RESUMEN
OBJECTIVES: To evaluate the efficacy and safety of different doses and regimens of filgotinib, an oral Janus kinase 1 inhibitor, as add-on treatment to methotrexate (MTX) in patients with active rheumatoid arthritis (RA) and inadequate response to MTX. METHODS: In this 24-week phase IIb study, patients with moderate-to-severe active RA receiving a stable dose of MTX were randomised (1:1:1:1:1:1:1) to receive placebo or 50, 100 or 200â mg filgotinib, administered once daily or twice daily. Primary end point was the percentage of patients achieving a week 12 American College of Rheumatology (ACR)20 response. RESULTS: Overall, 594 patients were randomised and treated. At week 12, significantly more patients receiving filgotinib 100â mg once daily or 200â mg daily (both regimens) achieved an ACR20 response versus placebo. For other key end points at week 12 (ACR50, ACR-N, Disease Activity Score based on 28 joints and C reactive protein value, Clinical Disease Activity Index, Simplified Disease Activity Index and Health Assessment Questionnaire-Disability Index), differences in favour of 100â or 200â mg filgotinib daily were seen versus placebo; responses were maintained or improved through to week 24. Rapid onset of action and dose-dependent responses were observed for most efficacy end points and were associated with an increased haemoglobin concentration. No significant differences between once-daily and twice-daily regimens were seen. Treatment-emergent adverse event rates were similar in placebo and filgotinib groups. Serious infections occurred in one and five patients in the placebo and filgotinib groups, respectively. No tuberculosis or opportunistic infections were reported. CONCLUSIONS: Filgotinib as add-on to MTX improved the signs and symptoms of active RA over 24â weeks and was associated with a rapid onset of action. Filgotinib was generally well tolerated. TRIAL REGISTRATION NUMBER: NCT01888874.
Asunto(s)
Artritis Reumatoide/tratamiento farmacológico , Inhibidores de Proteínas Quinasas/administración & dosificación , Piridinas/administración & dosificación , Triazoles/administración & dosificación , Administración Oral , Antirreumáticos/uso terapéutico , Artritis Reumatoide/sangre , Proteína C-Reactiva/metabolismo , Evaluación de la Discapacidad , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Quimioterapia Combinada/efectos adversos , Femenino , Hemoglobinas/metabolismo , Humanos , Infecciones/inducido químicamente , Janus Quinasa 1/antagonistas & inhibidores , Masculino , Metotrexato/uso terapéutico , Persona de Mediana Edad , Inhibidores de Proteínas Quinasas/efectos adversos , Piridinas/efectos adversos , Índice de Severidad de la Enfermedad , Encuestas y Cuestionarios , Triazoles/efectos adversosRESUMEN
OBJECTIVES: To evaluate the efficacy and safety of different doses of filgotinib, an oral Janus kinase 1 inhibitor, as monotherapy in patients with active rheumatoid arthritis (RA) and previous inadequate response to methotrexate (MTX). METHODS: In this 24-week phase IIb study, patients with moderately to severely active RA were randomised (1:1:1:1) to receive 50, 100 or 200â mg filgotinib once daily, or placebo, after a ≥4-week washout from MTX. The primary end point was the percentage of patients achieving an American College of Rheumatology (ACR)20 response at week 12. RESULTS: Overall, 283 patients were randomised and treated. At week 12, significantly more patients receiving filgotinib at any dose achieved ACR20 responses versus placebo (≥65% vs 29%, p<0.001). For other key end points at week 12 (ACR50, ACR70, ACR-N, Disease Activity Score based on 28 joints and C reactive protein, Clinical Disease Activity Index, Simplified Disease Activity Index and Health Assessment Questionnaire-Disability Index) significant differences from baseline in favour of filgotinib 100 and 200â mg versus placebo were seen; responses were maintained or improved through week 24. Rapid onset of action was observed for most efficacy end points. Dose-dependent increases in haemoglobin were observed. The percentage of patients with treatment-emergent adverse events (TEAE) was similar in the placebo and filgotinib groups (â¼40%). Eight patients on filgotinib and one on placebo had a serious TEAE, and four patients, all of whom received filgotinib, experienced a serious infection. No tuberculosis or opportunistic infections were reported. CONCLUSIONS: Over 24â weeks, filgotinib as monotherapy was efficacious in treating the signs and symptoms of active RA, with a rapid onset of action. Filgotinib was generally well tolerated. TRIAL REGISTRATION NUMBER: NCT01894516.
Asunto(s)
Artritis Reumatoide/tratamiento farmacológico , Inhibidores de Proteínas Quinasas/administración & dosificación , Piridinas/administración & dosificación , Triazoles/administración & dosificación , Administración Oral , Antirreumáticos/uso terapéutico , Artritis Reumatoide/sangre , Proteína C-Reactiva/metabolismo , Evaluación de la Discapacidad , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Femenino , Hemoglobinas/metabolismo , Humanos , Infecciones/inducido químicamente , Janus Quinasa 1/antagonistas & inhibidores , Masculino , Metotrexato/uso terapéutico , Persona de Mediana Edad , Inhibidores de Proteínas Quinasas/efectos adversos , Piridinas/efectos adversos , Retratamiento , Índice de Severidad de la Enfermedad , Encuestas y Cuestionarios , Resultado del Tratamiento , Triazoles/efectos adversosRESUMEN
Developing new sources of organic Se has potential benefit for animal production and human nutrition via animal-based foods enriched in Se. The objectives of this trial were to compare L-selenomethionine with another organic Se source, Se-enriched yeast (SelPlex 2300), and sodium selenite, an inorganic Se source, against a commercial control diet. The effect of source and the dosage of Se supplementation on Se in eggs and blood variables was investigated. Ten treatments were used with 18 laying hens per group. In addition to the control diet, the control diet was supplemented with L-selenomethionine, Se-enriched yeast, or sodium selenite at 0.1, 0.3, or 0.5 mg/kg of Se. The feeding trial lasted 8 wk. Birds in the different treatment groups all showed good performance. At d 0 and 56, Se and glutathione peroxidase (GPx) were analyzed in 10 blood samples per group. After supplementing the diets for 56 d, significantly higher Se levels in serum and egg contents were reached for the Se-supplemented groups compared with the control. Supplementing 0.3 and 0.5 mg/kg of L-selenomethionine or Se-enriched yeast instead of 0.1 mg/kg significantly increased the serum Se levels, whereas no significant increase was found for sodium selenite. No effect of Se source or dosage was observed on serum GPx levels. Selenium in eggs was significantly affected by dosage and source of Se. The Se supplementation level in the feed was reflected in the eggs, with the highest and lowest values for 0.5 and 0.1 mg/kg, respectively, and values in between for the 0.3 mg/kg supplementation level. A dose response was most pronounced for L-selenomethionine, followed by Se-enriched yeast, and was least when Se was added as sodium selenite. It can be concluded that Se from organic sources was more bioavailable than the inorganic Se source as evidenced by blood and egg Se levels. Within the organic Se sources, L-selenomethionine showed higher Se transfer to eggs than Se-enriched yeast.
Asunto(s)
Pollos/fisiología , Oviposición/efectos de los fármacos , Selenometionina/farmacología , Selenito de Sodio/farmacología , Levaduras , Alimentación Animal/análisis , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Dieta/veterinaria , Suplementos Dietéticos , Femenino , Oviposición/fisiología , Selenometionina/administración & dosificación , Selenito de Sodio/administración & dosificaciónRESUMEN
In many pharmacological and toxicological studies knowledge about the intestinal absorption, which is dependent upon the surface area of absorptive epithelia, is indispensible. Although mice are often used in such preclinical studies, very few quantitative data about their intestinal surface area are available. Especially for locally acting candidate drugs in development, this information is crucial for dose translation towards humans. Therefore, the surface area of the intestinal tract of CD-1 IGS mice was assessed in the present study. The intestinal tracts of 12 mice were collected after euthanasia. From six animals, histological sections from the duodenum, jejunum, ileum, caecum and colon-rectum were made according to common stereological principles. Using these sections, the volumes and surface areas of each intestinal segment were estimated applying stereological counting procedures. In the other six animals, the density and surface area of the microvilli present in each intestinal segment were determined by means of scanning and transmission electron microscopy to assess the increase of the intestinal surface area attributable to the presence of microvilli. The mean total volume and surface area of the intestinal tract were 1.34 cm(3) and 1.41 m(2), respectively. The relative intestinal surface area (intestinal surface area divided by the body surface area) was 119. The relative intestinal surface area of mice is very similar to that of humans. The results of this study are important for the appropriate dose translation of candidate therapeutic compounds in drug development from mouse to humans.
Asunto(s)
Enterocitos/ultraestructura , Absorción Intestinal/fisiología , Intestinos/anatomía & histología , Xenobióticos/farmacocinética , Administración Oral , Animales , Relación Dosis-Respuesta a Droga , Enterocitos/metabolismo , Femenino , Humanos , Mucosa Intestinal/metabolismo , Masculino , Ratones , Ratones Endogámicos , Microscopía Electrónica de Rastreo , Microscopía Electrónica de Transmisión , Microvellosidades/metabolismo , Microvellosidades/ultraestructura , Tamaño de los Órganos , Especificidad de la EspecieRESUMEN
Myelin-reactive T cells are considered to play an essential role in the pathogenesis of multiple sclerosis (MS), an autoimmune disease of the central nervous system. We have previously studied the effects of T cell vaccination (TCV), a procedure by which MS patients are immunized with attenuated autologous myelin basic protein (MBP)-reactive T cell clones. Because several myelin antigens are described as potential autoantigens for MS, T cell vaccines incorporating a broad panel of antimyelin reactivities may have therapeutic effects. Previous reports have shown an accumulation of activated T cells recognizing multiple myelin antigens in the cerebrospinal fluid (CSF) of MS patients. We conducted a pilot clinical trial of TCV with activated CD4+ T cells derived from CSF in five MS patients (four RR, one CP) to study safety, feasibility and immune effects of TCV. CSF lymphocytes were cultured in the presence of rIL-2 and depleted for CD8 cells. After 5-8 weeks CSF T cell lines (TCL) were almost pure TCR alpha beta+CD4+ cells of the Th1/Th0 type. The TCL showed reactivity to MBP, MOG and/or PLP as tested by Elispot and had a restricted clonality. Three immunizations with irradiated CSF vaccines (10 million cells) were administered with an interval of 2 months. The vaccinations were tolerated well and no toxicity or adverse effects were reported. The data from this small open-label study cannot be used to support efficacy. However, all patients remained clinically stable or had reduced EDSS with no relapses during or after the treatment. Proliferative responses against the CSF vaccine were observed in 3/5 patients. Anti-ergotypic responses were observed in all patients. Anti-MBP/PLP/MOG reactivities remained low or were reduced in all patients. Based on these encouraging results, we recently initiated a double-blind placebo-controlled trial with 60 MS patients to study the effects of TCV with CSF-derived vaccines in early RR MS patients.
Asunto(s)
Traslado Adoptivo/métodos , Linfocitos T CD4-Positivos/inmunología , Activación de Linfocitos , Esclerosis Múltiple/terapia , Proteína Básica de Mielina/inmunología , Receptores de Antígenos de Linfocitos T alfa-beta/inmunología , Adulto , Autoantígenos/inmunología , División Celular , Células Clonales , Femenino , Genes Codificadores de la Cadena beta de los Receptores de Linfocito T , Humanos , Recuento de Linfocitos , Masculino , Persona de Mediana Edad , Esclerosis Múltiple/líquido cefalorraquídeo , Esclerosis Múltiple/inmunología , Proyectos Piloto , Complejo Receptor-CD3 del Antígeno de Linfocito T/inmunologíaRESUMEN
In this study, analogues of olomoucine, a previously described plant cytokinin analogue with cyclin-dependent kinase (CDK) inhibitory activity, were investigated for effect on CDK1 and CDK2 and for effect on cell proliferation. Eight new compounds exhibit stronger inhibitory activity on CDK1 and CDK2 and on cell proliferation than olomoucine. Some active compounds showed low inhibition of proliferation of normal myeloid growth. Improvement of inhibitory activity of known compounds with a C6-benzylamino group was brought about by substitution with one hydroxyl. Also, new C2 substituents associated with inhibitory activity on CDK and on cell proliferation are described. There was a significant correlation between effect on CDK and antiproliferative effect on the KG1 and Molt3 cell lines and on primary human lymphocytes, strongly suggesting that at least part of the antiproliferative effect of cytokinin analogues was due to inhibition of CDK activity. Cytokinin analogues induced apoptosis in a time- and concentration-dependent manner and changes in cell cycle distribution. The antiproliferative and pro-apoptotic effects of plant cytokinin analogues suggest that they are a new class of cytostatic agents and that they may find an application in the chemotherapy of cancer.
Asunto(s)
Proteína Quinasa CDC2/antagonistas & inhibidores , Quinasas CDC2-CDC28 , Quinasas Ciclina-Dependientes/antagonistas & inhibidores , Citocininas/farmacología , Inhibidores Enzimáticos/farmacología , Leucemia/patología , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Purinas/farmacología , Apoptosis/efectos de los fármacos , Proteína Quinasa CDC2/metabolismo , Ciclo Celular/efectos de los fármacos , División Celular/efectos de los fármacos , Ensayo de Unidades Formadoras de Colonias , Quinasa 2 Dependiente de la Ciclina , Quinasas Ciclina-Dependientes/metabolismo , ADN de Neoplasias/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Cinetina , Leucemia/enzimología , Plantas , Proteínas Serina-Treonina Quinasas/metabolismo , Factores de Tiempo , Células Tumorales Cultivadas/efectos de los fármacos , Células Tumorales Cultivadas/enzimologíaRESUMEN
The analysis of T cell receptor variable (TCR V) gene repertoires in blood or tissues may provide important information when studying immunopathological mechanisms. The overexpression of a TCR gene may indicate the expansion of the corresponding T cell subset. In autoimmune diseases, clonally expanded T cell subsets in the affected organs may represent pathogenic lymphocytes. We describe a simple, rapid and sensitive method to determine the TCR AV and BV gene repertoire using a PCR-ELISA method. RNA is extracted from lymphocytes, transcribed to cDNA, which is then used as a template for PCR with 19 different TCR AV gene and 20 BV gene specific primers as the forward primer, and a digoxigenin (DIG) labeled AC/BC primer as the reverse primer. The DIG labeled PCR amplicons are hybridized with a fluorescein isothiocyanate (FITC) labeled TCR C region specific probe. Finally, the amplicons are quantified by ELISA using anti-FITC coated microtiter plates, and an anti-DIG conjugated peroxidase. Although PCR-ELISA cannot accurately quantify the expression level of a given TCR gene, overrepresented TCR V genes are easily identified by comparing the relative expression levels of each individual V gene in the total V gene repertoire. We demonstrate that this technique can be used to determine TCR profiles in blood and tissue samples containing as few as 50,000 T cells. In combination with CDR3 fragment size analysis, this method is an efficient tool to identify clonally expanded T cell subsets in the synovial biopsies of rheumatoid arthritis patients.
Asunto(s)
Artritis Reumatoide/patología , Toxinas Bacterianas , Reordenamiento Génico de Linfocito T , Genes Codificadores de la Cadena beta de los Receptores de Linfocito T , Reacción en Cadena de la Polimerasa/métodos , Receptores de Antígenos de Linfocitos T/sangre , Receptores de Antígenos de Linfocitos T/genética , Superantígenos , Artritis Reumatoide/sangre , Artritis Reumatoide/inmunología , Células Clonales , Cartilla de ADN , Enterotoxinas/farmacología , Ensayo de Inmunoadsorción Enzimática , Humanos , Células Jurkat , Cinética , Activación de Linfocitos , Recuento de Linfocitos , Fitohemaglutininas/farmacología , Reproducibilidad de los Resultados , Staphylococcus aureus/inmunología , Líquido Sinovial/inmunología , Membrana Sinovial/inmunología , Membrana Sinovial/patología , Subgrupos de Linfocitos T/metabolismo , Subgrupos de Linfocitos T/patologíaRESUMEN
This study was undertaken to examine the contribution of uncontained morsellized bone graft to the structural properties of a femoral reconstruction in total knee arthroplasty and to serve as a basis for an in vivo animal study. Ten human distal femora with a standard unicondylar uncontained medial bone defect were prepared to fit a femoral component of a cruciate sacrificing TKA. A cyclic axial load of 750 N was applied to the medial part of the femoral component in the presence of impacted morsellized bone graft. After removal of the bone graft, the cyclic loading was repeated for the unsupported situation. None of the grafts collapsed and all cement mantles stayed intact during the experiments. Elastic deformation during cyclic loading was significantly less when graft was added while time-dependent deformation was not affected. We conclude that impacted morsellized bone graft, used for reconstruction of uncontained femoral bone loss in revision knee arthroplasty, may improve the structural resistance against loading. Further animal experimentation for in vivo application is warranted.