RESUMEN
The expression of artemin (ARTN), a glial cell line-derived neurotrophic factor (GDNF) family ligand, increases in pre-clinical models of nociception and recent evidence suggests this growth factor may play a causative role in inflammatory pain mechanisms. The aim of this study was to demonstrate functional inhibition of ARTN with monoclonal antibodies and to determine whether ARTN neutralisation could reverse inflammatory pain in mice. We show that monoclonal antibodies with high affinity to ARTN, completely inhibit ARTN-induced Ret and ERK activation in a human neuroblastoma cell line, and block capsaicin-induced CGRP secretion from primary rat DRG cultures. In addition, administration of anti-ARTN antibodies to mice provides a transient, partial reversal (41%) of FCA-induced mechanical hypersensitivity. Anti-ARTN antibodies had no effect on hypersensitivity in response to partial nerve ligation in mice. These data suggest that ARTN-GFRα3 interactions partially mediate early stage nociceptive signalling following an inflammatory insult.
Asunto(s)
Ganglios Espinales/metabolismo , Receptores del Factor Neurotrófico Derivado de la Línea Celular Glial/metabolismo , Hiperalgesia/fisiopatología , Proteínas del Tejido Nervioso/metabolismo , Transducción de Señal , Animales , Calor , Masculino , Unión Proteica , Ratas , Ratas Sprague-DawleyRESUMEN
Animal models are an integral part of pain research. However, current models tend to rely on evoked responses and there is a belief that non-evoked responses may be a more relevant behavioural readout as the animal responds in a more natural manner. Here, dynamic weight bearing (DWB), a novel method for assessing mechanical hypersensitivity, was evaluated using the Freund's Complete Adjuvant (FCA) model of inflammatory pain in mice. DWB enables the measurement of weight placed through all four paws of a freely moving animal. The data obtained from DWB was compared with data acquired using the standard static weight bearing (incapacitance) test. In both tests reversal of FCA induced mechanical hypersensitivity was investigated using the selective COX2 inhibitor celecoxib. Mice treated with FCA placed less weight through the ipsilateral hindpaw compared to vehicle controls. This reduction was reversed by celecoxib (30mg/kg p.o.) in the dynamic and static weight bearing tests. The data presented here suggests that dynamic weight bearing may provide a novel end point for the development of new analgesics.
Asunto(s)
Adyuvantes Inmunológicos , Adyuvante de Freund , Hiperalgesia/fisiopatología , Soporte de Peso , Animales , Celecoxib , Dolor Crónico/etiología , Dolor Crónico/fisiopatología , Inhibidores de la Ciclooxigenasa 2/farmacología , Determinación de Punto Final , Femenino , Hiperalgesia/etiología , Inflamación/etiología , Inflamación/fisiopatología , Ratones , Ratones Endogámicos C57BL , Dimensión del Dolor , Pirazoles/farmacología , Sulfonamidas/farmacologíaRESUMEN
GSK207040 (5-[(3-cyclobutyl-2,3,4,5-tetrahydro-1H-3-benzazepin-7-yl)oxy]-N-methyl-2-pyrazinecarboxamide) and GSK334429 (1-(1-methylethyl)-4-({1-[6-(trifluoromethyl)-3-pyridinyl]-4-piperidinyl}carbonyl)hexahydro-1H-1,4-diazepine) are novel and selective non-imidazole histamine H(3) receptor antagonists from distinct chemical series with high affinity for human (pK(i)=9.67+/-0.06 and 9.49+/-0.09, respectively) and rat (pK(i)=9.08+/-0.16 and 9.12+/-0.14, respectively) H(3) receptors expressed in cerebral cortex. At the human recombinant H(3) receptor, GSK207040 and GSK334429 were potent functional antagonists (pA(2)=9.26+/-0.04 and 8.84+/-0.04, respectively versus H(3) agonist-induced changes in cAMP) and exhibited inverse agonist properties (pIC(50)=9.20+/-0.36 and 8.59+/-0.04 versus basal GTPgammaS binding). Following oral administration, GSK207040 and GSK334429 potently inhibited cortical ex vivo [(3)H]-R-alpha-methylhistamine binding (ED(50)=0.03 and 0.35 mg/kg, respectively). Functional antagonism of central H(3) receptors was demonstrated by blockade of R-alpha-methylhistamine-induced dipsogenia in rats (ID(50)=0.02 and 0.11 mg/kg p.o. for GSK207040 and GSK334429, respectively). In more pathophysiologically relevant pharmacodynamic models, GSK207040 (0.1, 0.3, 1 and 3mg/kg p.o.) and GSK334429 (0.3, 1 and 3mg/kg p.o.) significantly reversed amnesia induced by the cholinergic antagonist scopolamine in a passive avoidance paradigm. In addition, GSK207040 (0.1, 0.3 and 1mg/kg p.o.) and GSK334429 (3 and 10mg/kg p.o.) significantly reversed capsaicin-induced reductions in paw withdrawal threshold, suggesting for the first time that blockade of H(3) receptors may be able to reduce tactile allodynia. Novel H(3) receptor antagonists such as GSK207040 and GSK334429 may therefore have therapeutic potential not only in dementia but also in neuropathic pain.
Asunto(s)
Azepinas/uso terapéutico , Benzazepinas/uso terapéutico , Capsaicina , Antagonistas de los Receptores Histamínicos/uso terapéutico , Trastornos de la Memoria/tratamiento farmacológico , Neuralgia/tratamiento farmacológico , Pirazinas/uso terapéutico , Piridinas/uso terapéutico , Receptores Histamínicos H3/metabolismo , Escopolamina , Analgésicos/farmacocinética , Analgésicos/farmacología , Analgésicos/uso terapéutico , Animales , Reacción de Prevención/efectos de los fármacos , Azepinas/administración & dosificación , Azepinas/farmacocinética , Benzazepinas/farmacocinética , Benzazepinas/farmacología , Sistema Nervioso Central/efectos de los fármacos , Ingestión de Líquidos/efectos de los fármacos , Agonistas de los Receptores Histamínicos/farmacocinética , Agonistas de los Receptores Histamínicos/farmacología , Antagonistas de los Receptores Histamínicos/farmacocinética , Antagonistas de los Receptores Histamínicos/farmacología , Humanos , Masculino , Trastornos de la Memoria/inducido químicamente , Neuralgia/inducido químicamente , Pirazinas/farmacocinética , Pirazinas/farmacología , Piridinas/administración & dosificación , Piridinas/farmacocinética , Ratas , Ratas Sprague-DawleyRESUMEN
1. Two P2X(3)/P2X(2/3) receptor antagonists with different potencies were profiled electrophysiologically in a rat model of nerve injury. 2. A-317491 has poor CNS penetrance (blood:brain, 1:<0.05), and was therefore administered intravenously in chronic constriction injury (CCI)- and sham-operated rats to study the involvement of P2X(3) subunit-containing receptors in the periphery in neuropathic pain. A-317491 and Compound A were administered topically to the spinal cord to investigate the central contribution. 3. There were no significant inhibitory effects of A-317491 intravenous (i.v.) seen in sham-operated animals compared to vehicle controls. In CCI-operated animals, there were significant inhibitory effects of 3 mg kg(-1) A-317491 i.v. on C fibre-evoked responses, and with 10 mg kg(-1) A-317491 i.v. on A delta and C fibre-evoked responses. No significant effects of A-317491 were observed after topical application to the spinal cord. In contrast, when Compound A was administered spinally in CCI animals, there was a decrease in A delta and C fibre-evoked responses, and wind up. 4. These changes indicate that A-317491 has a selective effect on neuronal responses in CCI animals compared to sham, demonstrating an increased involvement of P2X(3)/P2X(2/3) receptors in sensory signalling following nerve injury. In addition, the more potent antagonist Compound A was effective spinally, unmasking a potential central role of P2X(3)/P2X(2/3) receptors at this site post nerve injury. These data support a role for P2X(3)/P2X(2/3) antagonists in the modulation of neuropathic pain.
Asunto(s)
Neuralgia/etiología , Receptores Purinérgicos P2/fisiología , Animales , Constricción Patológica , Potenciales Evocados/efectos de los fármacos , Masculino , Fibras Nerviosas Amielínicas/efectos de los fármacos , Fibras Nerviosas Amielínicas/fisiología , Neuralgia/tratamiento farmacológico , Neuralgia/fisiopatología , Fenoles/farmacocinética , Fenoles/farmacología , Compuestos Policíclicos/farmacocinética , Compuestos Policíclicos/farmacología , Ratas , Receptores Purinérgicos P2X2 , Receptores Purinérgicos P2X3 , Médula Espinal/efectos de los fármacos , Médula Espinal/fisiologíaRESUMEN
Small molecule antagonists of the vanilloid receptor TRPV1 (also known as VR1) are disclosed. Pyrrolidinyl ureas such as 8 and 15 (SB-705498) emerged as lead compounds following optimisation of the previously described urea SB-452533. Pharmacological studies using electrophysiological and FLIPR-Ca2+-based assays showed that compounds such as 8 and 15 were potent antagonists versus the multiple chemical and physical modes of TRPV1 activation (namely capsaicin, acid and noxious heat). Furthermore, 15 possessed suitable developability properties to enable progression of this compound into in vivo studies and subsequently clinical development.
Asunto(s)
Pirrolidinas/síntesis química , Pirrolidinas/farmacología , Canales Catiónicos TRPV/antagonistas & inhibidores , Urea/análogos & derivados , Administración Oral , Animales , Capsaicina/farmacología , Línea Celular , Diseño de Fármacos , Cobayas , Humanos , Estructura Molecular , Pirrolidinas/administración & dosificación , Pirrolidinas/química , Ratas , Relación Estructura-Actividad , Canales Catiónicos TRPV/metabolismo , Urea/administración & dosificación , Urea/síntesis química , Urea/química , Urea/farmacologíaRESUMEN
The pathogenic form of the cyclooxygenase (COX) enzyme, COX-2, is also constitutively present in the spinal cord and has been implicated in chronic pain states in rat and man. A number of COX-2 inhibitors, including celecoxib and rofecoxib, are already used in man for the treatment of inflammatory pain. Preclinically, the dual-acting COX-2 inhibitor, GW406381X [2-(4-ethoxyphenyl)-3-[4-(methylsulfonyl)phenyl]-pyrazolo[1,5-b]pyridazine, where X denotes the free base], is as effective as rofecoxib and celecoxib in the rat established Freund's Complete Adjuvant model with an ED(50) of 1.5 mg/kg p.o. compared with 1.0 mg/kg p.o. for rofecoxib and 6.6 mg/kg p.o. for celecoxib. However, in contrast to celecoxib (5 mg/kg p.o. b.i.d.) and rofecoxib (5 mg/kg p.o. b.i.d.), which were without significant effect, GW406381X (5 mg/kg p.o. b.i.d.) fully reversed mechanical allodynia in the chronic constriction injury model and reversed thermal hyperalgesia in the mouse partial ligation model, both models of neuropathic pain. GW406381X, was also effective in a rat model of capsaicin-induced central sensitization, when given intrathecally (ED(50) = 0.07 mug) and after chronic but not acute oral dosing. Celecoxib and rofecoxib had no effect in this model. Several hypotheses have been proposed to try to explain these differences in efficacy, including central nervous system penetration, enzyme kinetics, and potency. The novel finding of effectiveness of GW406381X in these models of neuropathic pain/central sensitization, in addition to activity in inflammatory pain models and together with its central efficacy, suggests dual activity of GW406381X compared with celecoxib and rofecoxib, which may translate into greater efficacy in a broader spectrum of pain states in the clinic.