RESUMEN
BACKGROUND: E2027 is a novel, highly selective and potent inhibitor of phosphodiesterase9 (PDE9) being evaluated as a treatment for dementia with Lewy bodies. METHODS: Phase 1, randomized, double-blind, single ascending dose (SAD, n=96) and multiple ascending dose (MAD, n=68) studies evaluated E2027 doses (5 to 1200 mg) in healthy subjects. The impact of age, race (Japanese/non-Japanese), and food on pharmacokinetics (PK)/pharmacodynamics were evaluated. Serial cerebrospinal fluid (CSF) samples were collected to assess the target engagement. RESULTS: E2027 PK profiles were biphasic (elimination half-life: ~30 hours. Approximately 3-fold accumulation was observed following multiple once-daily dosing. E2027 single doses of 50 to 400 mg resulted in mean maximum increases in CSF cyclic guanosine monophosphate ranging from 293% to 461% within 5.37 to 12.9 hours after dose administration to assess target engagement. Dose-response modelling of steady-state predose CSF cyclic guanosine monophosphate concentrations showed ≥200% increase from baseline is maintained with doses of ≥50 mg QD. The most common adverse events with E2027 were post-LP syndrome and back pain. PK profiles were similar between Japanese and non-Japanese. Higher exposure observed in fed versus fasted state was not considered clinically significant. PK exposure was higher in elderly subjects. CONCLUSIONS: S.E2027 was well-tolerated following single and multiple administration. E2027 achieved maximal and sustained target engagement at 50 mg QD, the dose selected for the phase 2 clinical trial.
Asunto(s)
Enfermedad por Cuerpos de Lewy , Inhibidores de Fosfodiesterasa , Anciano , Área Bajo la Curva , Ensayos Clínicos Fase II como Asunto , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Guanosina Monofosfato , Voluntarios Sanos , Humanos , Enfermedad por Cuerpos de Lewy/tratamiento farmacológico , Inhibidores de Fosfodiesterasa/uso terapéutico , Hidrolasas Diéster FosfóricasRESUMEN
N-Cyclopropylmethyl-7-(2,6-dimethoxy-4-methoxymethylphenyl)-2-ethyl-N-(tetrahydro-2H-pyran-4-ylmethyl)pyrazolo[1,5-a]pyridin-3-amine tosylate (E2508) is a newly discovered selective corticotropin-releasing factor 1 receptor antagonist. Here, we investigated the effects of E2508 on wrap restraint stress-induced defecation and visceral pain in rats. Oral pretreatment with E2508 dose-dependently decreased stool weights after 20min wrap restraint stress and significant effects were observed at doses of 30 and 100mg/kg. However, E2508 did not affect basal defecation at doses up to 100mg/kg. In contrast, alosetron, a 5-HT3 receptor antagonist, decreased both wrap restraint stress-induced and basal stool output at a dose of 0.1mg/kg. In a rat visceral pain model, subcutaneous injections of both E2508 (0.01 and 0.1mg/kg) and alosetron (0.001 and 0.01mg/kg) significantly decreased the number of abdominal muscle contractions induced by colonic distention, suggesting these drugs reduced visceral pain. Together, these results demonstrate E2508 has the potential to be an effective therapy for the treatment of irritable bowel syndrome with a lower risk of adverse events such as constipation compared with the current clinically used 5-HT3 receptor antagonist.
Asunto(s)
Defecación/efectos de los fármacos , Síndrome del Colon Irritable/tratamiento farmacológico , Pirazoles/farmacología , Piridinas/farmacología , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Antagonistas del Receptor de Serotonina 5-HT3/farmacología , Estrés Psicológico/complicaciones , Dolor Visceral/tratamiento farmacológico , Animales , Carbolinas/administración & dosificación , Carbolinas/farmacología , Modelos Animales de Enfermedad , Masculino , Pirazoles/administración & dosificación , Piridinas/administración & dosificación , Ratas , Ratas Endogámicas F344 , Antagonistas del Receptor de Serotonina 5-HT3/administración & dosificación , Dolor Visceral/etiologíaRESUMEN
Corticotropin-releasing factor (CRF) is a hormone secreted by the hypothalamus in response to stress, and CRF antagonists may be effective for the treatment of stress-related disorders including major depressive and anxiety disorders. Here, we investigated the in vivo pharmacological profile of N-cyclopropylmethyl-7-(2,6-dimethoxy-4-methoxymethylphenyl)-2-ethyl-N-(tetrahydro-2H-pyran-4-ylmethyl)pyrazolo[1,5-a]pyridin-3-amine tosylate (E2508), a recently synthesized, orally active CRF1 receptor antagonist. Oral administration of a single dose of E2508 (3 or 10mg/kg), but not fluoxetine (30mg/kg), a selective serotonin reuptake inhibitor (SSRI), significantly shortened immobility time in rats in the forced swim test. E2508 (10, 30, or 100mg/kg) also showed an antidepressant-like effect in the forced swim test in mice, with no sedative or muscle relaxant effects for doses up to 100mg/kg. Moreover, E2508 (5 or 20mg/kg) significantly reduced anxiety-like behavior in the rat defensive burying test. Diazepam, a benzodiazepine anxiolytic agent, also showed an anxiolytic effect in the defensive burying test at the same dose that induced a muscle relaxant effect in mice. Administration of E2508 (30mg/kg) for 14 consecutive days did not affect sexual behavior. By contrast, fluoxetine (30mg/kg) administration for ≥7 consecutive days decreased sexual behavior. These results indicate that E2508 has both potent antidepressant-like and anxiolytic-like effects in rodent models, and is well tolerated compared with a commonly prescribed therapeutic SSRI or benzodiazepine.
Asunto(s)
Ansiolíticos/administración & dosificación , Antidepresivos/administración & dosificación , Trastornos de Ansiedad/prevención & control , Trastorno Depresivo Mayor/prevención & control , Pirazoles/administración & dosificación , Piridinas/administración & dosificación , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Acetilcolina/sangre , Administración Oral , Animales , Trastornos de Ansiedad/metabolismo , AMP Cíclico/metabolismo , Trastorno Depresivo Mayor/metabolismo , Diazepam/administración & dosificación , Modelos Animales de Enfermedad , Femenino , Fluoxetina/administración & dosificación , Células HEK293 , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Fuerza Muscular/efectos de los fármacos , Pirimidinas/administración & dosificación , Ratas , Ratas Endogámicas F344 , Ratas Sprague-Dawley , Ratas Wistar , Receptores de Hormona Liberadora de Corticotropina/metabolismo , Inhibidores Selectivos de la Recaptación de Serotonina/administración & dosificación , Conducta Sexual Animal/efectos de los fármacosRESUMEN
We previously reported a series of 8-methyl-2-aryl-5-alkylaminoquinolines as a novel class of corticotropin-releasing factor-1 (CRF(1)) receptor antagonists. A critical issue encountered for this series of compounds was low aqueous solubility at physiological pH (pH 7.4). To address this issue, derivatization at key sites (R(2), R(3), R(5), R(5'), and R(8)) was performed and the relationships between structure and solubility were examined. As a result, it was revealed that introduction of a methoxy substituent at the C(8) position had a positive impact on the solubility of the derivatives. Consequently, through in vivo and in vitro biological studies, compound 21d was identified as a potent, orally active CRF(1) receptor antagonist with improved physicochemical properties.
Asunto(s)
Aminoquinolinas/química , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Administración Oral , Aminoquinolinas/síntesis química , Aminoquinolinas/farmacocinética , Animales , Conducta Animal/efectos de los fármacos , Diseño de Fármacos , Semivida , Concentración de Iones de Hidrógeno , Ratones , Ratones Endogámicos BALB C , Ratas , Receptores de Hormona Liberadora de Corticotropina/metabolismo , Solubilidad , Relación Estructura-ActividadRESUMEN
This paper describes the design, synthesis, and structure-activity relationships of a novel series of 7-dialkylamino-3-phenyl-6-methoxy pyrazolo[5,1-b]thiazole derivatives for use as selective antagonists of the corticotropin-releasing factor 1 (CRF(1)) receptor. The most promising compound, N-butyl-3-[4-(ethoxymethyl)-2,6-dimethoxyphenyl]-6-methoxy-N-(tetrahydro-2H-pyran-4-yl)pyrazolo[5,1-b][1,3]thiazole-7-amine (6t), showed high affinity (IC(50) = 70 nM) and functional antagonism (IC(50) = 7.1 nM) for the human CRF(1) receptor as well as dose-dependent inhibition of the CRF-induced increase in the plasma adrenocorticotropic hormone (ACTH) concentration at a dose of 30 mg/kg (po). Further, in the light/dark test in mice, the compound 6t showed anxiolytic activity at a dose of 30 mg/kg (po).
Asunto(s)
Ansiolíticos/síntesis química , Pirazoles/síntesis química , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Tiazoles/síntesis química , Administración Oral , Hormona Adrenocorticotrópica/sangre , Animales , Ansiolíticos/química , Ansiolíticos/farmacología , Línea Celular Tumoral , Hormona Liberadora de Corticotropina/farmacología , AMP Cíclico/metabolismo , Defecación/efectos de los fármacos , Diseño de Fármacos , Células HEK293 , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Modelos Moleculares , Pirazoles/química , Pirazoles/farmacología , Ensayo de Unión Radioligante , Ratas , Ratas Endogámicas F344 , Ratas Sprague-Dawley , Relación Estructura-Actividad , Tiazoles/química , Tiazoles/farmacologíaRESUMEN
We designed and synthesized a series of 2-Ar-8-methyl-5-alkylaminolquinolines as potent corticotropin-releasing factor 1 (CRF(1)) receptor antagonists. The structure-activity relationships of substituents at each position (R(3), R(5), R(5'), and R(8)) was investigated. By derivatization, three compounds (6, 14b, and 14c) were identified as orally active CRF(1) receptor antagonists.
Asunto(s)
Quinolinas/química , Quinolinas/farmacología , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Administración Oral , Animales , Depresión/tratamiento farmacológico , Diseño de Fármacos , Células HEK293 , Humanos , Modelos Moleculares , Quinolinas/administración & dosificación , Quinolinas/síntesis química , Ratas , Receptores de Hormona Liberadora de Corticotropina/metabolismo , Relación Estructura-ActividadRESUMEN
A series of 5-alkylaminolquinolines was designed and synthesized as potential novel CRF(1) receptor antagonists. The structure-activity relationships (SARs) of the substituents on each position (R(2), R(3), R(5) and R(5')) were investigated.
Asunto(s)
Quinolinas/síntesis química , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Alquilación , Aminación , Diseño de Fármacos , Células HEK293 , Humanos , Modelos Moleculares , Estructura Molecular , Quinolinas/farmacología , Relación Estructura-ActividadRESUMEN
Design, synthesis, and structure-activity relationships of a series of 3-dialkylamino-7-phenyl pyrazolo[1,5-a]pyridines (I) as selective antagonists of the corticotropin-releasing factor 1 (CRF(1)) receptor are described. The most prominent compound to emerge from this work, 46 (E2508), exhibits potent in vitro activity, excellent drug-like properties, and robust oral efficacy in animal models of stress-related disorders. It has advanced into clinical trials.
Asunto(s)
Antidepresivos/síntesis química , Pirazoles/síntesis química , Piridinas/síntesis química , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Administración Oral , Animales , Antidepresivos/farmacocinética , Antidepresivos/farmacología , Disponibilidad Biológica , AMP Cíclico/biosíntesis , Células HEK293 , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Microsomas Hepáticos/metabolismo , Pirazoles/farmacocinética , Pirazoles/farmacología , Piridinas/farmacocinética , Piridinas/farmacología , Relación Estructura-ActividadRESUMEN
BACKGROUND: ADAM22 is a member of the ADAM gene family, but the fact that it is expressed only in the nervous systems makes it unique. ADAM22's sequence similarity to other ADAMs suggests it to be an integrin binder and thus to have a role in cell-cell or cell-matrix interactions. To elucidate the physiological functions of ADAM22, we employed gene targeting to generate ADAM22 knockout mice. RESULTS: ADAM22-deficient mice were produced in a good accordance with the Mendelian ratio and appeared normal at birth. After one week, severe ataxia was observed, and all homozygotes died before weaning, probably due to convulsions. No major histological abnormalities were detected in the cerebral cortex or cerebellum of the homozygous mutants; however, marked hypomyelination of the peripheral nerves was observed. CONCLUSION: The results of our study demonstrate that ADAM22 is closely involved in the correct functioning of the nervous system. Further analysis of ADAM22 will provide clues to understanding the mechanisms of human diseases such as epileptic seizures and peripheral neuropathy.
Asunto(s)
Proteínas ADAM/deficiencia , Proteínas ADAM/fisiología , Ataxia/metabolismo , Fibras Nerviosas Mielínicas/metabolismo , Proteínas del Tejido Nervioso/deficiencia , Proteínas del Tejido Nervioso/fisiología , Enfermedades del Sistema Nervioso Periférico/metabolismo , Proteínas ADAM/genética , Animales , Ataxia/genética , Ataxia/patología , Células Cultivadas , Femenino , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Fibras Nerviosas Mielínicas/patología , Proteínas del Tejido Nervioso/genética , Nervios Periféricos/metabolismo , Nervios Periféricos/patología , Enfermedades del Sistema Nervioso Periférico/genética , Enfermedades del Sistema Nervioso Periférico/patologíaRESUMEN
The voltage-dependent N-type Ca2+ channel is localized in the plasma membrane of insulin-releasing beta-cells and glucagon-releasing alpha-cells in the islets of Langerhans in the pancreas. To examine the contribution of N-type Ca2+ channel to glucose homeostasis, we performed glucose tolerance and insulin tolerance tests with N-type Ca2+ channel alpha(1B)-subunit-deficient mice on a normal or high-fat diet. The fasting glucose level in homozygous mice on the normal diet was significantly lower than those in wild and heterozygous mice. In glucose tolerance tests, the homozygous mice showed a higher glucose clearance rate and a similar pattern of insulin levels to those of wild and heterozygous mice. In insulin tolerance tests, glucose clearance rates showed no significant difference among wild, heterozygous and homozygous mice. In animals on the high-fat diet, food consumption was the same among wild, heterozygous and homozygous mice, but body weight gain was reduced in homozygous mice. After 8 weeks of the high-fat diet, homozygous mice showed lower fasting glucose levels and exhibited higher glucose clearance and lower insulin levels than wild or heterozygous mice in glucose tolerance tests. Glucose clearance rates showed no significant difference among wild, heterozygous and homozygous mice in insulin tolerance tests. After 10 weeks of the high-fat diet, the alpha(1B)-deficient homozygous mice showed lower lipid deposition in liver and lower plasma glucagon, leptin and triglyceride levels than wild or heterozygous mice. These results suggest that N-type Ca2+ channels play a role in insulin and glucagon release, and that N-type Ca2+ channel alpha(1B)-subunit deficient mice show improved glucose tolerance without any change in insulin sensitivity. Thus, N-type Ca2+ channel blockers might be candidate anti-diabetic/anti-obesity agents.
Asunto(s)
Canales de Calcio Tipo N/deficiencia , Glucosa/administración & dosificación , Subunidades de Proteína/metabolismo , Animales , Glucemia/análisis , Glucemia/metabolismo , Peso Corporal/efectos de los fármacos , Canales de Calcio Tipo N/genética , Canales de Calcio Tipo N/metabolismo , Cruzamientos Genéticos , Grasas de la Dieta/administración & dosificación , Grasas de la Dieta/farmacología , Ingestión de Alimentos/efectos de los fármacos , Ayuno , Glucagón/metabolismo , Glucosa/metabolismo , Glucosa/farmacología , Prueba de Tolerancia a la Glucosa , Heterocigoto , Homeostasis , Homocigoto , Insulina/sangre , Insulina/metabolismo , Resistencia a la Insulina , Secreción de Insulina , Islotes Pancreáticos/efectos de los fármacos , Islotes Pancreáticos/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos CBA , Ratones Noqueados , Obesidad/genética , Obesidad/metabolismo , Obesidad/patología , Reacción en Cadena de la PolimerasaRESUMEN
The Ca(2+) channel alpha(1B) subunit is a pore-forming component capable of generating N-type Ca(2+) channel activity. Although the N-type Ca(2+) channel plays a role in a variety of neuronal functions, alpha(1B)-deficient mice show normal behavior, presumably owing to compensation by the other Ca(2+) channels. In this study, we examined the mRNA expression of the P/Q-type Ca(2+) channel alpha(1A) subunit in cerebellum of alpha(1B)-deficient mice. The alpha(1A) subunit mRNA in homozygous alpha(1B)-deficient mice was expressed at a significantly higher level than in wild or heterozygous mice. To examine whether the increased expression is induced by a cis-regulatory element within the 5'-upstream region of the alpha(1A) subunit gene, we examined lacZ expression in alpha(1B)-deficient x alpha(1A)3.0-lacZ mice (carrying a 3.0-kb 5'-upstream fragment of the alpha(1A) subunit gene fused to Escherichia coli lacZ reporter gene), which express lacZ in granule but not in Purkinje cells, and in alpha(1B)-deficient x alpha(1A)6.3-lacZ mice (carrying a 6.3-kb 5'-upstream region fused to lacZ gene), which express lacZ in Purkinje but not in granule cells. The levels of lacZ expression in homozygous alpha(1B)-deficient x alpha(1A)3.0-lacZ mice were significantly higher than in wild or heterozygous mice, but no difference in lacZ expression level was found among wild, heterozygous and homozygous alpha(1B)-deficient x alpha(1A)6.3-lacZ mice. Therefore, a possible explanation of the normal behavior of alpha(1B)-deficient mice is that compensation by alpha(1A) subunit gene occurs and that the 3.0-kb 5'-upstream region of alpha(1A) subunit gene contains an enhancer cis-element(s) for compensation in cerebellar granule cells.
Asunto(s)
Canales de Calcio Tipo N/deficiencia , Canales de Calcio Tipo P/metabolismo , Canales de Calcio Tipo Q/metabolismo , Cerebelo/metabolismo , Subunidades de Proteína/metabolismo , Animales , Northern Blotting/métodos , Canales de Calcio Tipo N/genética , Canales de Calcio Tipo N/metabolismo , Cerebelo/citología , Operón Lac/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neuronas/metabolismo , ARN Mensajero/metabolismo , beta-Galactosidasa/metabolismoRESUMEN
N-type and P/Q-type Ca2+ channels play an important role in the processing of olfactory information. However, N-type Ca2+ channel alpha1B-deficient mice show normal behavior, presumably owing to compensation by other Ca2+ channels. P/Q-type Ca2+ channel alpha1A mRNA was expressed at a higher level in olfactory bulb of homozygous alpha1B-deficient mice than wild-type or heterozygous mice. LacZ expression in olfactory mitral cells of homozygous alpha1B-deficient x alpha1A1.5-lacZ mice, carrying a 1.5-kb 5'-upstream fragment of the alpha1A gene fused to the lacZ reporter gene, was increased compared to that in wild-type or heterozygous mice. Therefore, a possible explanation for the normal behavior of alpha1B-deficient mice is compensation by the alpha1A gene and that the 1.5-kb 5'-upstream region of this gene contains an enhancer cis-element for compensation in olfactory mitral cells.
Asunto(s)
Canales de Calcio/biosíntesis , Bulbo Olfatorio/citología , Bulbo Olfatorio/metabolismo , ARN Mensajero/biosíntesis , Animales , Canales de Calcio/genética , Canales de Calcio/fisiología , Canales de Calcio Tipo N/biosíntesis , Canales de Calcio Tipo N/deficiencia , Canales de Calcio Tipo N/genética , Canales de Calcio Tipo P , Canales de Calcio Tipo Q , Regulación de la Expresión Génica/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos CBA , Ratones Noqueados , ARN Mensajero/genética , beta-Galactosidasa/biosíntesisRESUMEN
Although the N-type Ca2+ channel plays a role in a variety of neuronal functions, N-type Ca2+ channel alpha1B-deficient mice exhibit normal life span without apparent behavioral or histologic abnormalities. To examine whether the reason for their normal behavior is compensation by other Cav2 channel alpha1 or beta subunit genes and to analyze whether genetic background influences the subunit expression pattern, we studied the alpha1A, alpha1E, beta1b, beta2, beta3 and beta4 subunit mRNA levels in cerebellum of alpha1B-deficient mice with CBA x C57BL/6 or CBA/JN background. In cerebellum of the mice with a CBA x C57BL/6 background, alpha1A mRNA was expressed at a higher level than that in wild-type or heterozygous mice, but difference in the expression levels of alpha1E, beta1b, beta2, beta3 and beta4 subunits was not found among wild-type, heterozygous, and homozygous mice. In cerebellum of alpha1B-deficient mice with CBA/JN background, beta4 mRNA was expressed at a higher level than that in wild-type or heterozygous mice, but alpha1A, alpha1E, beta1b, alpha2, beta3 and transcripts were expressed at similar levels in all genotypes. Therefore, a possible explanation of the normal behavior of alpha1B-deficient mice is that Cav2 channel family members compensate for the deficiency, and that the change of functional subunit expression pattern for compensation differs in animals with different genetic backgrounds.
Asunto(s)
Canales de Calcio/genética , Cerebelo/metabolismo , Animales , Canales de Calcio/química , Canales de Calcio/deficiencia , Femenino , Expresión Génica , Masculino , Ratones , Ratones Endogámicos CBA , Ratones Endogámicos ICR , Ratones Noqueados , Subunidades de Proteína , ARN Mensajero/genética , ARN Mensajero/metabolismo , Especificidad de la EspecieRESUMEN
Differential properties of voltage-dependent Ca2+ channels have been primarily ascribed to the alpha1 subunit, of which 10 different subtypes are currently known. For example, channels that conduct the N-type Ca2+ current possess the alpha1B subunit (Cav2.2), which has been localized, inter alia, to the piriform cortex, hippocampus, hypothalamus, locus coeruleus, dorsal raphe, thalamic nuclei, and granular layer of the cortex. Some of these regions have been previously implicated in metabolic and vigilance state control, and selective block of the N-type Ca2+ channel causes circadian rhythm disruption. In this study of Cav2.2-/- knock-out mice, we examined potential differences in feeding behavior, spontaneous locomotion, and the sleep-wake cycle. Cav2.2-/- mice did not display an overt metabolic phenotype but were hyperactive, demonstrating a 20% increase in activity under novel conditions and a 95% increase in activity under habituated conditions during the dark phase, compared with wild-type littermates. Cav2.2-/- mice also displayed vigilance state differences during the light phase, including increased consolidation of rapid-eye movement (REM) sleep and increased intervals between non-REM (NREM) and wakefulness episodes. EEG spectral power was increased during wakefulness and REM sleep and was decreased during NREM sleep in Cav2.2-/- mice. These results indicate a role of the N-type Ca2+ channel in activity and vigilance state control, which we interpret in terms of effects on neurotransmitter release.
Asunto(s)
Nivel de Alerta/genética , Canales de Calcio Tipo N/deficiencia , Canales de Calcio Tipo N/genética , Hipercinesia/genética , Animales , Electroencefalografía , Electromiografía , Conducta Alimentaria/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Actividad Motora/genética , Actividad Motora/fisiología , Subunidades de Proteína/deficiencia , Subunidades de Proteína/genética , Fases del Sueño/genética , Fases del Sueño/fisiología , Sueño REM/genética , Sueño REM/fisiología , Vigilia/genética , Vigilia/fisiologíaRESUMEN
N-type voltage-dependent Ca(2+) channels (VDCCs), predominantly localized in the nervous system, have been proposed to play vital roles in a variety of neuronal functions such as neurotransmitter release at sympathetic nerve terminals. To directly approach the elucidation of the physiological significance of N-type VDCCs in the autonomic nervous system, alpha(1B) subunit (Ca(V) 2.2)-deficient mice were generated, in which peripheral neurons showed a complete and selective elimination of N-type channel currents sensitive to omega-conotoxin GVIA (the peptide toxin from the fish-hunting cone snail Conus geographus), without a significant effect on the activity of other VDCC types. In isolated left atria prepared from N-type-deficient mice, the positive inotropic response mediated by the sympathetic nervous system was dramatically decreased, whereas the negative inotropic response mediated by parasympathetic neurons was nearly intact compared with those of normal mice. The baroreflex response to bilateral carotid occlusion was markedly reduced in the mutant mice. Interestingly, the mutant mice showed sustained elevation of heart rate and blood pressure. These results provide direct in vivo evidence for an essential role of N-type VDCCs in maintaining the normal function of the sympathetic nervous system in circulatory regulation, demonstrating a potential of N-type VDCC-deficient mice as a useful model for studying disorders attributable to sympathetic nerve dysfunction.