RESUMEN
In this Phase 2b study, 331 low-to-moderate risk de novo kidney transplant patients (approximately 60% deceased donors) were randomized to a more intensive (MI) or less intensive (LI) regimen of tofacitinib (CP-690, 550), an oral Janus kinase inhibitor or cyclosporine (CsA). All patients received basiliximab induction, mycophenolic acid and corticosteroids. Primary endpoints were: incidence of biopsy-proven acute rejection (BPAR) with a serum creatinine increase of ≥0.3 mg/dL and ≥20% (clinical BPAR) at Month 6 and measured GFR at Month 12. Similar 6-month incidences of clinical BPAR (11%, 7% and 9%) were observed for MI, LI and CsA. Measured GFRs were higher (p < 0.01) at Month 12 for MI and LI versus CsA (65 mL/min, 65 mL/min vs. 54 mL/min). Fewer (p < 0.05) patients in MI or LI developed chronic allograft nephropathy at Month 12 compared with CsA (25%, 24% vs. 48%). Serious infections developed in 45%, 37% and 25% of patients in MI, LI and CsA, respectively. Anemia, neutropenia and posttransplant lymphoproliferative disorder occurred more frequently in MI and LI compared with CsA. Tofacitinib was equivalent to CsA in preventing acute rejection, was associated with improved renal function and less chronic allograft histological injury, but had side-effects at the doses evaluated.
Asunto(s)
Inmunosupresores/uso terapéutico , Janus Quinasa 3/antagonistas & inhibidores , Trasplante de Riñón , Inhibidores de Proteínas Quinasas/uso terapéutico , Pirimidinas/uso terapéutico , Pirroles/uso terapéutico , Adulto , Femenino , Humanos , Inmunosupresores/efectos adversos , Inmunosupresores/farmacocinética , Masculino , Persona de Mediana Edad , Piperidinas , Inhibidores de Proteínas Quinasas/efectos adversos , Inhibidores de Proteínas Quinasas/farmacocinética , Pirimidinas/efectos adversos , Pirimidinas/farmacocinética , Pirroles/efectos adversos , Pirroles/farmacocinéticaRESUMEN
Inhibition of alpha4beta1/vascular cell adhesion molecule-1 (VCAM-1) interactions have therapeutic potential in treating allergic airway disease because of the importance of these adhesion molecules in the trafficking of eosinophils, lymphocytes, and monocytes. We examined several small molecule inhibitors of alpha4beta1/VCAM-1 interactions with in vitro potencies (IC(50) values) ranging from 0.52 nM (CP-664511; 3-[3-(1-[2-[3-methoxy-4-(3-O-tolyl-ureido)phenyl]-acetylamino]-3-methyl-butyl)isoxazol-5-yl]-propionic acid) to 38.5 nM (CP-609643; 3-[3-methyl-1-[2-[4-(3-O-tolyl-ureido)-phenyl]-acetylamino]-butyl)-isoxazol-5-yl]-propionic acid). The same compounds were evaluated in vivo using a murine model of ovalbumin-induced pulmonary eosinophilia. In this model, systemic administration of antibodies against alpha4 reduced bronchoalveolar lavage (BAL) eosinophilia approximately 60%. Small molecule alpha4beta1 antagonists were administered by intratracheal instillation and demonstrated dose-dependent inhibition of BAL eosinophil numbers and achieved a maximum inhibition of approximately 60%. In general, the rank order of potency for these compounds in vitro was consistent with that observed in vivo, which confirms that their efficacy is likely via blockade of alpha4beta1/VCAM-1 interactions. The most potent compound, CP-664511, also inhibited BAL eosinophilia following s.c. administration (1-10 mg/kg, s.c.). These data support the utility of small molecule alpha4beta1 antagonists in the treatment of relevant diseases, such as asthma.
Asunto(s)
Antialérgicos/uso terapéutico , Integrinas/metabolismo , Isoxazoles/uso terapéutico , Compuestos de Fenilurea/uso terapéutico , Eosinofilia Pulmonar/tratamiento farmacológico , Receptores Mensajeros de Linfocitos/metabolismo , Molécula 1 de Adhesión Celular Vascular/metabolismo , Animales , Modelos Animales de Enfermedad , Humanos , Hipersensibilidad/etiología , Inflamación/etiología , Integrina alfa4beta1 , Integrinas/antagonistas & inhibidores , Células Jurkat , Ratones , Eosinofilia Pulmonar/complicaciones , Receptores Mensajeros de Linfocitos/antagonistas & inhibidoresRESUMEN
A series of isoxazolyl, oxazolyl, and thiazolylpropionic acid derivatives derived from LDV was found to be a potent antagonist of the alpha(4)beta(1) integrin. The synthesis and SAR leading up to 3-[3-(1-[-[3-methoxy-4-(3-o-tolyl-ureido)-phenyl]-acetylamino]-3-methyl-butyl)-isoxazol-5-yl]-propionic acid (22) are reported. In an allergic mouse model, compound 22 was efficacious delivered systemically (58% inhib @ 10 mg/kg, sc) as well as by intra-tracheal instillation (ED(50)=2 microg/kg).
Asunto(s)
Integrinas/antagonistas & inhibidores , Isoxazoles/farmacología , Propionatos/farmacología , Receptores Mensajeros de Linfocitos/antagonistas & inhibidores , Animales , Modelos Animales de Enfermedad , Humanos , Hipersensibilidad/tratamiento farmacológico , Integrina alfa4beta1 , Células Jurkat , Ratones , Relación Estructura-ActividadRESUMEN
p59fynT is a protein tyrosine kinase in the src family that has been associated with and believed to function in the signaling of many receptors, including the T-cell receptor. A role for the kinase in antigen-driven pulmonary inflammation was examined using mice whose p59fynT gene had been genetically ablated. FynKO mice that were sensitized to ovalbumin exhibited a marked increase in bronchoalveolar lavage eosinophils and cytokines, including interleukin (IL)-4 and IL-5, relative to wild-type mice in response to antigen aerosol exposure. Ovalbumin-stimulated IL-5 production was also increased in cultured splenocytes derived from fynKO mice relative to wild-type mice, whereas interferon-gamma levels were unchanged. Diminished concanavalin A--stimulated IL-4 levels from fynKO splenocytes were consistent with reduced serum immunoglobulin (Ig)E levels observed in sensitized/saline aerosol-challenged animals and may reflect defective natural killer 1.1(+) T cell development. Normalization of IgE levels in sensitized fynKO mice relative to wild-type mice occurred after repeat antigen challenge, which suggests a secondary source of IL-4. Overall, these data demonstrate fyn is a negative regulator of allergic airway inflammation in mice because its absence promotes a shift to a T helper-2 phenotype that may reflect the kinase's role in T-cell receptor signaling.
Asunto(s)
Hipersensibilidad/metabolismo , Neumonía/metabolismo , Proteínas Proto-Oncogénicas/genética , Familia-src Quinasas/genética , Animales , Antígenos/inmunología , Antígenos/farmacología , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Concanavalina A/farmacología , Modelos Animales de Enfermedad , Eosinófilos/inmunología , Hipersensibilidad/inmunología , Inmunoglobulina E/sangre , Interleucina-4/biosíntesis , Interleucina-5/biosíntesis , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ovalbúmina/inmunología , Ovalbúmina/farmacología , Neumonía/inmunología , Proteínas Proto-Oncogénicas/inmunología , Proteínas Proto-Oncogénicas c-fyn , Bazo/citología , Bazo/inmunología , Células Th2/inmunología , Células Th2/metabolismo , Familia-src Quinasas/inmunologíaRESUMEN
1. The ability of various C-C chemokines to elicit tissue eosinophil infiltration following intradermal injection or peripheral blood eosinophilia following intravenous injection were compared in the Brown-Norway rat. 2. Eotaxin (0.1 - 3 microg site-1) of human and murine origin produced equivalent, dose-dependent increases in eosinophil peroxidase activity in rat dermis 4 h post-injection. 3. Human eotaxin-2 was equipotent with human eotaxin in terms of dermal eosinophil recruitment. Other human CCR3 agonists, such as MCP-3, RANTES and MCP-4 failed to increase dermal eosinophil peroxidase activity at doses up to 1 microg site-1 whereas the latter did produce a small effect at 3 microg site-1. 4. Consistent with observations in vivo, human eotaxin displaced [125I]-eotaxin from rat spleen membranes more potently (IC50=2 nM) than did MCP-4 (IC50=500 nM). RANTES did not compete with the radiolabelled chemokine at concentrations up to 1 microM. 5. Human eotaxin (5 microg) administered intravenously increased circulating eosinophils approximately 3 fold whereas MCP-4 (5 microg i.v.) increased circulating monocytes approximately 3 fold without affecting eosinophil numbers. 6. Dexamethasone pretreatment inhibited eotaxin-induced dermal eosinophil influx only at a steroid dose (0.1 mg kg-1, s.c.) which significantly reduced circulating eosinophil numbers. The steroid also reduced eosinophilia in peripheral blood resulting from systemic eotaxin administration (5 microg, i.v.). 7. These data suggest differences in rat CCR3 relative to other species as surmised from a distinctive rank order of chemokine potency. In addition to its chemotactic effects eotaxin, but not MCP-4, promotes eosinophil recruitment into the circulation. One of the mechanisms by which glucocorticoids, such as dexamethasone, acutely inhibits eotaxin-induced dermal eosinophil influx is to diminish the circulating numbers of these cells available for tissue recruitment.
Asunto(s)
Quimiocinas CC , Eosinófilos/efectos de los fármacos , Receptores de Quimiocina/agonistas , Receptores del VIH/agonistas , Animales , Antiinflamatorios/farmacología , Agregación Celular/efectos de los fármacos , Quimiocina CCL11 , Quimiocinas/farmacología , Citocinas/farmacología , Dexametasona/farmacología , Eosinófilos/citología , Leucocitos/citología , Leucocitos/efectos de los fármacos , Ratas , Receptores CCR3RESUMEN
BACKGROUND: Histamine is a critical mediator of immediate hypersensitivity reactions. Sensory neuropeptides, such as substance P (SP), may also contribute to acute inflammatory responses. A compound which antagonizes both H1 and NK-1 receptors, such as MDL 108,207DA, may present a significant therapeutic advantage over pure antihistamines. METHODS: The binding affinity of MDL 108,207DA for H1 and NK-1 receptors was evaluated and its potency of antagonism evaluated in vitro. The in vivo antagonism of SP- or histamine-induced microvascular leakage in guinea pig airways was examined. A role for these mediators in antigen-induced microvascular leakage in ovalbumin-sensitized guinea pig airways was examined using MDL 108,207DA as well as the NK-1-selective antagonist FK888 and the H1-selective antagonist pyrilamine alone or in combination. RESULTS: The affinity of MDL 108,207DA for H1 and NK-1 receptors is similar to that of receptor-selective antagonists. The compound inhibits both receptors in vitro and in vivo with comparable potencies for each. The efficacy of FK888 in combination with pyrilamine and MDL 108,207DA on antigen-induced microvascular leakage in sensitized guinea pig airways supports a role for both SP and histamine in early allergic responses. CONCLUSION: The contribution of both SP and histamine to immediate hypersensitivity reactions supports the utility of NK-1 and H1 receptor antagonist therapy. MDL 108,207DA incorporates both activities into the same compound and, as a result, may be useful in the treatment of allergic diseases.
Asunto(s)
Bencimidazoles/farmacología , Antagonistas de los Receptores Histamínicos H1/farmacología , Histamina/fisiología , Antagonistas del Receptor de Neuroquinina-1 , Pirrolidinas/farmacología , Hipersensibilidad Respiratoria/fisiopatología , Sustancia P/fisiología , Animales , Bencimidazoles/metabolismo , Bronquios/irrigación sanguínea , Células CHO , Permeabilidad Capilar/efectos de los fármacos , Cricetinae , Dipéptidos/farmacología , Combinación de Medicamentos , Cobayas , Antagonistas de los Receptores Histamínicos H1/metabolismo , Indoles/farmacología , Fosfatos de Inositol/metabolismo , Contracción Muscular/efectos de los fármacos , Ovalbúmina , Pirilamina/farmacología , Pirrolidinas/metabolismo , Ensayo de Unión Radioligante , Receptores Histamínicos H1/metabolismo , Receptores de Neuroquinina-1/metabolismo , Hipersensibilidad Respiratoria/inducido químicamente , Hipersensibilidad Respiratoria/metabolismo , Pruebas Cutáneas , Tráquea/irrigación sanguínea , Células Tumorales Cultivadas/efectos de los fármacos , Células Tumorales Cultivadas/metabolismoRESUMEN
The tachykinins, substance P (SP) and neurokinin A (NK-A), are thought to be key players in the process of neurogenic inflammation, which is believed to contribute to the pathogenesis of various respiratory diseases. Due to the additive nature of the respiratory effects of these sensory neuropeptides, inhibiting the effects of tachykinins at both NK1 and NK2 receptors may represent a therapeutic advantage for the treatment of asthma, as opposed to receptor-selective antagonists, which have demonstrated only minimal efficacy to date. A number of companies are pursuing small molecule approaches yielding compounds with potent, balanced NK1 and NK2 receptor antagonist activities. In allergic rhinitis, NK1 receptor antagonism may complement the actions of antihistamines by addressing nasal congestion, which is largely unrelieved by these otherwise highly efficacious agents. Novel combined H1/NK1 receptor antagonists have been developed and may represent a therapeutic option for the treatment of this disease.
RESUMEN
1. Stimulation of sensory nerves causes release of tachykinins, including substance P (SP) and neurokinin A (NKA), which produce a variety of respiratory effects via NK-1 and NK-2 receptors, respectively. Hence, development of a compound which could potently and equivalently antagonize both receptors was pursued. 2. MDL 105,172A ((R)-1-[3-(3,4-dicholorophenyl)-1-(3,4,5-trimethoxybenzoyl)- 3-pyrrolidinyl]-4- phenyl-piperidine-4-morpholinecarboxamide) exhibited high affinity for NK-1 (4.34 nM) and NK-2 (2.05 nM) receptors. In vitro, the compound antagonized SP (pA2 = 8.36) or NKA (pA2 = 8.61)-induced inositol phosphate accumulation in tachykinin monoreceptor cell lines. 3. In anaesthetized guinea-pigs, MDL 105,172A inhibited SP-induced plasma protein extravasation (ED50 = 1 mg kg-1, i.v.) and [beta-Ala8]NKA 4-10-induced bronchoconstriction (ED50 = 0.5 mg kg-1, i.v.) indicating NK-1 and NK-2 antagonism, respectively. 4. Capsaicin was used to elicit respiratory effects in anaesthetized and conscious guinea-pigs; the latter were inhibited by MDL 105,172A following i.v. (ED50 = 1 mg kg-1) or oral (ED50 = 20 mg kg-1) administration. Hence, MDL 105,172A can inhibit pulmonary responses to tachykinins released endogenously in the airways. 5. At doses up to 200 mg kg-1, p.o., MDL 105,172A failed to inhibit repetitive hind paw tapping induced by i.c.v GR 73632, and NK-1 selective agonist, in gerbils, whereas CP-99,994 (0.87 mg kg-1, s.c.) completely ablated the effect. These data suggest that MDL 105,172A does not penetrate the central nervous system (CNS) and its tachykinin antagonism is restricted to the periphery. 6. MDL 105,172A is a non-peptide, potent, equivalent antagonist of NK-1 and NK-2 receptors. Its ability to inhibit both exogenously administered as well as endogenously released tachykinins support its use in examining the role of sensory neuropeptides in diseases associated with neurogenic inflammation including asthma.
Asunto(s)
Morfolinas/farmacología , Antagonistas del Receptor de Neuroquinina-1 , Piperidinas/farmacología , Receptores de Neuroquinina-2/antagonistas & inhibidores , Mecánica Respiratoria/efectos de los fármacos , Taquicininas/antagonistas & inhibidores , Animales , Conducta Animal/efectos de los fármacos , Permeabilidad Capilar/efectos de los fármacos , Gerbillinae , Cobayas , Técnicas In Vitro , Fosfatos de Inositol/fisiología , Masculino , Neuroquinina A/antagonistas & inhibidores , Neuroquinina A/farmacología , Fragmentos de Péptidos/farmacología , Pletismografía Total , Ensayo de Unión Radioligante , Sustancia P/análogos & derivados , Sustancia P/farmacología , Taquicininas/farmacologíaRESUMEN
A model of allergic pulmonary inflammation is described in which the intraperitoneal injection of antigen (Ag)-pulsed cells resulted in T cell priming. Mice received two injections of 10(6) elicited peritoneal macrophages, which had been incubated with Ag for 48 hr, on Days 0 and 6, followed by an aerosol Ag challenge on Day 19. Bronchoalveolar lavage fluid harvested on Day 21 contained increased eosinophil numbers and resembled the cell influx observed following immunization with Ag in alum. Incubation of Ag-presenting cells with interferon-gamma resulted in increased expression of the costimulator molecule B7-2 and of MHC Ags, but did not enhance priming capacity. Using this system, antibodies to CD4 and CD8 were tested for their ability to block sensitization by Ag-pulsed cells. Both anti-CD4 and anti-CD8 antibodies completely blocked the airway eosinophil response following aerosol Ag challenge. This model will be very useful for characterization of the interactions between Ag-presenting cells and T cells which ultimately result in the induction of pulmonary eosinophilia.
Asunto(s)
Células Presentadoras de Antígenos/inmunología , Asma/inmunología , Eosinofilia Pulmonar/etiología , Eosinofilia Pulmonar/inmunología , Linfocitos T/inmunología , Animales , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Modelos Animales de Enfermedad , Femenino , Citometría de Flujo , Ratones , Ratones Endogámicos C57BL , Ovalbúmina/inmunologíaRESUMEN
MDL 105,212 has been identified as a potent, nonpeptide NK-1 and NK-2 receptor antagonist that inhibits effects of substance P and neurokinin A in vitro and in vivo (Kudlacz et al., 1996). In the present study, the compound inhibited capsaicin-induced respiratory effects after p.o. administration (5-50 mg/kg) to conscious guinea pigs; nearly complete inhibition of dyspnea and cough was observed 1 hr after 50 mg/kg p.o., and efficacy persisted for approximately 11 hr. MDL 105,212 reduced pulmonary insufflation pressure and microvascular leakage in ovalbumin-sensitized animals in response to antigen-challenge relative to vehicle-treated animals. Attenuation of early-phase allergic responses may result from MDL 105,212 inhibition of antigen-induced histamine release from sensitized guinea pig lung observed in vitro. Airway hyperresponsiveness to methacholine occurred 24 hr after antigen-challenge in ovalbuminsensitized guinea pigs; this effect was inhibited by pretreatment with MDL 105,212 (50 mg/kg p.o.) 1 hr before ovalbumin exposure without affecting increased bronchoalveolar lavage eosinophil numbers. These data suggest that sensory neuropeptides play a role in some aspects of allergic airway responses and that tachykinin receptor antagonists may be useful in treatment of atopic respiratory diseases.
Asunto(s)
Hipersensibilidad/tratamiento farmacológico , Antagonistas del Receptor de Neuroquinina-1 , Piperidinas/farmacología , Pirrolidinas/farmacología , Receptores de Neuroquinina-2/antagonistas & inhibidores , Animales , Broncoconstricción/efectos de los fármacos , Capsaicina/farmacología , Relación Dosis-Respuesta a Droga , Cobayas , Liberación de Histamina/efectos de los fármacos , Masculino , Ovalbúmina/inmunología , Respiración/efectos de los fármacosRESUMEN
We have identified and characterized a novel, potent, nonselective tachykinin receptor antagonist, MDL 105,212A [(R)-1-[2-[3-(3,4- dichlorophenyl)-1-(3,4,5-trimethoxybenzoyl)-pyrrolidin-3-yl] -ethyl]- 4-phenylpiperidine-4-carboxamide, hydrochloride]. The compound binds with low nanomolar affinity and species specificity to human NK-1 and NK-2 receptors as well as to guinea pig NK-3 receptors. In vitro functional assays are consistent with potent competitive antagonism of substance P-(SP) or neurokinin A-(NKA) induced [3H]-inositol phosphate accumulation in NK-1 or NK-2 monoreceptor cell lines with pA2 values of 8.19 and 8.67, respectively. Its ability to inhibit SP, NKA and capsaicin-mediated respiratory effects was examined in guinea pigs in vivo. MDL 105,212A attenuated SP-induced airway plasma protein extravasation (ED50 = 0.20 mg/kg, i.v.), NKA-induced respiratory collapse (ED50 = 5 mg/kg, i.v) and inhibited capsaicin-induced increases in pulmonary insufflation pressure (ED50 = 0.5 mg/kg, i.v.). Conscious guinea pigs responded to capsaicin aerosol exposure with dyspnea, coughs and gasps (significant respiratory events) and plasma protein extravasation. MDL 105,212A inhibited these responses in a dose-dependent manner after i.v. (ED50 = 5 mg/kg) or oral (ED50 = 50 mg/kg) administration. These data suggest that MDL 105,212A is a potent NK-1 and NK-2 receptor antagonist based on in vitro activity and its ability to inhibit SP and NKA mediated respiratory effects in vivo after exogenous administration or endogenous release and hence may be a useful therapeutic agent in neuroinflammatory disorders such as asthma in which a role for both tachykinins in the pathogenesis of the disease has been postulated.
Asunto(s)
Antagonistas del Receptor de Neuroquinina-1 , Piperidinas/farmacología , Pirrolidinas/farmacología , Receptores de Neuroquinina-2/antagonistas & inhibidores , Secuencia de Aminoácidos , Animales , Asma/tratamiento farmacológico , Broncoconstricción/efectos de los fármacos , Permeabilidad Capilar/efectos de los fármacos , Cobayas , Humanos , Fosfatos de Inositol/metabolismo , Masculino , Cloruro de Metacolina/farmacología , Ratones , Datos de Secuencia Molecular , Neuroquinina A/antagonistas & inhibidores , Ratas , Respiración/efectos de los fármacos , Especificidad de la Especie , Sustancia P/antagonistas & inhibidoresRESUMEN
Respiratory viral infections not only exacerbate asthma symptoms but may also be important in the pathogenesis of the disease. We therefore explored the effects of respiratory viral infection on the respiratory response of sensitized guinea pigs to antigen challenge. Lung tissue obtained from uninfected guinea pigs sensitized to ovalbumin aerosol released histamine upon incubation with the antigen in vitro. After antigen challenge in vivo, sensitized animals had significantly greater numbers of eosinophils in their bronchoalveolar lavage fluid than did nonsensitized animals and exhibited airway hyperresponsiveness to methacholine aerosol. When ovalbumin sensitization was initiated 7 days after inoculation with parainfluenza virus type-3 (PI-3), antigen challenge elicited little histamine release from infected lung tissue in vitro. Likewise, subsequent to antigen challenge in vivo, animals failed to exhibit airway hyperresponsiveness or an increased eosinophil population in bronchoalveolar lavage fluid. Similar effects were observed when sensitization was begun 19 days after PI-3 virus inoculation. The mechanism(s) responsible for the attenuated responses to antigen in PI-3 infected animals are unknown but may involve virus-induced effects on immune cells.
Asunto(s)
Hiperreactividad Bronquial/inmunología , Virus de la Parainfluenza 3 Humana/inmunología , Infecciones por Paramyxoviridae/inmunología , Aerosoles , Animales , Líquido del Lavado Bronquioalveolar/citología , Recuento de Células/efectos de los fármacos , Modelos Animales de Enfermedad , Disnea/diagnóstico , Disnea/inmunología , Eosinófilos/citología , Eosinófilos/efectos de los fármacos , Cobayas , Liberación de Histamina/efectos de los fármacos , Liberación de Histamina/inmunología , Técnicas In Vitro , Masculino , Cloruro de Metacolina/administración & dosificación , Cloruro de Metacolina/toxicidad , Ovalbúmina/inmunología , Ovalbúmina/toxicidad , Virus de la Parainfluenza 3 Humana/efectos de los fármacosRESUMEN
We have investigated the effects of parainfluenza virus type 3 (PI-3) on sensory neuropeptide levels, tachykinin receptors and their functions in guinea pig airways during the course of respiratory viral infection. PI-3 infected guinea pigs were hyperresponsive to methacholine and substance P aerosols as determined by earlier onset of dyspnea in these animals as compared with control on post-inoculation day (PID) 7 but not 19. In addition, plasma protein extravasation produced in response to the tachykinin was increased in infected airways during the first week post inoculation. Infected guinea pig trachea did not respond any differently to methacholine when smooth muscle contraction and [3H]inositol phosphate accumulation were measured although the magnitude of substance P effects using in vitro tests was significantly greater than control on post-inoculation day 7 but not 19. Trachea from PI-3 infected animals were characterized by reductions in substance P-like immunoreactivity, tachykinin NK1 receptor number and agonist affinity during the first post-inoculation week. Substance P levels or tachykinin NK1 receptor numbers or affinity were not altered in trachea of guinea pigs 4 days after treatment with lipopolysaccharide. These data suggest substance P release occurs during critical periods of respiratory viral infection which are temporally correlated with airway hyperresponsiveness. Despite apparent down-regulation of tachykinin NK1 receptors, substance P-mediated functions remained enhanced suggesting some alterations in post-receptor mechanisms.
Asunto(s)
Virus de la Parainfluenza 3 Humana , Infecciones por Paramyxoviridae/fisiopatología , Receptores de Neuroquinina-1/metabolismo , Mecánica Respiratoria/fisiología , Sustancia P/metabolismo , Tráquea/fisiopatología , Animales , Proteínas Sanguíneas/metabolismo , Permeabilidad Capilar/fisiología , Escherichia coli , Cobayas , Técnicas In Vitro , Fosfatos de Inositol/metabolismo , Lipopolisacáridos/farmacología , Masculino , Cloruro de Metacolina/farmacología , Músculo Liso/fisiología , Neuronas Aferentes/metabolismo , Neuronas Aferentes/fisiología , Neuropéptidos/metabolismo , Infecciones por Paramyxoviridae/metabolismo , Ensayo de Unión Radioligante , Sustancia P/farmacología , Sinapsis/fisiología , Tráquea/inervación , Tráquea/metabolismoRESUMEN
The effects of sensory neuropeptides substance P (SP) and neurokinin A (NKA) on immune cell recruitment and macrophage activation were determined. Guinea pigs exposed to capsaicin aerosol exhibited eosinophil and neutrophil influx into their bronchoalveolar lavage (BAL) fluid 24 h after treatment; SP aerosol elicited eosinophil influx, whereas NKA aerosol exposure caused neutrophil recruitment. Inhalation of capsaicin, NKA or SP aerosols also enhanced superoxide production induced by zymosan in cultured alveolar macrophages. Incubation of alveolar macrophages with SP or NKA in culture for the same time (24 h) did not potentiate the response to zymosan. Hence, tachykinin-mediated airway effects may not be the result of direct actions on target cells but rather involve alternate mechanisms and mediators which do not necessarily reflect in vitro data.
Asunto(s)
Bronquios/efectos de los fármacos , Sistema Inmunológico/efectos de los fármacos , Alveolos Pulmonares/efectos de los fármacos , Taquicininas/farmacología , Aerosoles , Animales , Aniones/metabolismo , Líquido del Lavado Bronquioalveolar/citología , Capsaicina/farmacología , Cobayas , Sistema Inmunológico/citología , Sistema Inmunológico/fisiología , Macrófagos Alveolares/metabolismo , Masculino , Superóxidos/metabolismoRESUMEN
The study of virus-induced airway hyperresponsiveness may provide insight into mechanisms that contribute to respiratory diseases such as asthma. We examined changes induced by parainfluenza virus type 3 (PI-3) in lung lesions, tissue weights, and airway responsiveness to aerosols of histamine, methacholine, or citric acid in conscious guinea pigs, using modified whole body plethysmography. During the first week after inoculation, infected lung tissue had peribronchiolitis and airway hyperresponsiveness to various agents when dyspnea and significant respiratory events were measured; these effects persisted throughout postinoculation weeks 2 and 3. Airway hyperresponsiveness was defined by reductions in the onset of dyspnea or significant respiratory events. Throughout the course of the study, PI-3 infected animals had resting respiratory patterns that reflected labored breathing and may have been related to the edema indicated by increased lung weights. Furthermore, increased numbers of inflammatory cells were observed in lung tissue as well as bronchoalveolar lavage fluid of infected animals at these times. Unlike PI-3 infection, exposure to gram-negative endotoxin resulted primarily in airway hyporesponsiveness to histamine aerosol. Hence, we have shown PI-3 infection in guinea pigs causes time-dependent alterations in airway responsiveness to diverse bronchoactive agents as well as in normal breathing patterns, which may persist up to several weeks after inoculation in animals that may otherwise appear normal.
Asunto(s)
Pulmón/patología , Virus de la Parainfluenza 3 Humana , Infecciones por Paramyxoviridae/patología , Hipersensibilidad Respiratoria/fisiopatología , Animales , Líquido del Lavado Bronquioalveolar/citología , Modelos Animales de Enfermedad , Endotoxinas/efectos adversos , Cobayas , Lipopolisacáridos/efectos adversos , Pulmón/fisiopatología , Masculino , Infecciones por Paramyxoviridae/complicaciones , Infecciones por Paramyxoviridae/fisiopatología , Hipersensibilidad Respiratoria/inducido químicamente , Hipersensibilidad Respiratoria/complicaciones , Factores de Tiempo , Aumento de PesoRESUMEN
Tachykinins, in particular neurokinin A and substance P, produce a number of airway effects which may contribute to respiratory diseases such as asthma. We examined the ability of aerosolized substance P, neurokinin A or capsaicin to produce respiratory alterations in conscious guinea pigs using modified whole body plethysmography. Substance P-mediated dyspnea and significant respiratory events were inhibited by the NK1 receptor antagonist, CP-96,345. Neurokinin A-mediated respiratory effects were ablated by the NK2 receptor antagonists: MEN 10207, MDL 29,913 and SR 48,968, the latter being the most potent. The peptide-based antagonist, MEN 10207, produced respiratory effects itself suggesting partial agonist activity. The cyclic hexapeptide, MDL 29,913, relaxed airway smooth muscle via mechanisms other than tachykinin antagonism. NK2 but not NK1 receptor antagonists were able to delay the onset of capsaicin-induced dyspnea, although alone they did not usually (in approximately 10% of the animals) eliminate the response. However, when NK2 receptor antagonists were combined with CP-96,345, the incidence of dyspnea induced by capsaicin decreased significantly (40%) suggesting that both tachykinins contribute to dyspnea in this system.
Asunto(s)
Antagonistas del Receptor de Neuroquinina-1 , Receptores de Neuroquinina-2/antagonistas & inhibidores , Respiración/efectos de los fármacos , Taquicininas/farmacología , Aerosoles , Animales , Compuestos de Bifenilo/farmacología , Capsaicina/farmacología , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Cobayas , Hipnóticos y Sedantes/farmacología , Técnicas In Vitro , Masculino , Neuroquinina A/farmacología , Neuropéptidos/farmacología , Sustancia P/farmacología , Taquicininas/antagonistas & inhibidoresRESUMEN
Early exposure of developing postsynaptic receptors to the appropriate neurotransmitter is thought to be important for establishing the pattern of receptor development and function. We have examined the role of fetal catecholamines in central nervous system beta-adrenergic receptor maturation by administering the beta-adrenergic antagonist, propranolol, throughout gestation via continuous maternal infusion in rats. After birth, we examined beta-receptor binding of [125I]pindolol in three brain regions. In the cerebral cortex, receptor binding was relatively normal in the propranolol group during the immediate postpartum period, but eventually became elevated in young adulthood. Binding characteristics were not significantly affected in midbrain + brainstem or cerebellum. These data support a role for fetal catecholamines in the programming of subsequent receptor development, but in a fashion selective for specific brain regions.
Asunto(s)
Encéfalo/metabolismo , Propranolol/farmacología , Receptores Adrenérgicos beta/metabolismo , Animales , Peso Corporal/efectos de los fármacos , Encéfalo/efectos de los fármacos , Encéfalo/crecimiento & desarrollo , Tronco Encefálico/efectos de los fármacos , Tronco Encefálico/metabolismo , Cerebelo/efectos de los fármacos , Cerebelo/metabolismo , Femenino , Infusiones Intravenosas , Intercambio Materno-Fetal , Membranas/metabolismo , Tamaño de los Órganos/efectos de los fármacos , Pindolol/metabolismo , Embarazo , Efectos Tardíos de la Exposición Prenatal , Propranolol/administración & dosificación , Ratas , Ratas Endogámicas , Regulación hacia Arriba/efectos de los fármacosRESUMEN
The role of prenatal beta-receptor stimulation in development of adrenergic reactivity was examined by administering the beta-agonist, terbutaline, to pregnant rats on gestational days 17, 18 and 19. On gestational day 20, liver membrane beta-receptor binding capabilities showed the depression characteristic of down-regulation, but heart and kidney receptor binding were essentially normal. Basal adenylate cyclase activity in the fetal liver membrane preparation was unchanged by terbutaline exposure and enzymatic reactivity to beta-adrenergic stimulation showed only a slight lowering; forskolin stimulation, however, was markedly increased in the terbutaline group. By postnatal day 2, receptor binding had returned to normal in the liver and remained at control levels in the other two tissues. Responsivity of adenylate cyclase to beta-receptor stimulation was markedly elevated in heart and kidney membranes; the effect represented an alteration at the level of the cyclase itself, rather than the receptor, since both basal activity and forskolin stimulation of the enzyme showed equivalent enhancement. These data thus suggest that early beta-adrenergic stimulation promotes cellular reactivity by fostering the development of membrane transduction mechanisms, rather than through effects on the receptor ligand binding site per se.
Asunto(s)
Efectos Tardíos de la Exposición Prenatal , Terbutalina/farmacología , Adenilil Ciclasas/metabolismo , Agonistas Adrenérgicos beta/farmacología , Envejecimiento/metabolismo , Animales , Peso Corporal/efectos de los fármacos , Colforsina/farmacología , Regulación hacia Abajo , Femenino , Tamaño de la Camada/efectos de los fármacos , Hígado/enzimología , Tamaño de los Órganos/efectos de los fármacos , Embarazo , Ratas , Ratas Endogámicas , Receptores Adrenérgicos beta/metabolismo , Receptores Adrenérgicos beta/fisiología , Distribución TisularRESUMEN
Adrenergic stimulation and glucocorticoids have been hypothesized to control the development of beta-adrenergic receptors and responsiveness. In the current study, rats were exposed to a beta-agonist (terbutaline) or a glucocorticoid (dexamethasone) during late gestation, and the development of adenylate cyclase activity and beta-receptor binding was evaluated in membranes prepared from the heart and liver. In control heart, basal, isoproterenol-stimulated and forskolin-stimulated adenylate cyclase showed distinct developmental spikes of activity that were unrelated to changes in receptor binding sites, but that instead corresponded temporally to periods of sympathetic neuronal activation. Prenatal exposure to terbutaline initially enhanced all three enzymatic measures in the immediate postpartum period but delayed the appearance and exaggerated the magnitude of the subsequent peaks; again, these changes occurred without specific relation to effects on receptor binding. Dexamethasone produced a similar shift in the peaks of cardiac enzyme activity. In the liver, where beta-adrenergic receptors and responsiveness decline after birth, terbutaline and dexamethasone had much smaller effects on adenylate cyclase. These results suggest that beta-adrenergic stimulation serves as a trophic factor controlling the ontogenetic rise of adenylate cyclase activity; regulation involves the level of the enzyme itself rather than changes in receptor binding capabilities or receptor-specific linkages. Consequently, prenatal administration of beta-agonists or glucocorticoids can cause long-term alterations in enzyme activity and responsiveness.
Asunto(s)
Adenilil Ciclasas/metabolismo , Dexametasona/farmacología , Corazón/crecimiento & desarrollo , Hígado/crecimiento & desarrollo , Miocardio/enzimología , Efectos Tardíos de la Exposición Prenatal , Terbutalina/farmacología , Animales , Animales Recién Nacidos/metabolismo , Membrana Celular/enzimología , Colforsina/farmacología , Femenino , Corazón/efectos de los fármacos , Corazón/embriología , Isoproterenol/farmacología , Hígado/embriología , Hígado/enzimología , Embarazo , Ratas , Ratas Endogámicas , Receptores Adrenérgicos beta/metabolismoRESUMEN
beta-Adrenergic receptor stimulation is thought to participate in the perinatal switchover of the lung to air-breathing. In the current study, we have determined whether prenatal exposure of rats to terbutaline (10 mg/kg on gestational days 17,18 and 19) exerts promotional effects on lung function solely through its immediate actions at beta-receptors or whether terbutaline influences subsequent reactivity to adrenergic stimuli; we have contrasted the effects with those seen with a glucocorticoid (dexamethasone, 0.8 mg/kg on gestational days 17, 18 and 19). On postnatal day 3, basal lung compliance and lung rupture volumes were reduced after treatment with either terbutaline or dexamethasone, effects related primarily to drug-induced growth retardation; these effects were not present if data were corrected for relative tissue size. Despite a slightly reduced basal compliance, prenatal terbutaline exposure markedly enhanced postnatal reactivity to acute challenge with isoproterenol. In contrast, dexamethasone primarily enhanced basal compliance (corrected for weight deficits), with only minor effects on the isoproterenol response. These results suggest that, rather than producing classical receptor desensitization, gestational exposure to a beta-agonist sensitizes lung compliance to postnatal beta-receptor stimulation. Promotional effects of terbutaline on neonatal lung function are thus distinct from functional changes achieved with glucocorticoids, such as dexamethasone.