Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 38
Filtrar
Más filtros

Base de datos
Tipo del documento
Intervalo de año de publicación
1.
Int J Mol Sci ; 22(3)2021 Jan 26.
Artículo en Inglés | MEDLINE | ID: mdl-33530349

RESUMEN

Tauopathies are a group of more than twenty known disorders that involve progressive neurodegeneration, cognitive decline and pathological tau accumulation. Current therapeutic strategies provide only limited, late-stage symptomatic treatment. This is partly due to lack of understanding of the molecular mechanisms linking tau and cellular dysfunction, especially during the early stages of disease progression. In this study, we treated early stage tau transgenic mice with a multi-target kinase inhibitor to identify novel substrates that contribute to cognitive impairment and exhibit therapeutic potential. Drug treatment significantly ameliorated brain atrophy and cognitive function as determined by behavioral testing and a sensitive imaging technique called manganese-enhanced magnetic resonance imaging (MEMRI) with quantitative R1 mapping. Surprisingly, these benefits occurred despite unchanged hyperphosphorylated tau levels. To elucidate the mechanism behind these improved cognitive outcomes, we performed quantitative proteomics to determine the altered protein network during this early stage in tauopathy and compare this model with the human Alzheimer's disease (AD) proteome. We identified a cluster of preserved pathways shared with human tauopathy with striking potential for broad multi-target kinase intervention. We further report high confidence candidate proteins as novel therapeutically relevant targets for the treatment of tauopathy. Proteomics data are available via ProteomeXchange with identifier PXD023562.


Asunto(s)
Neuronas/efectos de los fármacos , Neuronas/metabolismo , Inhibidores de Proteínas Quinasas/farmacología , Tauopatías/etiología , Tauopatías/metabolismo , Animales , Encéfalo/metabolismo , Encéfalo/patología , Modelos Animales de Enfermedad , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Hipocampo/patología , Humanos , Ratones , Ratones Transgénicos , Enfermedades Neurodegenerativas/diagnóstico , Enfermedades Neurodegenerativas/tratamiento farmacológico , Enfermedades Neurodegenerativas/etiología , Enfermedades Neurodegenerativas/metabolismo , Fosforilación , Inhibidores de Proteínas Quinasas/uso terapéutico , Proteoma , Proteómica/métodos , Índice de Severidad de la Enfermedad , Tauopatías/diagnóstico , Tauopatías/tratamiento farmacológico , Respuesta de Proteína Desplegada , eIF-2 Quinasa/metabolismo , Proteínas tau/metabolismo
2.
Nat Commun ; 10(1): 4659, 2019 10 11.
Artículo en Inglés | MEDLINE | ID: mdl-31604915

RESUMEN

MG53 is a muscle-specific TRIM-family protein that presides over the cell membrane repair response. Here, we show that MG53 present in blood circulation acts as a myokine to facilitate tissue injury-repair and regeneration. Transgenic mice with sustained elevation of MG53 in the bloodstream (tPA-MG53) have a healthier and longer life-span when compared with littermate wild type mice. The tPA-MG53 mice show normal glucose handling and insulin signaling in skeletal muscle, and sustained elevation of MG53 in the bloodstream does not have a deleterious impact on db/db mice. More importantly, the tPA-MG53 mice display remarkable dermal wound healing capacity, enhanced muscle performance, and improved injury-repair and regeneration. Recombinant human MG53 protein protects against eccentric contraction-induced acute and chronic muscle injury in mice. Our findings highlight the myokine function of MG53 in tissue protection and present MG53 as an attractive biological reagent for regenerative medicine without interference with glucose handling in the body.


Asunto(s)
Proteínas de la Membrana/fisiología , Cicatrización de Heridas , Animales , Calcio/metabolismo , Glucosa/metabolismo , Prueba de Tolerancia a la Glucosa , Insulina/metabolismo , Proteínas de la Membrana/sangre , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Músculo Esquelético/metabolismo , Regeneración/genética , Biología de Sistemas
3.
Hum Mol Genet ; 27(14): 2477-2489, 2018 07 15.
Artículo en Inglés | MEDLINE | ID: mdl-29718201

RESUMEN

Marinesco-Sjögren syndrome (MSS) is a rare, early onset, autosomal recessive multisystem disorder characterized by cerebellar ataxia, cataracts and myopathy. Most MSS cases are caused by loss-of-function mutations in the gene encoding SIL1, a nucleotide exchange factor for the molecular chaperone BiP which is essential for correct protein folding in the endoplasmic reticulum. Woozy mice carrying a spontaneous Sil1 mutation recapitulate key pathological features of MSS, including cerebellar atrophy with degeneration of Purkinje cells and progressive myopathy. Because the PERK branch of the unfolded protein response is activated in degenerating neurons of woozy mice, and inhibiting PERK-mediated translational attenuation has shown protective effects in protein-misfolding neurodegenerative disease models, we tested the therapeutic efficacy of GSK2606414, a potent inhibitor of PERK. Mice were chronically treated with GSK2606414 starting from a presymptomatic stage, and the effects were evaluated on biochemical, histopathological and clinical readouts. GSK2606414 delayed Purkinje cell degeneration and the onset of motor deficits, prolonging the asymptomatic phase of the disease; it also reduced the skeletal muscle abnormalities and improved motor performance during the symptomatic phase. The protein but not the mRNA level of ORP150, a nucleotide exchange factor which can substitute for SIL1, was increased in the cerebellum of GSK2606414-treated woozy mice, suggesting that translational recovery promoted the synthesis of this alternative BiP co-factor. Targeting PERK signaling may have beneficial disease-modifying effects in carriers of SIL1 mutations.


Asunto(s)
Factores de Intercambio de Guanina Nucleótido/genética , Proteínas HSP70 de Choque Térmico/genética , Degeneración Nerviosa/genética , Degeneraciones Espinocerebelosas/terapia , eIF-2 Quinasa/genética , Adenina/administración & dosificación , Adenina/análogos & derivados , Animales , Cerebelo/efectos de los fármacos , Cerebelo/fisiopatología , Modelos Animales de Enfermedad , Retículo Endoplásmico/genética , Retículo Endoplásmico/patología , Heterocigoto , Humanos , Indoles/administración & dosificación , Mutación con Pérdida de Función/genética , Ratones , Actividad Motora/fisiología , Degeneración Nerviosa/fisiopatología , Pliegue de Proteína , Células de Purkinje/efectos de los fármacos , Células de Purkinje/patología , Degeneraciones Espinocerebelosas/genética , Degeneraciones Espinocerebelosas/patología , Respuesta de Proteína Desplegada/genética
4.
Blood Adv ; 1(26): 2553-2562, 2017 Dec 12.
Artículo en Inglés | MEDLINE | ID: mdl-29296907

RESUMEN

Neutropenia is a common consequence of radiation and chemotherapy in cancer patients. The resulting immunocompromised patients become highly susceptible to potentially life-threatening infections. Granulocyte colony-stimulating factor (G-CSF) is known to stimulate neutrophil production and is widely used as a treatment of chemotherapy-induced neutropenia. A small-molecule G-CSF secretagogue without a requirement for refrigerated supply chain would offer a more convenient and cost-effective treatment of chemotherapy-induced neutropenia. Bacterial lipopeptides activate innate immune responses through Toll-like receptor 2 (TLR2) and induce the release of cytokines, including G-CSF, from macrophages, monocytes, and endothelial. Pam2CSK4 is a synthetic lipopeptide that effectively mimics bacterial lipoproteins known to activate TLR2 receptor signaling through the TLR2/6 heterodimer. Substrate-based drug design led to the discovery of GSK3277329, which stimulated the release of G-CSF in activated THP-1 cells, peripheral blood mononuclear cells, and human umbilical vein endothelial cells. When administered subcutaneously to cynomolgus monkeys (Macaca fascicularis), GSK3277329 caused systemic elevation of G-CSF and interleukin-6 (IL-6), but not IL-1ß or tumor necrosis factor α, indicating a selective cytokine-stimulation profile. Repeat daily injections of GSK3277329 in healthy monkeys also raised circulating neutrophils above the normal range over a 1-week treatment period. More importantly, repeated daily injections of GSK3277329 over a 2-week period restored neutrophil loss in monkeys given chemotherapy treatment (cyclophosphamide, Cytoxan). These data demonstrate preclinical in vivo proof of concept that TLR2 agonism can drive both G-CSF induction and subsequent neutrophil elevation in the cynomolgus monkey and could be a therapeutic strategy for the treatment of chemotherapy-induced neutropenia.

5.
Int J Urol ; 21(3): 319-24, 2014 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-24033563

RESUMEN

OBJECTIVES: To study the relationship between myosin light chain phosphorylation of the detrusor muscle and spontaneous smooth muscle contractions in a rabbit model of partial outlet obstruction. METHODS: New Zealand white rabbit urinary bladders were partially obstructed for 2 weeks. Rabbits were euthanized, detrusor muscle strips were hung on a force transducer and spontaneous activity was measured at varying concentrations (0-0.03 µM/L) of the Rho-kinase inhibitors GSK 576371 or 0.01 µM/L Y27632. Basal myosin light chain phosphorylation was measured by 2-D gel electrophoresis in control and GSK 576371-treated strips. RESULTS: Both drugs suppressed the force of spontaneous contractions, whereas GSK 576371 had a more profound effect on the frequency of the contractions. The IC50 values for the inhibition of frequency and force of spontaneous contractions were 0.17 µM/L and 0.023 µM/L for GSK 576371, respectively. The compound significantly decreased the basal myosin light chain phosphorylation from 28.0 ± 3.9% to 13.5 ± 1.9% (P < 0.05). At 0.01 µM/L, GSK 576371 inhibited spontaneous bladder overactivity by 50%, but inhibited carbachol-elicited contractions force by just 25%. CONCLUSIONS: These data suggest that Rho-kinase regulation of myosin light chain phosphorylation contributes to the spontaneous detrusor activity induced by obstruction. This finding could have therapeutic implications by providing another therapeutic option for myogenic, overactive bladder.


Asunto(s)
Inhibidores Enzimáticos/farmacología , Cadenas Ligeras de Miosina/metabolismo , Vejiga Urinaria Hiperactiva/metabolismo , Quinasas Asociadas a rho/antagonistas & inhibidores , Animales , Masculino , Datos de Secuencia Molecular , Fosforilación/efectos de los fármacos , Conejos , Obstrucción del Cuello de la Vejiga Urinaria/complicaciones , Vejiga Urinaria Hiperactiva/etiología
6.
Urology ; 79(6): 1410.e1-6, 2012 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-22497980

RESUMEN

OBJECTIVE: To evaluate the consequence of pharmacologic inhibition of voltage-gated Na(+) channels (Nav) in the conscious rat, based on Nav having been implicated as modulators of rodent urodynamics using knockout as well as antisense oligodeoxynucleotide approaches. METHODS: The urodynamic response to standard Nav blockers, lamotrigine, amitriptyline, mexiletine, and carbamazepine were evaluated using conscious, continuous-filling cystometry in spontaneously hypertensive rats (SHRs). As a selectivity evaluation, the activity of the Nav blockers at muscarinic receptors was assessed via effect on carbachol-evoked bladder contractions. RESULTS: Lamotrigine, amitriptyline, mexiletine, and carbamazepine decreased peak micturition pressure, micturition interval, and void volume. These effects were markedly similar to observations with muscarinic antagonists. Therefore, we evaluated the selectivity of these agents against bladder muscarinic receptors. Lamotrigine, mexiletine, and carbamazepine had no effect on muscarinic bladder contractions, whereas amitriptyline displayed a robust antagonism of carbachol-induced contractility. CONCLUSION: Three Nav blockers--lamotrigine, mexiletine, and carbamazepine--demonstrated a reduction in micturition pressure and functional bladder capacity, similar to previous observations with muscarinic antagonists. These 3 Nav blockers are free of muscarinic antagonism, consistent with their cystometric effects being mediated via their Nav blocking activities. The negative findings reported here with Nav blockers suggest that Nav channel blockade is unlikely to reflect an improved treatment strategy for bladder disorders over currently prescribed muscarinic antagonists.


Asunto(s)
Bloqueadores de los Canales de Sodio/farmacología , Canales de Sodio/efectos de los fármacos , Vejiga Urinaria Hiperactiva/tratamiento farmacológico , Amitriptilina/farmacología , Animales , Carbamazepina/farmacología , Femenino , Lamotrigina , Mexiletine/farmacología , Ratas , Ratas Endogámicas SHR , Receptores Muscarínicos/efectos de los fármacos , Triazinas/farmacología , Urodinámica/efectos de los fármacos
7.
Bioorg Med Chem Lett ; 20(1): 371-4, 2010 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-19926282

RESUMEN

The previously reported pyrrolidine class of progesterone receptor partial agonists demonstrated excellent potency but suffered from serious liabilities including hERG blockade and high volume of distribution in the rat. The basic pyrrolidine amine was intentionally converted to a sulfonamide, carbamate, or amide to address these liabilities. The evaluation of the degree of partial agonism for these non-basic pyrrolidine derivatives and demonstration of their efficacy in an in vivo model of endometriosis is disclosed herein.


Asunto(s)
Pirrolidinas/química , Receptores de Progesterona/agonistas , Animales , Sitios de Unión , Carbamatos/química , Cristalografía por Rayos X , Canal de Potasio ERG1 , Endometriosis/tratamiento farmacológico , Canales de Potasio Éter-A-Go-Go/metabolismo , Femenino , Humanos , Pirrolidinas/síntesis química , Pirrolidinas/farmacocinética , Ratas , Receptores de Progesterona/metabolismo , Sulfonamidas/química
8.
Steroids ; 74(13-14): 1015-24, 2009.
Artículo en Inglés | MEDLINE | ID: mdl-19665469

RESUMEN

Endometriosis, defined as the presence of endometrial glands and stroma at extra-uterine sites, is a gynecological condition that affects women of reproductive age. Consistent with its uterine origins, endometriotic lesions and resulting symptoms are hormonally responsive. To investigate Progesterone Receptor (PR)-based therapies, we measured physiological endpoints and gene expression in rat models of uterine cell estrogenic activity. Estrogen-induced ELT-3 rat leiomyoma cell proliferation was significantly inhibited by progesterone (P4), while the antiprogestin RU486 or the Selective PR Modulator (SPRM) asoprisnil, did not block proliferation. Stromal cell-derived factor-1 (SDF-1/Cxcl12) gene expression was induced by estrogen, and was repressed by the Selective Estrogen Receptor Modulators (SERMs), the antiestrogen ICI 182,780, and P4, but not by RU486 or the ERbeta-selective ligand ERB-041. In ELT-3 cells, asoprisnil demonstrated partial PR agonism on SDF-1 gene repression. Magnetic Resonance Imaging was used to monitor development of ectopic cysts in a rat surgical model of endometriosis. SERMs and P4 significantly decreased cyst volumes comparably by approximately 60%. However, ERB-041 and asoprisnil had no effect on cyst volume, and RU486 increased cyst volume by 20%. SDF-1 expression was modestly, but significantly, increased in the cyst compared to eutopic uterus, and P4 and raloxifene could repress the expression. We showed that SDF-1 was similarly regulated in human cells. These data suggest that transcriptional regulation of SDF-1 is a surrogate marker of estrogenic activities via ERalpha in rat uterine cells, and that SDF-1 repression by PR agonists can predict the ability to oppose the actions of estrogen in vivo.


Asunto(s)
Quimiocina CXCL12/antagonistas & inhibidores , Endometriosis/tratamiento farmacológico , Progesterona/uso terapéutico , Progestinas/uso terapéutico , Receptores de Progesterona/agonistas , Moduladores Selectivos de los Receptores de Estrógeno/uso terapéutico , Útero/efectos de los fármacos , Animales , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Quimiocina CXCL12/agonistas , Quimiocina CXCL12/metabolismo , Quistes/tratamiento farmacológico , Quistes/metabolismo , Quistes/patología , Modelos Animales de Enfermedad , Endometriosis/metabolismo , Endometriosis/patología , Estradiol/análogos & derivados , Estradiol/farmacología , Estrenos/farmacología , Antagonistas de Estrógenos/farmacología , Receptor alfa de Estrógeno/metabolismo , Estrógenos/farmacología , Femenino , Fulvestrant , Expresión Génica , Antagonistas de Hormonas/farmacología , Humanos , Mifepristona/farmacología , Oxazoles/farmacología , Oximas/farmacología , Progesterona/farmacología , Progestinas/farmacología , Ratas , Moduladores Selectivos de los Receptores de Estrógeno/farmacología , Útero/metabolismo
9.
Urology ; 74(2): 463-6, 2009 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-19573897

RESUMEN

OBJECTIVES: To examine the effects of estrogen and/or progesterone on the cystometric profiles obtained using continuous-filling cystometry in the conscious Sprague-Dawley rat. METHODS: Sprague-Dawley rats underwent ovariectomy (OVX) and were compared with controls by conscious continuous-filling cystometry. The effect of estrogen (10 microg/kg/d for 14 days) and/or progesterone (10 mg/kg/d for 14 days) replacement on OVX urodynamics was examined (n = 7-8/group). RESULTS: OVX rats demonstrated reduced micturition intervals and voided volumes compared with controls. These effects of OVX were reversed by estrogen replacement, but not by progesterone replacement. When combined with estrogen, progesterone functioned to partially antagonize the effects of estrogen in OVX rats. CONCLUSIONS: Estrogen enhances bladder capacity in the OVX rat and therefore is a likely contributor to the larger bladder capacity in the female compared with the male rat. Consistent with its established role in reproductive physiology, progesterone antagonizes the beneficial effects of estrogen on OVX rat urodynamics.


Asunto(s)
Estrógenos/fisiología , Progesterona/fisiología , Urodinámica/fisiología , Animales , Estrógenos/farmacología , Femenino , Ovariectomía , Progesterona/farmacología , Ratas , Ratas Sprague-Dawley , Vejiga Urinaria/fisiología , Urodinámica/efectos de los fármacos
10.
Bioorg Med Chem Lett ; 19(16): 4777-80, 2009 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-19595590

RESUMEN

Using the X-ray crystal structure of an amide-based progesterone receptor (PR) partial agonist bound to the PR ligand binding domain, a novel PR partial agonist class containing a pyrrolidine ring was designed. Members of this class of N-alkylpyrrolidines demonstrate potent and highly selective partial agonism of the progesterone receptor, and one of these analogs was shown to be efficacious upon oral dosing in the OVX rat model of estrogen opposition.


Asunto(s)
Pirrolidinas/química , Receptores de Progesterona/agonistas , Administración Oral , Animales , Sitios de Unión , Simulación por Computador , Cristalografía por Rayos X , Diseño de Fármacos , Modelos Animales , Estructura Terciaria de Proteína , Pirrolidinas/administración & dosificación , Pirrolidinas/síntesis química , Ratas , Receptores de Progesterona/metabolismo
11.
Bioorg Med Chem Lett ; 19(15): 4441-5, 2009 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-19497745

RESUMEN

The lead serum and glucocorticoid-related kinase 1 (SGK1) inhibitors 4-(5-phenyl-1H-pyrrolo[2,3-b]pyridin-3-yl)benzoic acid (1) and {4-[5-(2-naphthalenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]phenyl}acetic acid (2) suffer from low DNAUC values in rat, due in part to formation and excretion of glucuronic acid conjugates. These PK/glucuronidation issues were addressed either by incorporating a substituent on the 3-phenyl ring ortho to the key carboxylate functionality of 1 or by substituting on the group in between the carboxylate and phenyl ring of 2. Three of these analogs have been identified as having good SGK1 inhibition potency and have DNAUC values suitable for in vivo testing.


Asunto(s)
Química Farmacéutica/métodos , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/farmacología , Proteínas Inmediatas-Precoces/antagonistas & inhibidores , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Administración Oral , Animales , Disponibilidad Biológica , Diseño de Fármacos , Glucocorticoides/química , Ácido Glucurónico/química , Proteínas Inmediatas-Precoces/química , Concentración 50 Inhibidora , Modelos Químicos , Conformación Molecular , Inhibidores de Proteínas Quinasas/antagonistas & inhibidores , Proteínas Serina-Treonina Quinasas/química , Ratas , Relación Estructura-Actividad
12.
Bioorg Med Chem Lett ; 19(10): 2637-41, 2009 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-19376703

RESUMEN

Two classes of amino acid-derived heterocyclic progesterone receptor ligands were developed to address the metabolic issues posed by the dimethyl amide functionality of the lead compound (1). The tetrazole-derived ligands behaved as potent partial agonists, while the 1,2,4-triazole ligands behaved as potent full agonists.


Asunto(s)
Receptores de Progesterona/agonistas , Tetrazoles/síntesis química , Aminoácidos/química , Animales , Ratas , Receptores de Progesterona/metabolismo , Relación Estructura-Actividad , Tetrazoles/química , Tetrazoles/farmacocinética
13.
J Urol ; 181(3): 1444-51, 2009 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-19157444

RESUMEN

PURPOSE: Urodynamics have been traditionally recorded in anesthetized or conscious animals implanted with a bladder catheter that is used to artificially fill the bladder while measuring intravesicular bladder pressure. Anesthesia alters the urodynamics and in the conscious state this methodology requires that the dogs be tethered/restrained, which evokes stress and limits the period of continuous urodynamic assessment. A more physiological and chronic method of evaluating pharmacological responses on urodynamics is necessary. MATERIALS AND METHODS: Adult female beagle dogs were surgically instrumented with radiotelemetry transmitters enabling urodynamic/hemodynamic recordings. Telemetered urodynamics were compared to those measured in anesthetized dogs receiving bladder infusion of saline. The response to diuresis with furosemide (Intervet, Millsboro, Delaware) and the M3 selective antimuscarinic darifenacin (Matrix Laboratories, Hyderabad, India) were evaluated. RESULTS: Saline infused, anesthetized dogs demonstrated lower peak micturition pressure and higher threshold pressure than conscious, freely moving telemetered dogs. In telemetered dogs a single dose of furosemide increased voiding frequency and average urine volume per void. Darifenacin decreased peak voiding pressure without affecting voiding frequency. CONCLUSION: Telemetry provides the potential to significantly decrease animal use while enabling the continuous monitoring of urodynamics under more physiological conditions without tethering or artificial filling. In addition, this new model facilitates evaluation of the chronic efficacy of new urological therapies.


Asunto(s)
Telemetría , Urodinámica , Animales , Perros , Femenino
14.
Neurourol Urodyn ; 28(5): 442-6, 2009.
Artículo en Inglés | MEDLINE | ID: mdl-19030181

RESUMEN

AIMS: We tested cardiovascular and visceromotor reflex (VMR) responses to urinary bladder distension (UBD) in urethane anesthetized rats to see if it can replicate the response pattern and the inhibition of bladder nociceptive transmission by analgesics seen in isoflurane anesthetized animals. METHODS: Female Sprague-Dawley rats under 3% isoflurane anesthesia were acutely instrumented with jugular venous, carotid arterial, and bladder cannulas for drug administration, blood pressure (BP) measurement, and bladder distension, respectively. Needle electrodes were placed directly into the abdominal musculature to measure myoelectrical activity subsequent to phasic UBD (30 sec in 3 min intervals). A cardiovascular response (pressor) and a VMR response (a contraction of abdominal and hind limb musculature) to UBD were evaluated in urethane (1.2 g/kg, i.v.) or isoflurane (1%) anesthetized rats. RESULTS: Pressor and VMR responses to noxious UBD (60 mmHg) were generated under both anesthesics. The thresholds of stimulus response functions for both pressor and VMR responses were not affected by either anesthesics. However, the magnitude of the maximal pressor response was significantly reduced in urethane anesthesia. The analgesics, morphine, and mexiletine, significantly inhibited the VMR response to noxious UBD under both anesthetics, but the intensities of the inhibition from both analgesics under urethane anesthesia were much lower than under isoflurane anesthesia (ID50: 2.07 mg/kg vs. 0.88 mg/kg for morphine, >10 mg/kg vs. 0.47 mg/kg for mexiletine). CONCLUSIONS: The rat urinary bladder distension model in urethane anesthetized rats demonstrates a blunted maximal pressor response and a reduced inhibition of visceral nociceptive transmission by analgesics. Neurourol. Urodynam. 28:442-446, 2009. (c) 2008 Wiley-Liss, Inc.


Asunto(s)
Músculos Abdominales/inervación , Anestésicos Intravenosos/farmacología , Sistema Cardiovascular/inervación , Mecanotransducción Celular/efectos de los fármacos , Reflejo/efectos de los fármacos , Uretano/farmacología , Vejiga Urinaria/inervación , Analgésicos/farmacología , Anestésicos por Inhalación/farmacología , Animales , Presión Sanguínea/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Electromiografía , Femenino , Isoflurano/farmacología , Mexiletine/farmacología , Morfina/farmacología , Contracción Muscular/efectos de los fármacos , Presión , Ratas , Ratas Sprague-Dawley
15.
Cancer Res ; 68(18): 7475-83, 2008 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-18794135

RESUMEN

Androgens, through their actions on the androgen receptor (AR), are required for the development of the prostate and contribute to the pathologic growth dysregulation observed in prostate cancers. Consequently, androgen ablation has become an essential component of the pharmacotherapy of prostate cancer. In this study, we explored the utility of targeting processes downstream of AR as an alternate approach for therapy. Specifically, we show that the serum and glucocorticoid-regulated kinase 1 (SGK1) gene is an androgen-regulated target gene in cellular models of prostate cancer. Furthermore, functional serum- and glucocorticoid-regulated kinase 1 (SGK1) protein, as determined by the phosphorylation of its target Nedd4-2, was also increased with androgen treatment. Importantly, we determined that RNA interference-mediated knockdown of SGK1 expression attenuates the androgen-mediated growth of the prostate cancer cell line LNCaP. Given these findings, we explored the utility of SGK1 as a therapeutic target in prostate cancer by developing and evaluating a small-molecule inhibitor of this enzyme. From these studies emerged GSK650394, a competitive inhibitor that quantitatively blocks the effect of androgens on LNCaP cell growth. Thus, in addition to androgen ablation, inhibition of pathways downstream of AR is likely to have therapeutic utility in prostate cancer.


Asunto(s)
Proteínas Inmediatas-Precoces/antagonistas & inhibidores , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/enzimología , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Benzoatos/farmacología , Compuestos Bicíclicos Heterocíclicos con Puentes/farmacología , Procesos de Crecimiento Celular/fisiología , Línea Celular Tumoral , Células HeLa , Humanos , Proteínas Inmediatas-Precoces/biosíntesis , Proteínas Inmediatas-Precoces/genética , Proteínas Inmediatas-Precoces/metabolismo , Masculino , Metribolona/farmacología , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/patología , Proteínas Serina-Treonina Quinasas/biosíntesis , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , ARN Mensajero/biosíntesis , ARN Mensajero/genética , ARN Interferente Pequeño/genética , Receptores Androgénicos/metabolismo , Regulación hacia Arriba
16.
Am J Physiol Renal Physiol ; 295(4): F984-94, 2008 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-18632791

RESUMEN

Prostaglandin EP3 receptors in the central nervous system (CNS) may exert an excitatory effect on urinary bladder function via modulation of bladder afferent pathways. We have studied this action, using two EP3 antagonists, (2E)-3-{1-[(2,4-dichlorophenyl)methyl]-5-fluoro-3-methyl-1H-indol-7-yl}-N-[(4,5-dichloro-2-thienyl)sulfonyl]-2-propenamide (DG041) and (2E)-N-{[5-bromo-2-(methyloxy)phenyl] sulfonyl}-3-[2-(2-naphthalenylmethyl)phenyl]-2-propenamide (CM9). DG041 and CM9 were proven to be selective EP3 antagonists with radioligand binding and functional fluorescent imaging plate reader (FLIPR) assays. Their effects on volume-induced rhythmic bladder contraction and the visceromotor reflex (VMR) response to urinary bladder distension (UBD) were evaluated in female rats after intrathecal or intracerebroventricular administration. Both DG041 and CM9 showed a high affinity for EP3 receptors at subnanomolar concentrations without significant selectivity for any splice variants. At the human EP3C receptor, both inhibited calcium influx produced by the nonselective agonist PGE2. After intrathecal or intracerebroventricular administration both CM9 and DG041 dose-dependently reduced the frequency, but not the amplitude, of the bladder rhythmic contraction. With intrathecal administration DG041 and CM9 produced a long-lasting and robust inhibition on the VMR response to UBD, whereas with intracerebroventricular injection both compounds elicited only a transient reduction of the VMR response to bladder distension. These data support the concept that EP3 receptors are involved in bladder micturition at supraspinal and spinal centers and in bladder nociception at the spinal cord. A centrally acting EP3 receptor antagonist may be useful in the control of detrusor overactivity and/or pain associated with bladder disorders.


Asunto(s)
Sistema Nervioso Central/fisiología , Receptores de Prostaglandina E/metabolismo , Reflejo/fisiología , Vejiga Urinaria/inervación , Vejiga Urinaria/fisiología , Acrilamidas/química , Acrilamidas/farmacología , Animales , Células CHO , Línea Celular Tumoral , Sistema Nervioso Central/efectos de los fármacos , Cricetinae , Cricetulus , Dinoprostona/metabolismo , Femenino , Humanos , Inyecciones Intraventriculares , Inyecciones Espinales , Riñón/citología , Contracción Muscular/efectos de los fármacos , Contracción Muscular/fisiología , Nociceptores/fisiología , Osteosarcoma , Ratas , Ratas Sprague-Dawley , Receptores de Prostaglandina E/antagonistas & inhibidores , Receptores de Prostaglandina E/genética , Subtipo EP3 de Receptores de Prostaglandina E , Reflejo/efectos de los fármacos , Sulfonas/química , Sulfonas/farmacología , Transfección , Tritio , Micción/fisiología
17.
Am J Physiol Renal Physiol ; 295(2): F585-94, 2008 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-18562635

RESUMEN

The excitatory roles of EP3 receptors at the peripheral afferent nerve innervating the rat urinary bladder have been evaluated by using the selective EP3 antagonist (2E)-3-[1-[(2,4-dichlorophenyl)methyl]-5-fluoro-3-methyl-1H-indol-7-yl]-N-[(4,5-dichloro-2-thienyl)sulfonyl]-2-propenamide (DG-041). The bladder rhythmic contraction model and a bladder pain model measuring the visceromotor reflex (VMR) to urinary bladder distension (UBD) have been used to evaluate DG-041 in female rats. In addition, male rats [spontaneously hypertensive rat (SHR), Wistar-Kyoto (WKY), and Sprague-Dawley (SD)] were anesthetized with pentobarbital sodium, and primary afferent fibers in the L6 dorsal root were isolated for recording the inhibitory response to UBD following intravenous injection of DG-041. Intravenous injection of DG-041 (10 mg/kg), a peripherally restricted EP3 receptor antagonist, significantly reduced the frequency of bladder rhythmic contraction and inhibited the VMR response to bladder distension. The magnitude of reduction of the VMR response was not different in the different strains of rats (SD, SHR, and WKY). Furthermore, quantitative characterization of the mechanosensitive properties of bladder afferent nerves in SHR, WKY, and SD rats did not show the SHR to be supersensitive to bladder distension. DG-041 selectively attenuated responses of mechanosensitive afferent nerves to UBD, with strong suppression on the slow-conducting, high-threshold afferent fibers, with equivalent activity in the three strains. We conclude that sensitization of afferent nerve activity was not one of the mechanisms of bladder hypersensitivity in SHR. EP3 receptors are involved in the regulation of bladder micturition and bladder nociception at the peripheral level.


Asunto(s)
Neuronas Aferentes/fisiología , Receptores de Prostaglandina E/fisiología , Vejiga Urinaria/inervación , Vejiga Urinaria/fisiología , Acrilamidas/farmacología , Animales , Ciclofilinas/metabolismo , Modelos Animales de Enfermedad , Femenino , Gliceraldehído-3-Fosfato Deshidrogenasas/metabolismo , Hipertensión/fisiopatología , Masculino , Contracción Muscular/efectos de los fármacos , Músculo Liso/inervación , Músculo Liso/metabolismo , Músculo Liso/fisiología , Neuronas Aferentes/efectos de los fármacos , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Ratas Sprague-Dawley , Receptores de Prostaglandina E/antagonistas & inhibidores , Receptores de Prostaglandina E/efectos de los fármacos , Subtipo EP3 de Receptores de Prostaglandina E , Sulfonas/farmacología , Vejiga Urinaria/metabolismo
18.
Am J Physiol Renal Physiol ; 295(3): F803-10, 2008 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-18562636

RESUMEN

The activation of the TRPM8 channel, a member of the large class of TRP ion channels, has been reported to be involved in overactive bladder and painful bladder syndrome, although an endogenous activator has not been identified. In this study, N-(3-aminopropyl)-2-{[(3-methylphenyl) methyl]oxy}-N-(2-thienylmethyl)benzamide hydrochloride salt (AMTB) was evaluated as a TRPM8 channel blocker and used as a tool to evaluate the effects of this class of ion channel blocker on volume-induced bladder contraction and nociceptive reflex responses to noxious bladder distension in the rat. AMTB inhibits icilin-induced TRPM8 channel activation as measured in a Ca(2+) influx assay, with a pIC(50) of 6.23. In the anesthetized rat, intravenous administration of AMTB (3 mg/kg) decreased the frequency of volume-induced bladder contractions, without reducing the amplitude of contraction. The nociceptive response was measured by analyzing both visceromotor reflex (VMR) and cardiovascular (pressor) responses to urinary bladder distension (UBD) under 1% isoflurane. AMTB (10 mg/kg) significantly attenuated reflex responses to noxious UBD to 5.42 and 56.51% of the maximal VMR response and pressor response, respectively. The ID50 value on VMR response was 2.42 +/- 0.46 mg/kg. These results demonstrate that TRPM8 channel blocker can act on the bladder afferent pathway to attenuate the bladder micturition reflex and nociceptive reflex responses in the rat. Targeting TRPM8 channel may provide a new therapeutic opportunity for overactive bladder and painful bladder syndrome.


Asunto(s)
Benzamidas/farmacología , Contracción Muscular/efectos de los fármacos , Presorreceptores/efectos de los fármacos , Canales Catiónicos TRPM/metabolismo , Tiofenos/farmacología , Vejiga Urinaria Hiperactiva/metabolismo , Vías Aferentes/efectos de los fármacos , Animales , Benzamidas/farmacocinética , Femenino , Expresión Génica , Reacción en Cadena de la Polimerasa , Ratas , Ratas Sprague-Dawley , Reflejo/efectos de los fármacos , Canales Catiónicos TRPM/antagonistas & inhibidores , Tiofenos/farmacocinética
19.
Am J Physiol Renal Physiol ; 295(2): F507-14, 2008 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-18508878

RESUMEN

Nonsteroidal anti-inflammatory cyclooxygenase inhibitors that function to reduce prostaglandin E2 (PGE2) production have been widely reported as effective agents in models of urinary bladder overactivity. We therefore investigated a potential role for the PGE2 receptor, EP3, in urinary bladder function by performing conscious, freely moving cystometry on EP3 receptor knockout (KO) mice. EP3 KO mice demonstrated an enhanced bladder capacity compared with wild-type (WT) mice ( approximately 185% of WT) under control conditions, based on larger voided and infused bladder volumes. Infusion of the EP3 receptor agonist GR63799X into the bladder of WT mice reduced the bladder capacity. This was ineffective in EP3 KO mice that demonstrated a time-dependent increase in bladder capacity with GR63799X, an effect similar to that observed with vehicle in both genotypes. In addition, infusion of PGE2 into WT mice induced bladder overactivity, an effect that was significantly blunted in the EP3 KO mice. The data reported here provide the first evidence supporting a functional role for EP3 receptors in normal urinary bladder function and implicate EP3 as a contributor to bladder overactivity during pathological conditions of enhanced PGE2 production, as reported previously in overactive bladder patients.


Asunto(s)
Dinoprostona/metabolismo , Receptores de Prostaglandina E/metabolismo , Vejiga Urinaria Hiperactiva/metabolismo , Vejiga Urinaria/patología , Vejiga Urinaria/fisiopatología , Ácido Acético , Animales , Modelos Animales de Enfermedad , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Prostaglandinas E Sintéticas/farmacología , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Receptores de Prostaglandina E/efectos de los fármacos , Receptores de Prostaglandina E/genética , Subtipo EP3 de Receptores de Prostaglandina E , Vejiga Urinaria Hiperactiva/inducido químicamente
20.
J Pharmacol Exp Ther ; 326(2): 432-42, 2008 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-18499743

RESUMEN

The transient receptor potential (TRP) vanilloid 4 (TRPV4) member of the TRP superfamily has recently been implicated in numerous physiological processes. In this study, we describe a small molecule TRPV4 channel activator, (N-((1S)-1-{[4-((2S)-2-{[(2,4-dichlorophenyl)sulfonyl]amino}-3-hydroxypropanoyl)-1-piperazinyl]carbonyl}-3-methylbutyl)-1-benzothiophene-2-carboxamide (GSK1016790A), which we have used as a valuable tool in investigating the role of TRPV4 in the urinary bladder. GSK1016790A elicited Ca2+ influx in mouse and human TRPV4-expressing human embryonic kidney (HEK) cells (EC50 values of 18 and 2.1 nM, respectively), and it evoked a dose-dependent activation of TRPV4 whole-cell currents at concentrations above 1 nM. In contrast, the TRPV4 activator 4alpha-phorbol 12,13-didecanoate (4alpha-PDD) was 300-fold less potent than GSK1016790A in activating TRPV4 currents. TRPV4 mRNA was detected in urinary bladder smooth muscle (UBSM) and urothelium of TRPV4+/+ mouse bladders. Western blotting and immunohistochemistry demonstrated protein expression in both the UBSM and urothelium that was absent in TRPV4-/- bladders. TRPV4 activation with GSK1016790A contracted TRPV4+/+ mouse bladders in vitro, both in the presence and absence of the urothelium, an effect that was undetected in TRPV4-/- bladders. Consistent with the effects on TRPV4 HEK whole-cell currents, 4alpha-PDD demonstrated a weak ability to contract bladder strips compared with GSK1016790A. In vivo, urodynamics in TRPV4+/+ and TRPV4-/- mice revealed an enhanced bladder capacity in the TRPV4-/- mice. Infusion of GSK1016790A into the bladders of TRPV4+/+ mice induced bladder overactivity with no effect in TRPV4-/- mice. Overall TRPV4 plays an important role in urinary bladder function that includes an ability to contract the bladder as a result of the expression of TRPV4 in the UBSM.


Asunto(s)
Leucina/análogos & derivados , Contracción Muscular/efectos de los fármacos , Sulfonamidas/farmacología , Canales Catiónicos TRPV/agonistas , Vejiga Urinaria/efectos de los fármacos , Urodinámica/efectos de los fármacos , Urotelio/efectos de los fármacos , Animales , Peso Corporal/efectos de los fármacos , Femenino , Leucina/farmacología , Masculino , Ratones , Ratones Noqueados , Estructura Molecular , Músculo Liso/efectos de los fármacos , Músculo Liso/metabolismo , Forboles/farmacología , Canales Catiónicos TRPV/genética , Canales Catiónicos TRPV/fisiología , Vejiga Urinaria/metabolismo , Urotelio/metabolismo
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA