/toxicidad , Animales , Anticuerpos/sangre , Evaluación Preclínica de Medicamentos , Femenino , Hematínicos/inmunología , Pruebas Hematológicas , Macaca fascicularis , Masculino , Ratas Sprague-DawleyRESUMEN
The erythropoietin analog peginesatide was withdrawn from marketing due to unexpected severe anaphylactic reactions associated with administration of the multi-use formulation. The adverse events occurred rapidly following the first ever administration of the drug with most affected patients becoming symptomatic in less than 30min. This is most consistent with an anaphylactoid reaction due to direct activation of mast cells. Laboratory evaluation was undertaken using rat peritoneal mast cells as the model system. Initial studies showed that high concentrations of the formulated drug as well as formulated vehicle alone could cause mast cell degranulation as measured by histamine release. The purified active drug was not able to cause histamine release whereas the vehicle filtrate and lab created drug vehicle were equally potent at causing histamine release. Individual formulations of vehicle leaving one component out showed that histamine release was due to phenol. Dose response studies with phenol showed a very sharp dose response curve that was similar in three buffer systems. Cellular analysis by flow cytometry showed that the histamine release was not due to cell death, and that changes in light scatter parameters consistent with degranulation were rapidly observed. Limited testing with primary human mast cells showed a similar dose response of histamine release with exposure to phenol. To provide in vivo confirmation, rats were injected with vehicle formulated with various concentrations of phenol via a jugular vein cannula. Significant release of histamine was detected in blood samples taken 2min after dosing at the highest concentrations tested.
Asunto(s)
Degranulación de la Célula/efectos de los fármacos , Excipientes/toxicidad , Hematínicos/toxicidad , Histamina/metabolismo , Mastocitos/efectos de los fármacos , Péptidos/toxicidad , Fenol/toxicidad , Animales , Células Cultivadas , Química Farmacéutica , Relación Dosis-Respuesta a Droga , Excipientes/administración & dosificación , Excipientes/química , Femenino , Hematínicos/química , Histamina/sangre , Humanos , Inyecciones Intravenosas , Mastocitos/metabolismo , Ratones Endogámicos NOD , Péptidos/química , Fenol/administración & dosificación , Fenol/química , Cultivo Primario de Células , Ratas Sprague-Dawley , Medición de Riesgo , Factores de TiempoRESUMEN
BACKGROUND AND AIMS: Iron deficiency is common in inflammatory bowel disease, yet oral iron therapy may worsen the disease symptoms and increase systemic and local oxidative stress. The aim of this study was to compare the effects of oral ferrous sulfate and iron polymaltose complex on inflammatory and oxidative stress markers in colitic rats. METHODS: Animals were divided into four groups with ten animals each. Rats of three groups received dextran sodium sulfate to induce colitis and animals of two of these groups received 5 mg iron/kg of body weight a day, as ferrous sulfate or iron polymaltose complex, for 7 days. Gross colon anatomy, histology of colon and liver, stainings of L-ferritin, Prussian blue, hepcidin, tumor necrosis factor-α, and interleukin-6, as well serum levels of liver enzymes, inflammatory markers, and iron markers, were assessed. RESULTS: Body weight, gross anatomy, crypt injury and inflammation scores, inflammatory parameters in liver and colon, as well as serum and liver hepcidin levels were not significantly different between colitic animals without iron treatment and colitic animals treated with iron polymaltose complex. In contrast, ferrous sulfate treatment caused significant worsening of these parameters. As opposed to ferrous sulfate, iron polymaltose complex caused less or no additional oxidative stress in the colon and liver compared to colitic animals without iron treatment. CONCLUSION: Iron polymaltose complex had negligible effects on colonic tissue erosion, local or systemic oxidative stress, and local or systemic inflammation, even at high therapeutic doses, and may thus represent a valuable oral treatment of iron deficiency in inflammatory bowel disease.
Asunto(s)
Colitis/inducido químicamente , Compuestos Férricos/toxicidad , Compuestos Ferrosos/toxicidad , Estrés Oxidativo/efectos de los fármacos , Administración Oral , Animales , Colitis/patología , Sulfato de Dextran/toxicidad , Modelos Animales de Enfermedad , Femenino , Compuestos Férricos/administración & dosificación , Compuestos Ferrosos/administración & dosificación , Hematínicos/administración & dosificación , Hematínicos/toxicidad , Inflamación/inducido químicamente , Inflamación/patología , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
This study examined the safety, pharmacodynamic (PD), and pharmacokinetic (PK) biosimilarity of the human recombinant erythropoietin (EPO) products ior(®) EPOCIM and Eprex(®) following a 28-day repeated intravenous dose administration in male and female Sprague-Dawley rats with a 14-day recovery period. Safety profiling was based on clinical observations, clinical pathology, and pathology findings for control rats dosed with vehicle and rats dosed either with 30, 300, and 600 I.U./kg of ior(®) EPOCIM or 600 I.U. of Eprex(®) . Adverse findings for both ior(®) EPOCIM and Eprex(®) were similar and were a consequence of thrombotic events (ulcerative skin lesions, swollen hock joints/lameness, stomach ulcers) and decreased body weight gains, all known adverse reactions to this class of drug in rats. With the exception of stomach ulcers, all other adverse findings were fully reversible. Neither drug stimulated the production of antidrug antibodies. As expected, ior(®) EPOCIM and Eprex(®) both increased reticulocyte, red blood cell, hemoglobin, and hematocrit levels in rats. The PK of EPO following dosing with ior(®) EPOCIM was well behaved and consistent with the literature. The results of this study imply that ior(®) EPOCIM and Eprex(®) had safety profiles, PD responses, and toxicokinetic profiles that were biosimilar.
Asunto(s)
Biosimilares Farmacéuticos/farmacocinética , Biosimilares Farmacéuticos/toxicidad , Eritropoyetina/farmacocinética , Eritropoyetina/toxicidad , Hematínicos/farmacocinética , Hematínicos/toxicidad , Administración Intravenosa , Animales , Biosimilares Farmacéuticos/administración & dosificación , Biosimilares Farmacéuticos/sangre , Biosimilares Farmacéuticos/química , Química Farmacéutica , Epoetina alfa , Eritropoyetina/administración & dosificación , Eritropoyetina/sangre , Eritropoyetina/química , Femenino , Hematínicos/administración & dosificación , Hematínicos/sangre , Hematínicos/química , Masculino , Ratas Sprague-Dawley , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/sangre , Proteínas Recombinantes/química , Proteínas Recombinantes/farmacocinética , Proteínas Recombinantes/toxicidad , Medición de Riesgo , Equivalencia Terapéutica , Trombosis/inducido químicamente , Toxicocinética , Aumento de Peso/efectos de los fármacosRESUMEN
We conducted a retrospective analysis of publicly available preclinical toxicology studies with erythropoiesis-stimulating agents (ESAs) to examine common adverse events in rats, Beagle dogs, and cynomolgus monkeys. Mortality and/or thrombotic events were reported sporadically in a subset of studies and attributed to the high hematocrit (HCT) achieved in the animals. However, similarly high HCT was achieved in both high-dose and low-dose groups, but there were no reported adverse events in the low-dose group suggesting HCT was not the sole contributing factor leading to toxicity. Our analysis indicated that increased dose, dose frequency, and dosing duration in addition to high HCT contributed to mortality and thrombosis. To further evaluate this relationship, the incidence of toxicities was compared in rats administered an experimental hyperglycosylated analog of recombinant human erythropoietin (AMG 114) at varying dosing schedules in 1-month toxicity studies. The incidence of mortality and thrombotic events increased in higher dose groups and when dosed more frequently, despite a similarly high HCT in all animals. The results from the investigative study and retrospective analysis demonstrate that ESA-related toxicities in preclinical species are associated with dose level, dose frequency, and dosing duration, and not solely dependent upon a high HCT.
Asunto(s)
Hematínicos , Hematócrito , Trombosis/inducido químicamente , Animales , Investigación Biomédica , Perros , Eritropoyesis/efectos de los fármacos , Eritropoyetina/administración & dosificación , Eritropoyetina/efectos adversos , Eritropoyetina/toxicidad , Enfermedades de las Válvulas Cardíacas , Hematínicos/administración & dosificación , Hematínicos/efectos adversos , Hematínicos/toxicidad , Humanos , Macaca fascicularis , Ratas , Ratas Sprague-Dawley , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/efectos adversos , Proteínas Recombinantes/toxicidad , Estudios Retrospectivos , Neoplasias GástricasRESUMEN
Acute kidney injury (AKI) is a disease with mitochondrial dysfunction and a newly established risk factor for the development of chronic kidney disease (CKD) and fibrosis. We examined mitochondrial homeostasis in the folic acid (FA)-induced AKI model that develops early fibrosis over a rapid time course. Mice given a single dose of FA had elevated serum creatinine (3-fold) and urine glucose (2.2-fold) 1 and 2 d after injection that resolved by 4d. In contrast, peroxisome proliferator gamma coactivator 1α (PGC-1α) and mitochondrial transcription factor A (TFAM), critical transcriptional regulators of mitochondrial biogenesis (MB), were down-regulated â¼80% 1d after FA injection and remained depressed through 14 d. Multiple electron transport chain and ATP synthesis genes were also down-regulated from 1 to 14 d after FA, including NADH dehydrogenase (ubiquinone) 1 beta subcomplex 8 (NDUFß8), ATP synthase subunit ß (ATPS-ß), and cytochrome C oxidase subunit I (COXI). Mitochondrial DNA copy number was reduced â¼50% from 2 to 14 d after FA injection. Protein levels of early fibrosis markers α-smooth muscle actin and transforming growth factor ß1 were elevated at 6 and 14 d after FA. Picrosirius red staining and collagen 1A2 (COL1A2) IHC revealed staining for mature collagen deposition at 14 d. We propose that mitochondrial dysfunction induced by AKI is a persistent cellular injury that promotes progression to fibrosis and CKD, and that this model can be used to test mitochondrial therapeutics that limit progression to fibrosis and CKD.
Asunto(s)
Lesión Renal Aguda/inducido químicamente , Lesión Renal Aguda/patología , Ácido Fólico/toxicidad , Hematínicos/toxicidad , Mitocondrias/efectos de los fármacos , Actinas/biosíntesis , Animales , Western Blotting , Colágeno/metabolismo , Creatinina/metabolismo , ADN/biosíntesis , ADN/genética , Complejo IV de Transporte de Electrones/metabolismo , Fibrosis , Glucosuria/inducido químicamente , Homeostasis/efectos de los fármacos , Masculino , Ratones , Mitocondrias/patología , Enfermedades Mitocondriales/inducido químicamente , Enfermedades Mitocondriales/patología , ARN Mensajero/biosíntesis , ARN Mensajero/genéticaRESUMEN
INTRODUCTION: We have examined iron biodistribution and hepatic gene expression in rats following administration of the generic Iron Sucrose Azad (ISA) or the reference iron sucrose drug Venofer®. METHODS: ISA and Venofer® were administered intravenously to normal, non-anemic, male rats at 15 mg/kg (a supra-therapeutic dose-level). To evaluate biodistribution, tissue iron levels were determined over 28 days for plasma, liver, spleen, bone marrow, heart, kidney, lung and stomach using a validated ICP-MS method. Hepatic gene expression was evaluated by microarray analysis of mRNA from samples taken 24 h after drug administration. RESULTS: Iron concentration/time profiles for plasma and tissues were quantitatively similar for ISA and Venofer. Following administration, circulating iron levels briefly exceeded transferrin binding capacity and there was a transient increase in hepatic iron. Bone marrow iron levels remained elevated throughout the study. No increases in tissue iron levels were observed in the heart, stomach or lungs. Spleen iron levels increased over the course of the study in treated and control rats. Small, transient increases were recorded in the kidneys of treated rats. The effects of ISA and Venofer® on hepatic gene transcription were similar. Principal components analysis showed that there was no systematic effect of either treatment on transcriptional profiles. Only a small number of genes showed significant modulation of expression. No transcriptional pattern matches with toxicity pathways were found in the ToxFX database for either treatment. No modulation of key genes in apoptosis, inflammation or oxidative stress pathways was detected. DISCUSSION: These findings demonstrated that the biodistribution of administered iron is essentially similar for Iron Sucrose Azad and Venofer®, that iron sucrose partitions predominantly into the liver, spleen and bone marrow, and that hepatic gene expression studies did not provide any evidence of toxicity in animals treated at a supra-therapeutic dose-level.
Asunto(s)
Medicamentos Genéricos/administración & dosificación , Compuestos Férricos/administración & dosificación , Regulación de la Expresión Génica/efectos de los fármacos , Ácido Glucárico/administración & dosificación , Hematínicos/administración & dosificación , Animales , Relación Dosis-Respuesta a Droga , Medicamentos Genéricos/farmacocinética , Medicamentos Genéricos/toxicidad , Compuestos Férricos/farmacocinética , Compuestos Férricos/toxicidad , Sacarato de Óxido Férrico , Ácido Glucárico/farmacocinética , Ácido Glucárico/toxicidad , Hematínicos/farmacocinética , Hematínicos/toxicidad , Inyecciones Intravenosas , Masculino , Espectrometría de Masas/métodos , Análisis de Componente Principal , Ratas , Ratas Sprague-Dawley , Factores de Tiempo , Distribución TisularRESUMEN
OBJECTIVES: The originator iron polymaltose complex (Maltofer®, IPC, Vifor International, St. Gallen, Switzerland) has been used for over 30 years to treat iron deficiency anemia. Its physico-chemical properties allow for a controlled release of iron, a property which translates into low toxicity and good gastrointestinal (GI) tolerability of the drug compared to the commonly used ferrous salts. A variety of different iron polymaltose complex similars are commercially available with varying structures and, thus, different efficacy and toxicity compared to IPC. In this study, the median lethal dose, the GI tract and liver toxicity of an IPC similar (Vitalix®, IPCSVITA, Laboratorios Roemmers, Buenos Aires, Argentina) were compared with those of IPC in healthy rats. METHODS: The median lethal dose of IPCSVITA was determined as the dose required to kill 6 out of 12 rats after 24 h from dosing. To compare the GI and liver toxicities, rats received IPCSVITA or IPC (both 280 mg iron/kg body weight) for 28 days. GI toxicity was assessed macroscopically by scoring lesion severities and microscopically by analyzing the villi/crypt ratio, number of eosinophils/villi and number of Goblet cells/villi. Ferritin was assessed in the small intestine villi and in the liver by immunostaining. Iron deposits in the liver were assessed by Prussian blue staining. RESULTS: Serum iron concentration and transferrin saturation (TSAT) were significantly higher in the IPCSVITA group vs. the IPC and the control groups. Food consumption, body weight, and bowel movement at Day 29 were significantly lower within the IPCSVITA group vs. the IPC or the control groups. The lesion scores in the stomach and in the lower GI tract of the IPCSVITA group were significantly higher than those of the IPC and control groups. The villi/crypt ratio and the number of Goblet cells/villi in the small intestine were significantly lower in IPCSVITA-treated animals than in IPC-treated or control animals. The number of eosinophils per villi was significantly increased in the IPCSVITA group vs. IPC and control group. In the lower GI tract, microscopic lesions were observed only in the IPCSVITA group. The amount of ferritin in the small intestine and in the liver was higher in IPC-treated animals vs. IPCSVITA- treated or control animals. CONCLUSIONS: Higher serum iron and TSAT levels, lesions in the stomach and lower GI tract suggest the presence of weakly bound iron on the surface of the IPCSVITA complex, which has different physico-chemical properties than IPC. The lower levels of iron deposits in the liver suggest that the iron from IPCSVITA is taken up in a less controlled way than from IPC, thus, potentially accumulating in the wrong cellular compartment.
Asunto(s)
Compuestos Férricos/toxicidad , Tracto Gastrointestinal/efectos de los fármacos , Hematínicos/toxicidad , Hígado/efectos de los fármacos , Animales , Peso Corporal/efectos de los fármacos , Química Farmacéutica , Defecación/efectos de los fármacos , Ingestión de Alimentos/efectos de los fármacos , Femenino , Ferritinas/metabolismo , Tracto Gastrointestinal/metabolismo , Tracto Gastrointestinal/patología , Hierro/sangre , Dosificación Letal Mediana , Hígado/metabolismo , Hígado/patología , Masculino , Ratas , Ratas Sprague-Dawley , Transferrina/metabolismoRESUMEN
OBJECTIVE: Anemia is common in critically ill patients, due to inflammation and blood loss. Anemia can be associated with iron deficiency and low serum hepcidin levels. However, iron administration in this setting remains controversial because of its potential toxicity, including oxidative stress induction and sepsis facilitation. The objective of this work was to determine the efficacy and toxicity of iron administration using a mouse model mimicking critical care anemia as well as a model of acute septicemia. DESIGN: Prospective, randomized, open label controlled animal study. SETTING: University-based research laboratory. SUBJECTS: C57BL/6 and OF1 mice. INTERVENTIONS: Intraperitoneal injection of zymosan inducing generalized inflammation in C57BL/6 mice, followed in our full model by repeated phlebotomies. A dose equivalent to 15 mg/kg of ferric carboxymaltose was injected intravenously on day 5. To assess the toxicity of iron in a septicemia model, OF1 mice were simultaneously injected with iron and different Escherichia coli strains. MEASUREMENTS AND MAIN RESULTS: To investigate the effect of iron on oxidative stress, we measured reactive oxygen species production in the blood using luminol-amplified chemiluminescence and superoxide dismutase 2 messenger RNA levels in the liver. These markers of oxidative stress were increased after iron administration in control mice but not in zymosan-treated mice. Liver catalase messenger RNA levels decreased in iron-treated control mice. Iron administration was not associated with increased mortality in the septicemia model or in the generalized inflammation model. Iron increased hemoglobin levels in mice fed with a low iron diet and subjected to phlebotomies and zymosan 2 wks after treatment administration. CONCLUSIONS: Adverse effects of intravenous iron supplementation by ferric carboxymaltose seem to be minimal in our animal models. Furthermore, iron appears to be effective in correcting anemia, despite inflammation. Studies of efficacy and safety of iron in critically ill patients are warranted.
Asunto(s)
Anemia/tratamiento farmacológico , Compuestos Férricos/administración & dosificación , Compuestos Férricos/toxicidad , Hematínicos/administración & dosificación , Hematínicos/toxicidad , Maltosa/análogos & derivados , Animales , Péptidos Catiónicos Antimicrobianos/genética , Péptidos Catiónicos Antimicrobianos/metabolismo , Catalasa/genética , Catalasa/metabolismo , Dieta , Modelos Animales de Enfermedad , Hemoglobinas , Hepcidinas , Inflamación/inducido químicamente , Inyecciones Intravenosas , Hierro/administración & dosificación , Hierro/análisis , Hígado/química , Hígado/metabolismo , Luminiscencia , Maltosa/administración & dosificación , Maltosa/toxicidad , Ratones , Ratones Endogámicos C57BL , Flebotomía , ARN Mensajero/metabolismo , Distribución Aleatoria , Especies Reactivas de Oxígeno/sangre , Sepsis/tratamiento farmacológico , Bazo/química , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismo , Oligoelementos/administración & dosificación , Zimosan/farmacologíaRESUMEN
Intravenous (i.v.) iron is associated with a risk of oxidative stress. The effects of ferumoxytol, a recently approved i.v. iron preparation, were compared with those of ferric carboxymaltose, low molecular weight iron dextran and iron sucrose in the liver, kidneys and heart of normal rats. In contrast to iron sucrose and ferric carboxymaltose, low molecular weight iron dextran and ferumoxytol caused renal and hepatic damage as demonstrated by proteinuria and increased liver enzyme levels. Higher levels of oxidative stress in these tissues were also indicated, by significantly higher levels of malondialdehyde, significantly increased antioxidant enzyme activities, and a significant reduction in the reduced to oxidized glutathione ratio. Inflammatory markers were also significantly higher with ferumoxytol and low molecular weight iron dextran rats than iron sucrose and ferric carboxymaltose. Polarographic analysis suggested that ferumoxytol contains a component with a more positive reduction potential, which may facilitate iron-catalyzed formation of reactive oxygen species and thus be responsible for the observed effects. Only low molecular weight iron dextran induced oxidative stress and inflammation in the heart.
Asunto(s)
Compuestos Férricos/toxicidad , Hematínicos/toxicidad , Complejo Hierro-Dextran/toxicidad , Nanopartículas de Magnetita/toxicidad , Maltosa/análogos & derivados , Estrés Oxidativo/efectos de los fármacos , Animales , Antioxidantes/análisis , Presión Sanguínea/efectos de los fármacos , Creatinina/metabolismo , Femenino , Compuestos Férricos/administración & dosificación , Sacarato de Óxido Férrico , Óxido Ferrosoférrico , Ácido Glucárico , Corazón/efectos de los fármacos , Hematínicos/administración & dosificación , Inmunohistoquímica , Inflamación/inducido químicamente , Inyecciones Intravenosas , Complejo Hierro-Dextran/administración & dosificación , Riñón/efectos de los fármacos , Riñón/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/enzimología , Nanopartículas de Magnetita/administración & dosificación , Masculino , Maltosa/administración & dosificación , Maltosa/toxicidad , Peso Molecular , Proteinuria/inducido químicamente , Ratas , Ratas Sprague-DawleyRESUMEN
Periodic blood transfusion can lead to secondary iron overload in patients with hematologic and oncologic diseases. Iron overload can result in iron deposition in heart tissue, which decreases cardiac function and can ultimately lead to death due to dilated cardiomyopathy and cardiac failure. In this study, we established murine model of secondary iron overload, studied the changes in cardiac function with echocardiography, and examined the histopathologic changes. Three experimental groups of the six week-old C57/BL mice (H-2(b)) were injected intraperitoneally with 10 mg of iron dextran daily 5 days a week for 2, 4, and 6 weeks. Cumulative doses of iron for the three experimental groups were 100, 200, and 300 mg, while the control groups were injected with the same amounts of phosphate-buffered saline. We studied the cardiac function under anesthesia with echocardiography using a GE Vivid7 Dimension system. Plasma iron levels and liver iron contents were measured. The hearts and livers were harvested and stained with H&E and Perls Prussian blue for iron, and the levels of iron deposit were examined. We assessed the cardiac measurements after adjustment for weight. On echocardiography, thicknesses of the interventricular septum and posterior ventricular wall (PS) during diastole showed correlation with the amount of iron deposit (P < 0.01). End-diastolic volume showed dilatation of the left ventricle in the 300 mg group (P < 0.01). Changes in the fractional shortening were not statistically significant (P = 0.07). Plasma iron levels and liver iron contents were increased proportionally according to the amount of iron loaded. The histopathologic findings of PS and liver showed higher grade of iron deposit proportional to the cumulated iron dose. In this study, we present an animal model which helps understand the cardiac function changes in patients with secondary iron overload due to repeated blood transfusions. Our results may help characterize the pathophysiologic features of cardiomyopathy in patients with secondary iron overload, and our model may be applied to in vivo iron-chelating therapy studies.
Asunto(s)
Cardiomiopatías/fisiopatología , Sobrecarga de Hierro/fisiopatología , Complejo Hierro-Dextran/administración & dosificación , Hierro/metabolismo , Contracción Miocárdica/fisiología , Miocardio/metabolismo , Animales , Cardiomiopatías/etiología , Cardiomiopatías/metabolismo , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Ecocardiografía , Femenino , Hematínicos/administración & dosificación , Hematínicos/toxicidad , Sobrecarga de Hierro/inducido químicamente , Sobrecarga de Hierro/metabolismo , Complejo Hierro-Dextran/toxicidad , Hígado/metabolismo , Hígado/patología , Ratones , Ratones Endogámicos C57BL , Contracción Miocárdica/efectos de los fármacos , Miocardio/patologíaRESUMEN
Intravenous iron preparations are key components in the management of anaemia of various etiologies. These iron-carbohydrate complexes permit safe systemic delivery of iron, whilst protecting from the potential toxic effects of over-saturation. This in turn permits efficient haematopoiesis following erythropoietin administration. Since the rate of release of iron is dependent upon the structure of this iron-carbohydrate complex, it is essential to ensure that an intravenous iron preparation is well characterized and its properties documented. This report describes physicochemical and toxicological studies into a new iron sucrose generic preparation, "Iron Sucrose Azad (ISA)", using the original iron sucrose product as reference. It could be demonstrated that the specifications and physicochemical characteristics of ISA reflect those of the reference product. Furthermore, in a rat model previously shown to identify possible toxicological effects of "unsimilar" iron sucrose preparations, ISA was found to have the same properties as the reference product, with both being well tolerated.
Asunto(s)
Compuestos Férricos/química , Compuestos Férricos/toxicidad , Hematínicos/química , Hematínicos/toxicidad , Algoritmos , Animales , Cromatografía en Gel , Cromatografía Líquida de Alta Presión , Creatinina/metabolismo , Medicamentos Genéricos , Femenino , Compuestos Férricos/farmacocinética , Sacarato de Óxido Férrico , Ácido Glucárico , Hematínicos/farmacocinética , Hierro/sangre , Cinética , Pruebas de Función Hepática , Microscopía de Fuerza Atómica , Peso Molecular , Nefelometría y Turbidimetría , Tamaño de la Partícula , Polarografía , Proteinuria/inducido químicamente , Ratas , Ratas Sprague-Dawley , Estándares de Referencia , Superóxido Dismutasa/metabolismoRESUMEN
Protective effects of adiponectin (APN; an adipocytokine) were shown against various oxidative challenges; however, its therapeutic implications and the mechanisms underlying hepatic iron overload remain unclear. Herein, we show that the deleterious effects of iron dextran on liver function and iron deposition were significantly reversed by adiponectin gene therapy, which was accompanied by AMP-activated protein kinase (AMPK) phosphorylation and heme oxygenase (HO)-1 induction. Furthermore, AMPK-mediated peroxisome proliferator-activated receptor-α (PPARα) activation by APN was ascribable to HO-1 induction. Additionally, we revealed direct transcriptional regulation of HO-1 by the binding of PPARα to a PPAR-responsive element (PPRE) by various experimental assessments. Interestingly, overexpression of HO-1 in hepatocytes mimicked the protective effect of APN in attenuating iron-mediated injury, whereas it was abolished by SnPP and small interfering HO-1. Furthermore, bilirubin, the end-product of the HO-1 reaction, but not CO, protected hepatocytes from iron dextran-mediated caspase activation. Herein, we demonstrate a novel functional PPRE in the promoter regions of HO-1, and APN-mediated HO-1 induction elicited an antiapoptotic effect and a decrease in iron deposition in hepatocytes subjected to iron challenge.