RESUMEN

Previous data have shown that RXR-selective agonists (e.g., 3 and 4) are insulin sensitizers in rodent models of non-insulin-dependent diabetes mellitus (NIDDM). Unfortunately, they also produce dramatic increases in triglycerides and profound suppression of the thyroid hormone axis. Here we describe the design and synthesis of new RXR modulators that retain the insulin-sensitizing activity of RXR agonists but produce substantially reduced side effects. These molecules bind selectively and with high affinity to RXR and, unlike RXR agonists, do not activate RXR homodimers. To further evaluate the antidiabetic activity of these RXR modulators, we have designed a concise and systematic structure-activity relationship around the 2E,4E,6Z-7-aryl-3-methylocta-2,4,6-trienoic acid scaffold. Selected compounds have been evaluated using insulin-resistant rodents (db/db mice) to characterize effects on glucose homeostasis. Our studies demonstrate the effectiveness of RXR modulators in lowering plasma glucose in the db/db mouse model.

Asunto(s)

Caprilatos/síntesis química , Diabetes Mellitus Tipo 2/sangre , Hipoglucemiantes/síntesis química , Receptores de Ácido Retinoico/efectos de los fármacos , Factores de Transcripción/efectos de los fármacos , Animales , Glucemia/análisis , Caprilatos/química , Caprilatos/farmacología , Hipoglucemiantes/química , Hipoglucemiantes/farmacología , Resistencia a la Insulina , Masculino , Ratones , Ensayo de Unión Radioligante , Ratas , Ratas Sprague-Dawley , Receptores de Ácido Retinoico/metabolismo , Receptores X Retinoide , Relación Estructura-Actividad , Factores de Transcripción/metabolismo

RESUMEN

The RXR serves as a heterodimer partner for the PPARgamma and the dimer is a molecular target for insulin sensitizers such as the thiazolidinediones. Ligands for either receptor can activate PPAR-dependent pathways via PPAR response elements. Unlike PPARgamma agonists, however, RXR agonists like LG100268 are promiscuous and activate multiple RXR heterodimers. Here, we demonstrate that LG100754, a RXR:RXR antagonist and RXR:PPARalpha agonist, also functions as a RXR:PPARgamma agonist. It does not activate other LG100268 responsive heterodimers like RXR:liver X receptoralpha, RXR:liver X receptorbeta, RXR:bile acid receptor/farnesoid X receptor and RXR:nerve growth factor induced gene B. This unique RXR ligand triggers cellular RXR:PPARgamma-dependent pathways including adipocyte differentiation and inhibition of TNFalpha-mediated hypophosphorylation of the insulin receptor, but does not activate key farnesoid X receptor and liver X receptor target genes. Also, LG100754 treatment of db/db animals leads to an improvement in insulin resistance in vivo. Interestingly, activation of RXR:PPARgamma by LG100268 and LG100754 occurs through different mechanisms. Therefore, LG100754 represents a novel class of insulin sensitizers that functions through RXR but exhibits greater heterodimer selectivity compared with LG100268. These results establish an approach to the design of novel RXR-based insulin sensitizers with greater specificity.

Asunto(s)

Glucemia/metabolismo , Receptores Citoplasmáticos y Nucleares/agonistas , Receptores de Ácido Retinoico/agonistas , Retinoides/farmacología , Tetrahidronaftalenos/farmacología , Factores de Transcripción/agonistas , Células 3T3 , Adipocitos/efectos de los fármacos , Adipocitos/metabolismo , Animales , Diferenciación Celular/efectos de los fármacos , Diabetes Mellitus/sangre , Diabetes Mellitus/tratamiento farmacológico , Dimerización , Resistencia a la Insulina , Ratones , Ácidos Nicotínicos/farmacología , Fosforilación , Receptor de Insulina/metabolismo , Receptores Citoplasmáticos y Nucleares/efectos de los fármacos , Receptores Citoplasmáticos y Nucleares/fisiología , Receptores de Ácido Retinoico/efectos de los fármacos , Receptores de Ácido Retinoico/fisiología , Receptores X Retinoide , Retinoides/uso terapéutico , Tetrahidronaftalenos/uso terapéutico , Factores de Transcripción/efectos de los fármacos , Factores de Transcripción/fisiología , Factor de Necrosis Tumoral alfa/farmacología

RESUMEN

Both retinoid X receptor (RXR)-selective agonists (rexinoids) and thiazolidinediones (TZDs), PPAR (peroxisome proliferator-activated receptor)-gamma-specific ligands, produce insulin sensitization in diabetic rodents. In vitro studies have demonstrated that TZDs mediate their effects via the RXR/PPAR-gamma complex. To determine whether rexinoids lower hyperglycemia by activating the RXR/PPAR-gamma heterodimer in vivo, we compared the effects of a rexinoid (LG100268) and a TZD (rosiglitazone) on gene expression in white adipose tissue, skeletal muscle, and liver of Zucker diabetic fatty rats (ZDFs). In adipose tissue, rosiglitazone decreased tumor necrosis factor-alpha (TNF-alpha) mRNA and induced glucose transporter 4 (GLUT4), muscle carnitine palmitoyl-transferase (MCPT), stearoyl CoA desaturase (SCD1), and fatty acid translocase (CD36). In contrast, LG100268 increased TNF-alpha and had no effect or suppressed the expression of GLUT4, MCPT, SCD1, and CD36. In liver, the rexinoid increased MCPT, SCD1, and CD36 mRNAs, whereas rosiglitazone induced only a small increase in CD36. In skeletal muscle, rosiglitazone and LG100268 have similar effects; both increased SCD1 and CD36 mRNAs. The differences in the pattern of genes induced by the rexinoids and the TZDs in diabetic animals found in these studies suggests that these compounds may have independent and tissue-specific effects on metabolic control in vivo.

Asunto(s)

Diabetes Mellitus Experimental/metabolismo , Hipoglucemiantes/farmacología , Ácidos Nicotínicos/farmacología , Tetrahidronaftalenos/farmacología , Tiazoles/farmacología , Tiazolidinedionas , Tejido Adiposo/efectos de los fármacos , Tejido Adiposo/metabolismo , Animales , Glucemia/efectos de los fármacos , Diabetes Mellitus/tratamiento farmacológico , Diabetes Mellitus/metabolismo , Diabetes Mellitus Experimental/complicaciones , Diabetes Mellitus Experimental/tratamiento farmacológico , Modelos Animales de Enfermedad , Expresión Génica/efectos de los fármacos , Prueba de Tolerancia a la Glucosa , Hiperglucemia/sangre , Hiperglucemia/tratamiento farmacológico , Hiperglucemia/etiología , Hiperinsulinismo/sangre , Hiperinsulinismo/tratamiento farmacológico , Hiperinsulinismo/etiología , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/metabolismo , Obesidad , ARN Mensajero/análisis , Ratas , Ratas Zucker , Receptores Citoplasmáticos y Nucleares/metabolismo , Receptores de Ácido Retinoico/agonistas , Receptores de Ácido Retinoico/genética , Receptores de Ácido Retinoico/metabolismo , Receptores X Retinoide , Rosiglitazona , Factores de Transcripción/agonistas , Factores de Transcripción/genética , Factores de Transcripción/metabolismo

RESUMEN

Hypertriglyceridemia is a frequent complication accompanying the treatment of patients with either retinoids or rexinoids, [retinoid X receptor (RXR)-selective retinoids]. To investigate the cellular and molecular basis for this observation, we have studied the effects of rexinoids on triglyceride metabolism in both normal and diabetic rodents. Administration of a rexinoid such as LG100268 (LG268) to normal or diabetic rats results in a rapid increase in serum triglyceride levels. LG268 has no effect on hepatic triglyceride production but suppresses post-heparin plasma lipoprotein lipase (LPL) activity suggesting that the hypertriglyceridemia results from diminished peripheral processing of plasma very low density lipoproteins particles. Treatment of diabetic rats with rexinoids suppresses skeletal and cardiac muscle but not adipose tissue LPL activity. This effect is independent of changes in LPL mRNA. In C2C12 myocytes, LG268 suppresses the level of cell surface (i.e., heparin-releasable) LPL activity without altering LPL mRNA. This effect is very rapid (t(1/2) = 2 h) and is blocked by the transcriptional inhibitor actinomycin D. These studies demonstrate that RXR ligands can have dramatic effects on the post-translational processing of LPL and suggest that skeletal muscle may be an important target of rexinoid action. In addition, these data underscore that the metabolic consequences of RXR activation are distinct from either retinoic acid receptor or peroxisome proliferator-activated receptor activation.

Asunto(s)

Lipoproteína Lipasa/metabolismo , Ácidos Nicotínicos/farmacología , Receptores de Ácido Retinoico/metabolismo , Tetrahidronaftalenos/farmacología , Factores de Transcripción/metabolismo , Animales , Células Cultivadas , Corazón/efectos de los fármacos , Hipertrigliceridemia/sangre , Hipertrigliceridemia/inducido químicamente , Lipoproteína Lipasa/efectos de los fármacos , Lipoproteínas VLDL/efectos de los fármacos , Lipoproteínas VLDL/metabolismo , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/enzimología , Miocardio/enzimología , Miocardio/metabolismo , Ácidos Nicotínicos/efectos adversos , Ratas , Ratas Sprague-Dawley , Ratas Zucker , Receptores de Ácido Retinoico/efectos de los fármacos , Receptores X Retinoide , Retinoides , Tetrahidronaftalenos/efectos adversos , Factores de Transcripción/efectos de los fármacos , Triglicéridos/sangre

RESUMEN

A common feature of many metabolic pathways is their control by retinoid X receptor (RXR) heterodimers. Dysregulation of such metabolic pathways can lead to the development of atherosclerosis, a disease influenced by both systemic and local factors. Here we analyzed the effects of activation of RXR and some of its heterodimers in apolipoprotein E -/- mice, a well established animal model of atherosclerosis. An RXR agonist drastically reduced the development of atherosclerosis. In addition, a ligand for the peroxisome proliferator-activated receptor (PPAR)gamma and a dual agonist of both PPARalpha and PPARgamma had moderate inhibitory effects. Both RXR and liver X receptor (LXR) agonists induced ATP-binding cassette protein 1 (ABC-1) expression and stimulated ABC-1-mediated cholesterol efflux from macrophages from wild-type, but not from LXRalpha and beta double -/-, mice. Hence, activation of ABC-1-mediated cholesterol efflux by the RXR/LXR heterodimer might contribute to the beneficial effects of rexinoids on atherosclerosis and warrant further evaluation of RXR/LXR agonists in prevention and treatment of atherosclerosis.

Asunto(s)

Apolipoproteínas E/fisiología , Arteriosclerosis/prevención & control , Receptores de Ácido Retinoico/metabolismo , Factores de Transcripción/metabolismo , Transportadoras de Casetes de Unión a ATP/fisiología , Animales , Apolipoproteínas E/genética , Arteriosclerosis/genética , Transporte Biológico , Colesterol/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Receptores X Retinoide

RESUMEN

Several nuclear hormone receptors involved in lipid metabolism form obligate heterodimers with retinoid X receptors (RXRs) and are activated by RXR agonists such as rexinoids. Animals treated with rexinoids exhibited marked changes in cholesterol balance, including inhibition of cholesterol absorption and repressed bile acid synthesis. Studies with receptor-selective agonists revealed that oxysterol receptors (LXRs) and the bile acid receptor (FXR) are the RXR heterodimeric partners that mediate these effects by regulating expression of the reverse cholesterol transporter, ABC1, and the rate-limiting enzyme of bile acid synthesis, CYP7A1, respectively. Thus, these RXR heterodimers serve as key regulators of cholesterol homeostasis by governing reverse cholesterol transport from peripheral tissues, bile acid synthesis in liver, and cholesterol absorption in intestine.

Asunto(s)

Transportadoras de Casetes de Unión a ATP/metabolismo , Colesterol/metabolismo , Glicoproteínas/metabolismo , Absorción Intestinal/efectos de los fármacos , Intestino Delgado/metabolismo , Hígado/metabolismo , Receptores Citoplasmáticos y Nucleares , Receptores de Ácido Retinoico/metabolismo , Factores de Transcripción/metabolismo , Transportador 1 de Casete de Unión a ATP , Transportadoras de Casetes de Unión a ATP/genética , Animales , Ácidos y Sales Biliares/biosíntesis , Transporte Biológico/efectos de los fármacos , Colesterol 7-alfa-Hidroxilasa/metabolismo , Colesterol en la Dieta/administración & dosificación , Cricetinae , Proteínas de Unión al ADN/metabolismo , Dimerización , Regulación de la Expresión Génica/efectos de los fármacos , Glicoproteínas/genética , Homeostasis/efectos de los fármacos , Ligandos , Receptores X del Hígado , Macrófagos Peritoneales/metabolismo , Masculino , Mesocricetus , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Receptores Nucleares Huérfanos , Receptores de Ácido Retinoico/agonistas , Receptores de Ácido Retinoico/genética , Receptores de Hormona Tiroidea/agonistas , Receptores de Hormona Tiroidea/genética , Receptores de Hormona Tiroidea/metabolismo , Receptores X Retinoide , Factores de Transcripción/agonistas

RESUMEN

OBJECTIVE: To investigate whether retinoid X receptor agonists act as insulin sensitizers and compare their effects with that of thiazolidinedione BRL 49653 in obese Zucker rats. DESIGN: In two independent studies, obese Zucker rats were dosed orally once daily for 14 days with one of the following treatments: LG 100268 (20 mg/kg), LG 100324 (20 mg/kg), BRL 49653 (3 mg/kg) or vehicle. MEASUREMENTS: Daily food intake and body weight gain, blood glucose, plasma and pancreatic insulin, whole body glucose disposal (by euglycaemic-hyperinsulinaemic clamp) and tissue glucose utilization. RESULTS: The retinoid X receptor agonists (rexinoids) LG 100268 and LG 100324 caused a reduction in the food intake of obese Zucker rats relative to controls and to rats receiving BRL 49653. The two rexinoids also produced a marked decrease in the body weight gain, whereas the growth rate of rats treated with BRL 49653 tended to increase. Both rexinoids and BRL 49653 reduced the plasma insulin concentration of fed rats. LG 100268 and LG 100324 also significantly lowered blood glucose concentrations after 1 week of treatment. The 5 h fasted plasma insulin concentration was significantly lower in the rexinoid-treated groups and the terminal insulin level (at the end of the clamp) tended to be lower in all treated groups compared with animals given the dosing vehicle. However, pancreatic insulin content was not affected by any of the treatments. Under euglycaemic-hyperinsulinaemic clamp conditions, there were no significant differences in the rate of hepatic glucose output and whole body glucose disposal, except that, in experiment 1, BRL 49653 caused significant increase in the glucose infusion rate and muscle glucose utilization. In experiment 2, a similar glucose infusion rate to the controls was achieved in all treatment groups but the steady-state insulin concentration in the treated animals was only about 50% of that in the control animals, despite the fact that all rats received a similar insulin infusion concentration. This suggests that both the rexinoids and BRL 49653 increased insulin clearance. CONCLUSIONS: Chronic administration of retinoid X receptor agonists LG 100268 and LG 100324 to Zucker fa/fa rats reduces food intake and body weight gain, lowers plasma insulin concentrations while maintaining normoglycaemia, indicating an improvement of insulin sensitivity.

Asunto(s)

Hipoglucemiantes/farmacología , Resistencia a la Insulina , Ácidos Nicotínicos/farmacología , Obesidad/metabolismo , Receptores de Ácido Retinoico/agonistas , Tetrahidronaftalenos/farmacología , Tiazoles/farmacología , Tiazolidinedionas , Factores de Transcripción/agonistas , Análisis de Varianza , Animales , Glucemia/efectos de los fármacos , Peso Corporal/efectos de los fármacos , Ingestión de Alimentos/efectos de los fármacos , Insulina/sangre , Masculino , Ratas , Ratas Zucker , Receptores X Retinoide , Rosiglitazona

RESUMEN

Binding of agonists to nuclear receptors results in a conformational change in receptor structure that promotes interaction between activated receptors and coactivators. Receptor-coactivator interactions are mediated by the agonist-dependent formation of a hydrophobic pocket on the part of receptors, and short leucine-rich sequences termed LxxLL motifs or nuclear receptor boxes present in coactivators. RXR-PPARgamma (retinoid X receptor-peroxisome proliferator-activated receptor-gamma) heterodimers play important roles in adipocyte and macrophage differentiation and have been implicated as therapeutic targets in diabetes, atherosclerosis, and cancer. Analysis of interactions between RXR-PPARgamma heterodimers and coactivator nuclear receptor boxes suggests that RXR and PPARgamma can distinguish among coactivators by recognizing distinct structural features of nuclear receptor boxes. The results also indicate that coactivator choice by RXR is mediated by three nonconserved amino acids of the nuclear receptor box. The ability of an optimized seven-amino acid nuclear receptor box to specifically interact with RXR and function as a selective inhibitor suggests the coactivator-binding pocket may serve as a new target for drug discovery.

Asunto(s)

Receptores de Ácido Retinoico/genética , Receptores de Ácido Retinoico/metabolismo , Transactivadores/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Sustitución de Aminoácidos , Animales , Sitios de Unión , Proteína de Unión a CREB , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Línea Celular , Histona Acetiltransferasas , Subunidad 1 del Complejo Mediador , Datos de Secuencia Molecular , Mutación , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Coactivador 1 de Receptor Nuclear , Receptores Citoplasmáticos y Nucleares/genética , Receptores Citoplasmáticos y Nucleares/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Receptores X Retinoide , Transactivadores/genética

RESUMEN

The intracellular fatty acid content of insulin-sensitive target tissues determines in part their insulin sensitivity. Uptake of fatty acids into cells is a controlled process determined in part by a regulated import/export system that is controlled at least by two key groups of proteins, i.e. the fatty acid transport protein (FATP) and acyl-CoA synthetase (ACS), which facilitate, respectively, the transport of fatty acids across the cell membrane and catalyze their esterification to prevent their efflux. Previously it was shown that the expression of the FATP-1 and ACS genes was controlled by insulin and by peroxisome proliferator-activated receptor (PPAR) agonists in liver or in adipose tissue. The aim of this investigation was to determine the effects of retinoic acid derivatives on the expression of FATP-1 and ACS. In several cultured cell lines, it was shown that the expression of both the FATP-1 and ACS mRNAs was specifically induced at the transcriptional level by selective retinoid X receptor (RXR) but not by retinoic acid receptor (RAR) ligands. This effect was most pronounced in hepatoma cell lines. A similar induction of FATP-1 and ACS mRNA levels was also observed in vivo in Zucker diabetic fatty rats treated with the RXR agonist, LGD1069 (4-[1-(3,5,5,8,8-pentamethyl-5,6,7, 8-tetrahydro-2-naphthyl)ethenyl]benzoic acid). Through the use of heterodimer-selective compounds, it was demonstrated that the modulatory effect of these rexinoids on FATP-1 and ACS gene expression was mediated through activation of RXR in the context of the PPAR-RXR heterodimer. The observation that both RXR and PPAR agonists can stimulate the transcription of genes implicated in lipid metabolism, suggest that rexinoids may also act as lipid-modifying agents and support a role of the permissive PPAR-RXR heterodimer in the control of insulin sensitivity.

Asunto(s)

Proteínas Portadoras/metabolismo , Coenzima A Ligasas/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas de Transporte de Membrana , Receptores Citoplasmáticos y Nucleares/metabolismo , Receptores de Ácido Retinoico/metabolismo , Factores de Transcripción/metabolismo , Tretinoina/fisiología , Células 3T3 , Animales , Células CACO-2 , Proteínas Portadoras/genética , Núcleo Celular/metabolismo , Coenzima A Ligasas/genética , Dactinomicina/farmacología , Relación Dosis-Respuesta a Droga , Proteínas de Transporte de Ácidos Grasos , Proteínas de Unión a Ácidos Grasos , Ácidos Grasos/metabolismo , Humanos , Masculino , Proteínas de la Membrana/genética , Ratones , Inhibidores de la Síntesis del Ácido Nucleico/farmacología , ARN/metabolismo , Ratas , Ratas Zucker , Receptores Citoplasmáticos y Nucleares/química , Receptores de Ácido Retinoico/química , Receptores X Retinoide , Albúmina Sérica Bovina/metabolismo , Factores de Tiempo , Distribución Tisular , Factores de Transcripción/química , Células Tumorales Cultivadas

RESUMEN

Retinoids modulate the growth and differentiation of cancer cells presumably by activating gene transcription via the nuclear retinoic acid receptor (RAR) alpha, beta, and gamma and retinoid X receptor (RXR) alpha, beta, and gamma. We analyzed the effects of 38 RAR-selective and RXR-selective retinoids on the proliferation of 10 human head and neck squamous cell carcinoma (HNSCC) cell lines. All of these cell lines expressed constitutively all of the receptor subtypes except RARbeta, which was detected in only two of them. Most of the RAR-selective retinoids inhibited the growth of HNSCC cells to varying degrees, whereas the RXR-selective retinoids showed very weak or no inhibitory effects. Three RAR antagonists suppressed growth inhibition by RAR-selective agonists, as well as by RAR/RXR panagonists such as 9-cis-retinoic acid. Combinations of RXR-selective and RAR-selective retinoids exhibited additive growth-inhibitory effects. Furthermore, we found that CD437, the most potent growth-inhibitory retinoid induced apoptosis and up-regulated the expression of several apoptosis-related genes in HNSCC cells. These results indicate that: (a) retinoid receptors are involved in the growth-inhibitory effects of retinoids; (b) RXR-RAR heterodimers rather than RXR-RXR homodimer are the major mediators of growth inhibition by retinoids in HNSCC cells; and (c) induction of apoptosis can account for one mechanism by which retinoids such as CD437 inhibit the growth of HNSCC cells. Finally, these studies identified several synthetic retinoids, which are much more effective than the natural RAs and can be good candidates for chemoprevention and therapy of head and neck cancers.

Asunto(s)

Carcinoma de Células Escamosas/tratamiento farmacológico , División Celular/efectos de los fármacos , Neoplasias de Cabeza y Cuello/tratamiento farmacológico , Receptores de Ácido Retinoico/antagonistas & inhibidores , Retinoides/farmacología , Factores de Transcripción/antagonistas & inhibidores , Apoptosis/efectos de los fármacos , Apoptosis/genética , Northern Blotting , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patología , Análisis Mutacional de ADN , ADN de Neoplasias/química , ADN de Neoplasias/genética , Relación Dosis-Respuesta a Droga , Regulación Neoplásica de la Expresión Génica , Neoplasias de Cabeza y Cuello/genética , Neoplasias de Cabeza y Cuello/patología , Humanos , Mutación , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores de Ácido Retinoico/agonistas , Receptores de Ácido Retinoico/genética , Receptores X Retinoide , Factor de Transcripción AP-1/antagonistas & inhibidores , Factores de Transcripción/genética , Células Tumorales Cultivadas , Proteína p53 Supresora de Tumor/genética

Asunto(s)

Cara/fisiopatología , Hipoestesia/etiología , Sarcoidosis/complicaciones , Sarcoidosis/patología , Encéfalo/patología , Encéfalo/fisiopatología , Femenino , Humanos , Persona de Mediana Edad , Miositis/etiología , Miositis/patología , Enfermedades del Sistema Nervioso Periférico/etiología , Enfermedades del Sistema Nervioso Periférico/patología , Enfermedades del Sistema Nervioso Periférico/fisiopatología

RESUMEN

BACKGROUND: We have previously shown that a retinoid X receptor (RXR)-selective ligand (a rexinoid), called LGD1069, is highly efficacious in both the chemoprevention and the chemotherapy for N-nitrosomethylurea-induced rat mammary carcinomas. To evaluate a possible role for rexinoids in breast cancer therapy further, we have designed and characterized a novel carcinogen-induced model to mimic the clinical situation in which the tumors of patients stop responding to tamoxifen therapy and develop resistance to this drug. METHODS: Rats with experimentally induced mammary tumors were treated with tamoxifen to select a population with primary tumors that failed to respond completely to the drug. Once the failure of tamoxifen therapy had been established, LGD1069 was added to the treatment regimen, and the tumors in these animals were compared with tumors in a group of animals that remained on tamoxifen alone. RESULTS: LGD1069 in combination with tamoxifen for up to 20 weeks yielded an overall objective response rate of 94% (95% confidence interval [CI] = 86%-100%) (includes complete and partial responses) in primary tumors compared with a rate of 33% (95% CI = 11%-56%) in primary tumors treated with tamoxifen alone, a statistically significant difference (two-sided P<.0001). In addition, the LGD1069 and tamoxifen combination was associated with a statistically significant decrease in total tumor burden (two-sided P =.03). In a second study, tumors that failed to respond to tamoxifen therapy exhibited a 51% (95% CI = 34%-71%) objective response rate when treated with LGD1069 alone for 6 weeks after tamoxifen therapy was withdrawn. CONCLUSION: We have demonstrated that the RXR-selective ligand LGD1069 in combination with tamoxifen is a highly efficacious therapeutic agent for tumors that fail to respond completely to tamoxifen. This finding suggests that rexinoid therapy offers a novel approach to the treatment of breast tumors that may have developed resistance to antihormonal therapies such as tamoxifen.

Asunto(s)

Anticarcinógenos/farmacología , Antineoplásicos Hormonales/uso terapéutico , Neoplasias Mamarias Experimentales/tratamiento farmacológico , Tamoxifeno/uso terapéutico , Tetrahidronaftalenos/farmacología , Animales , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Bexaroteno , Carcinógenos , Progresión de la Enfermedad , Femenino , Neoplasias Mamarias Experimentales/inducido químicamente , Metilnitrosourea , Ratas , Ratas Sprague-Dawley , Insuficiencia del Tratamiento , Resultado del Tratamiento

RESUMEN

BACKGROUND: The secosteroid 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) acts through the vitamin D receptor (VDR) to elicit many activities that make it a promising drug candidate for the treatment of a number of diseases, including cancer and psoriasis. Clinical use of 1,25(OH)2D3 has been limited by hypercalcemia elicited by pharmacologically effective doses. We hypothesized that structurally distinct, nonsecosteroidal mimics of 1,25(OH)2D3 might have different activity profiles from vitamin D analogs, and set out to discover such compounds by screening small-molecule libraries. RESULTS: A bis-phenyl derivative was found to activate VDR in a transactivation screening assay. Additional related compounds were synthesized that mimicked various activities of 1,25(OH)2D3, including growth inhibition of cancer cells and keratinocytes, as well as induction of leukemic cell differentiation. In contrast to 1, 25(OH)2D3, these synthetic compounds did not demonstrate appreciable binding to serum vitamin D binding protein, a property that is correlated with fewer calcium effects in vivo. Two mimics tested in mice showed greater induction of a VDR target gene with less elevation of serum calcium than 1,25(OH)2D3. CONCLUSIONS: These novel VDR modulators may have potential as therapeutics for cancer, leukemia and psoriasis with less calcium mobilization side effects than are associated with secosteroidal 1,25(OH)2D3 analogs.

Asunto(s)

Antineoplásicos/farmacología , Calcio/metabolismo , Receptores de Calcitriol/fisiología , Vitamina D/farmacología , Animales , Transporte Biológico , Neoplasias de la Mama/patología , Calcitriol/farmacología , Agonistas de los Canales de Calcio/farmacología , Diferenciación Celular/efectos de los fármacos , División Celular/efectos de los fármacos , Femenino , Células HL-60 , Humanos , Queratinocitos/citología , Queratinocitos/efectos de los fármacos , Cetonas/farmacología , Macrófagos/citología , Macrófagos/efectos de los fármacos , Masculino , Ratones , Imitación Molecular , Éteres Fenílicos/farmacología , Neoplasias de la Próstata/patología , Ratas , Receptores de Calcitriol/metabolismo , Activación Transcripcional , Vitamina D/análogos & derivados , Vitamina D/síntesis química , Proteína de Unión a Vitamina D/metabolismo

RESUMEN

The multiple biologic activities of retinoic acid (RA) are mediated through RAR and retinoid X receptor (RXR) nuclear receptors that interact with specific DNA target sequences as heterodimers (RXR-RAR) or homodimers (RXR-RXR). RA receptor activation appears critical to regulating important aspects of hematopoiesis, since transducing a COOH-terminally truncated RARalpha exhibiting dominant-negative activity (RARalpha403) into normal mouse bone marrow generates hematopoietic growth factor-dependent cell lines frozen at the multipotent progenitor (EML) or committed promyelocyte (MPRO) stages. Nevertheless, relatively high, pharmacological concentrations of RA (1 to 10 microM) overcome these differentiation blocks and induce terminal granulocytic differentiation of the MPRO promyelocytes while potentiating interleukin-3 (IL-3)-induced commitment of EML cells to the granulocyte/monocyte lineage. In the present study, we utilized RXR- and RAR-specific agonists and antagonists to determine how RA overcomes the dominant-negative activity of the truncated RARalpha in these different myeloid developmental stages. Unexpectedly, we observed that an RXR-specific, rather than an RAR-specific, agonist induces terminal granulocytic differentiation of MPRO promyelocytes, and this differentiation is associated with activation of DNA response elements corresponding to RAR-RXR heterodimers rather than RXR-RXR homodimers. This RXR agonist activity is blocked by RAR-specific antagonists, suggesting extensive cross-talk between the partners of the RXR-RARalpha403 heterodimer. In contrast, in the more immature, multipotent EML cells we observed that this RXR-specific agonist is inactive either in potentiating IL-3-mediated commitment of EML cells to the granulocyte lineage or in transactivating RAR-RXR response elements. RA-triggered GALdbd-RARalpha hybrid activity in these cells indicates that the multipotent EML cells harbor substantial nuclear hormone receptor coactivator activity. However, the histone deacetylase (HDAC) inhibitor trichostatin A readily activates an RXR-RAR reporter construct in the multipotent EML cells but not in the committed MPRO promyelocytes, indicating that differences in HDAC-containing repressor complexes in these two closely related but distinct hematopoietic lineages might account for the differential activation of the RXR-RARalpha403 heterodimers that we observed at these different stages of myeloid development.

Asunto(s)

Diferenciación Celular/genética , Regulación del Desarrollo de la Expresión Génica/genética , Granulocitos/metabolismo , Receptores de Ácido Retinoico/agonistas , Factores de Transcripción/agonistas , Dimerización , Genes Reporteros , Histona Desacetilasas/metabolismo , Humanos , Ácidos Hidroxámicos/farmacología , Interleucina-3/farmacología , Proteínas Nucleares/análisis , Oligodesoxirribonucleótidos/genética , Oligodesoxirribonucleótidos/metabolismo , ARN Mensajero/metabolismo , Receptores de Superficie Celular/genética , Receptores de Ácido Retinoico/antagonistas & inhibidores , Receptores X Retinoide , Retinoides/farmacología , Células Madre/metabolismo , Factores de Transcripción/antagonistas & inhibidores , Activación Transcripcional/efectos de los fármacos , Tretinoina/farmacología

RESUMEN

A novel series of oxime ligands has been synthesized that displays potent, specific activation of the retinoid X receptors (RXRs). The oximes of 3-substituted (tetramethyltetrahydronaphthyl)carbonylbenzoic acids are readily available by condensation with hydroxyl- or methoxylamine; alkylation of the hydroxyl oxime provides a variety of analogues. Oximes and variously substituted oxime derivatives demonstrate high binding affinity for the RXRs and specific RXR activation and, hence, are called rexinoids. These oxime rexinoids are activators of the RXR:PPARgamma heterodimer and are potent inducers of differentiation of 3T3-L1 preadipocytes to adipocytes. We have recently reported that ligands which activate the RXR:PPARgamma heterodimer in this manner are effective in the treatment of type II diabetes (non-insulin-dependent diabetes mellitus, NIDDM). Thus, these new oxime rexinoids are potential therapeutic agents for the treatment of metabolic disorders, such as obesity and diabetes.

Asunto(s)

Adipocitos/efectos de los fármacos , Benzoatos/síntesis química , Proteínas de Unión al ADN/metabolismo , Oximas/síntesis química , Receptores de Ácido Retinoico/agonistas , Factores de Transcripción/agonistas , Células 3T3 , Adipocitos/metabolismo , Adipocitos/fisiología , Animales , Benzoatos/química , Benzoatos/metabolismo , Benzoatos/farmacología , Unión Competitiva , Diferenciación Celular/efectos de los fármacos , Línea Celular , Cristalografía por Rayos X , Ligandos , Ratones , Oximas/química , Oximas/metabolismo , Oximas/farmacología , Receptores Citoplasmáticos y Nucleares/agonistas , Receptores Citoplasmáticos y Nucleares/biosíntesis , Receptores de Ácido Retinoico/biosíntesis , Receptores de Ácido Retinoico/metabolismo , Proteínas Recombinantes/agonistas , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/metabolismo , Receptores X Retinoide , Factores de Transcripción/biosíntesis , Factores de Transcripción/metabolismo , Transfección , Triglicéridos/metabolismo

RESUMEN

Retinoids regulate gene expression through the action of retinoic acid receptors (RARs) and retinoid-X receptors (RXRs), which both belong to the family of nuclear hormone receptors. Retinoids are of fundamental importance during development, but it has been difficult to assess the distribution of ligand-activated receptors in vivo. This is particularly the case for RXR, which is a critical unliganded auxiliary protein for several nuclear receptors, including RAR, but its ligand-activated role in vivo remains uncertain. Here we describe an assay in transgenic mice, based on the expression of an effector fusion protein linking the ligand-binding domain of either RXR or RAR to the yeast Gal4 DNA-binding domain, and the in situ detection of ligand-activated effector proteins by using an inducible transgenic lacZ reporter gene. We detect receptor activation in the spinal cord in a pattern that indicates that the receptor functions in the maturation of limb-innervating motor neurons. Our results reveal a specific activation pattern of Gal4-RXR which indicates that RXR is a critical bona fide receptor in the developing spinal cord.

Asunto(s)

Receptores de Ácido Retinoico/metabolismo , Transducción de Señal , Médula Espinal/embriología , Factores de Transcripción/metabolismo , Animales , Técnicas de Cultivo , Extremidades/embriología , Extremidades/inervación , Genes Reporteros , Humanos , Ratones , Ratones Transgénicos , Neuronas Motoras/fisiología , Receptores X Retinoide , Médula Espinal/metabolismo , Células Tumorales Cultivadas , beta-Galactosidasa

RESUMEN

The ability of DNA sequence-specific transcription factors to synergistically activate transcription is a common property of genes transcribed by RNA polymerase II. The present work characterizes a unique form of intermolecular transcriptional synergy between two members of the nuclear hormone receptor superfamily. Heterodimers formed between peroxisome proliferator-activated receptor gamma (PPARgamma), an adipocyte-enriched member of the superfamily required for adipogenesis, and retinoid X receptors (RXRs) can activate transcription in response to ligands specific for either subunit of the dimer. Simultaneous treatment with ligands specific for both PPARgamma and RXR has a synergistic effect on the transactivation of reporter genes and on adipocyte differentiation in cultured cells. Mutation of the PPARgamma hormone-dependent activation domain (named tauc or AF-2) inhibits the ability of RXR-PPARgamma heterodimers to respond to ligands specific for either subunit. In contrast, the ability of RXR- and PPARgamma-specific ligands to synergize does not require the hormone-dependent activation domain of RXR. The results of in vitro and in vivo experiments indicate that binding of ligands to RXR alters the conformation of the dimerization partner, PPARgamma, and modulates the activity of the heterodimer in a manner independent of the RXR hormone-dependent activation domain.

Asunto(s)

Proteínas de Unión al ADN/metabolismo , Microcuerpos/metabolismo , Proteínas Nucleares/metabolismo , Receptores Citoplasmáticos y Nucleares/metabolismo , Receptores de Ácido Retinoico/metabolismo , Factores de Transcripción/metabolismo , Activación Transcripcional , Células 3T3 , Adipocitos/citología , Animales , Sitios de Unión , Diferenciación Celular , Dimerización , Histona Acetiltransferasas , Ligandos , Sustancias Macromoleculares , Ratones , Coactivador 1 de Receptor Nuclear , Conformación Proteica , Receptores X Retinoide

RESUMEN

Peroxisome proliferator-activated receptors (PPARs) and retinoid X receptors (RXRs) are members of the intracellular receptor superfamily. PPARs bind to peroxisome proliferator-response elements (PPREs) as heterodimers with RXR and as such activate gene transcription in response to activators. Fibrates like gemfibrozil are well-known PPARalpha activators and are used in the treatment of hyperlipidemia. We show that the RXR ligand LGD1069 (Targretin), like gemfibrozil, can activate the PPARalpha/RXR signal-transduction pathway, including transactivation of the bifunctional enzyme or acyl-CoA oxidase response elements in a cotransfection assay. The activation also occurs in vivo, whereby in rats treated with LGD1069 or gemfibrozil, bifunctional enzyme and acyl-CoA oxidase RNA are induced and the combination of LGD1069 and gemfibrozil leads to a greater induction. Importantly, in hypertriglyceridemic db/db mice treated with RXR or PPARalpha agonists, triglyceride levels are lowered, and the combination again has significantly greater efficacy. RXR agonists also raise HDL cholesterol levels without changing apoA-I RNA expression. This observation suggests the use of RXR-selective agonists, "rexinoids," either alone or in combination with a fibrate as a new therapeutic approach to treating patients with high triglyceride and low HDL cholesterol levels.

Asunto(s)

HDL-Colesterol/sangre , Regulación de la Expresión Génica/fisiología , Receptores Citoplasmáticos y Nucleares/fisiología , Receptores de Ácido Retinoico/agonistas , Factores de Transcripción/agonistas , Factores de Transcripción/fisiología , Triglicéridos/sangre , Acil-CoA Oxidasa , Animales , Bexaroteno , Combinación de Medicamentos , Femenino , Gemfibrozilo/farmacología , Hipertrigliceridemia/sangre , Hipertrigliceridemia/genética , Hipolipemiantes/farmacología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Mutantes , Oxidorreductasas/genética , Ratas , Ratas Sprague-Dawley , Receptores X Retinoide , Transducción de Señal/efectos de los fármacos , Tetrahidronaftalenos/farmacología , Activación Transcripcional/genética

RESUMEN

Recently, we reported that LGD1069, a high-affinity ligand for the retinoid X receptors (RXRs), was shown to have an efficacy equivalent to that of tamoxifen (TAM) as a chemopreventive agent in the N-nitroso-N-methylurea-induced rat mammary carcinoma model. Furthermore, LGD1069 was very well tolerated during 13 weeks of chronic therapy with no classic signs of "retinoid-associated" toxicities. Due to the high efficacy and benign profile of this RXR agonist as a suppressor of carcinogenesis, we examined its role as a therapeutic agent on established mammary carcinomas. In the rat mammary carcinoma model, N-nitroso-N-methylurea was used to induce tumors, and the tumors were allowed to grow to an established size prior to initiation of treatment. LGD1069-treated animals showed complete regression in 72% of treated tumors and had a reduced tumor load compared to control. In addition, the combination of LGD1069 and TAM showed increased efficacy over either agent alone. Histopathological analysis showed a reduction of LGD1069-treated tumor malignancy, an increase in differentiation, and a sharp decrease in cellular proliferation compared to vehicle-treated control tumors. These data demonstrate that the RXR-selective ligand LGD1069 is a highly efficacious therapeutic agent for mammary carcinoma and enhances the activity of TAM.

Asunto(s)

Antineoplásicos/uso terapéutico , Neoplasias Mamarias Experimentales/tratamiento farmacológico , Receptores de Ácido Retinoico/agonistas , Tetrahidronaftalenos/uso terapéutico , Factores de Transcripción/agonistas , Animales , Anticarcinógenos/uso terapéutico , Antineoplásicos Hormonales/administración & dosificación , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Bexaroteno , División Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Sinergismo Farmacológico , Femenino , Neoplasias Mamarias Experimentales/inducido químicamente , Neoplasias Mamarias Experimentales/patología , Metilnitrosourea , Ratas , Ratas Sprague-Dawley , Receptores X Retinoide , Tamoxifeno/administración & dosificación , Tetrahidronaftalenos/administración & dosificación

RESUMEN

Vitamin A and its derivatives (collectively referred to as retinoids) are required for many fundamental life processes, including vision, reproduction, metabolism, cellular differentiation, hematopoesis, bone development, and pattern formation during embryogenesis. There is also considerable evidence to suggest that natural and synthetic retinoids have therapeutical effects due to their antiproliferative and apoptosis-inducing effects in human diseases such as cancer. Therefore it is not surprising that a significant amount of research was dedicated to probe the molecular and cellular mechanisms of retinoid action during the past decade. One of the cellular mechanisms retinoids have been implicated in is the initiation and modulation of apoptosis in normal development and disease. This review provides a brief overview of the molecular basis of retinoid signaling, and focuses on the retinoid-regulation of apoptotic cell death and gene expression during normal development and in pathological conditions in vivo and in various tumor cell lines in vitro.

Asunto(s)

Apoptosis/fisiología , Regulación Neoplásica de la Expresión Génica , Neoplasias/fisiopatología , Receptores de Ácido Retinoico/genética , Tretinoina/fisiología , Humanos , Neoplasias/genética