RESUMEN
Experimental and theoretical study of the influence of the pump-to-laser beam overlap, a crucial parameter for optimization of optically pumped alkali atom lasers, is reported for Ti:Sapphire pumped Cs laser. Maximum laser power > 370 mW with an optical-to-optical efficiency of 43% and slope efficiency ~55% was obtained. The dependence of the lasing power on the pump power was found for different pump beam radii at constant laser beam radius. Non monotonic dependence of the laser power (optimized over the temperature of the Cs cell) on the pump beam radius was observed with a maximum achieved at the ratio ~0.7 between the pump and laser beam radii. The optimal temperature decreased with increasing pump beam radius. A simple optical model of the laser, where Gaussian spatial shapes of the pump and laser intensities in any cross section of the beams were assumed, was compared to the experiments. Good agreement was obtained between the measured and calculated dependence of the laser power on the pump power at different pump beam radii and also of the laser power, threshold pump power and optimal temperature on the pump beam radius. The model does not use empirical parameters such as mode overlap efficiency and can be applied to different Ti:Sapphire and diode pumped alkali lasers with arbitrary spatial distributions of the pump and laser beam widths.
RESUMEN
The principal finding of this study is that two drugs, alverine and benfluorex, used in vastly different clinical settings, activated the nuclear receptor transcription factor HNF4α. Both were hits in a high-throughput screen for compounds that reversed the inhibitory effect of the fatty acid palmitate on human insulin promoter activity. Alverine is used in the treatment of irritable bowel syndrome, while benfluorex (Mediator) was used to treat hyperlipidemia and type II diabetes. Benfluorex was withdrawn from the market recently because of serious cardiovascular side effects related to fenfluramine-like activity. Strikingly, alverine and benfluorex have a previously unrecognized structural similarity, consistent with a common mechanism of action. Gene expression and biochemical studies revealed that they both activate HNF4α. This novel mechanism of action should lead to a reinterpretation of previous studies with these drugs and suggests a path toward the development of therapies for diseases such as inflammatory bowel and diabetes that may respond to HNF4α activators.
Asunto(s)
Fenfluramina/análogos & derivados , Factor Nuclear 4 del Hepatocito/metabolismo , Propilaminas/química , Línea Celular , Relación Dosis-Respuesta a Droga , Evaluación Preclínica de Medicamentos , Fenfluramina/química , Fenfluramina/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Modelos Moleculares , Estructura Molecular , Propilaminas/farmacología , Unión Proteica/efectos de los fármacosRESUMEN
Hepatocyte nuclear factor (HNF)4α is a central regulator of gene expression in cell types that play a critical role in metabolic homeostasis, including hepatocytes, enterocytes, and pancreatic ß cells. Although fatty acids were found to occupy the HNF4α ligand-binding pocket and were proposed to act as ligands, there is controversy about both the nature of HNF4α ligands as well as the physiological role of the binding. Here, we report the discovery of potent synthetic HNF4α antagonists through a high-throughput screen for effectors of the human insulin promoter. These molecules bound to HNF4α with high affinity and modulated the expression of known HNF4α target genes. Notably, they were found to be selectively cytotoxic to cancer cell lines in vitro and in vivo, although in vivo potency was limited by suboptimal pharmacokinetic properties. The discovery of bioactive modulators for HNF4α raises the possibility that diseases involving HNF4α, such as diabetes and cancer, might be amenable to pharmacologic intervention by modulation of HNF4α activity.
Asunto(s)
Bencimidazoles/farmacología , Descubrimiento de Drogas , Factor Nuclear 4 del Hepatocito/antagonistas & inhibidores , Ensayos Analíticos de Alto Rendimiento , Insulina/genética , Regiones Promotoras Genéticas/genética , Sulfonamidas/farmacología , Bencimidazoles/química , Relación Dosis-Respuesta a Droga , Factor Nuclear 4 del Hepatocito/genética , Factor Nuclear 4 del Hepatocito/metabolismo , Humanos , Modelos Moleculares , Estructura Molecular , PPAR gamma/agonistas , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Relación Estructura-Actividad , Sulfonamidas/químicaRESUMEN
A number of diabetogenic stimuli interact to influence insulin promoter activity, making it an attractive target for both mechanistic studies and therapeutic interventions. High-throughput screening (HTS) for insulin promoter modulators has the potential to reveal novel inputs into the control of that central element of the pancreatic beta-cell. A cell line from human islets in which the expression of insulin and other beta-cell-restricted genes are modulated by an inducible form of the bHLH transcription factor E47 was developed. This cell line, T6PNE, was adapted for HTS by transduction with a vector expressing green fluorescent protein under the control of the human insulin promoter. The resulting cell line was screened against a library of known drugs for those that increase insulin promoter activity. Members of the phenothiazine class of neuroleptics increased insulin gene expression upon short-term exposure. Chronic treatment, however, resulted in suppression of insulin promoter activity, consistent with the effect of phenothiazines observed clinically to induce diabetes in chronically treated patients. In addition to providing insights into previously unrecognized targets and mechanisms of action of phenothiazines, the novel cell line described here provides a broadly applicable platform for mining new molecular drug targets and central regulators of beta-cell differentiated function.