RESUMO
Although opiates represent the most effective analgesics, their use in chronic treatments is associated with numerous side effects including the development of pain hypersensitivity and analgesic tolerance. We recently identified a novel orally active neuropeptide FF (NPFF) receptor antagonist, RF313, which efficiently prevents the development of fentanyl-induced hyperalgesia in rats. In this study, we investigated the properties of this compound into more details. We show that RF313 exhibited a pronounced selectivity for NPFF receptors, antagonist activity at NPFF1 receptor (NPFF1R) subtype both in vitro and in vivo and no major side effects when administered in mice up to 30 mg/kg. When co-administered with opiates in rats and mice, it improved their analgesic efficacy and prevented the development of long lasting opioid-induced hyperalgesia. Moreover, and in marked contrast with the dipeptidic NPFF receptor antagonist RF9, RF313 displayed negligible affinity and no agonist activity (up to 100 µM) toward the kisspeptin receptor. Finally, in male hamster, RF313 had no effect when administered alone but fully blocked the increase in LH induced by RFRP-3, while RF9 per se induced a significant increase in LH levels which is consistent with its ability to activate kisspeptin receptors. Altogether, our data indicate that RF313 represents an interesting compound for the development of therapeutic tools aiming at improving analgesic action of opiates and reducing adverse side effects associated with their chronic administration. Moreover, its lack of agonist activity at the kisspeptin receptor indicates that RF313 might be considered a better pharmacological tool, when compared to RF9, to examine the regulatory roles of RF-amide-related peptides and NPFF1R in reproduction.
Assuntos
Analgésicos Opioides/uso terapêutico , Hiperalgesia/tratamento farmacológico , Antagonistas de Entorpecentes/uso terapêutico , Oligopeptídeos/uso terapêutico , Receptores de Neuropeptídeos/antagonistas & inibidores , Administração Oral , Animais , Células CHO , Cricetinae , Cricetulus , Modelos Animais de Doenças , Fentanila/farmacologia , Humanos , Masculino , Mesocricetus , Camundongos , Camundongos Endogâmicos C57BL , Oligopeptídeos/química , Peptídeos/uso terapêutico , Piperidinas/química , Piperidinas/uso terapêutico , Ligação Proteica/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Receptores de Neuropeptídeos/metabolismo , Valina/análogos & derivados , Valina/química , Valina/uso terapêuticoRESUMO
Mutations of the AMP-activated kinase gamma 2 subunit (AMPKγ2), N488I (AMPKγ2NI) and R531G (AMPKγ2RG), are associated with Wolff-Parkinson-White (WPW) syndrome, a cardiac disorder characterized by ventricular pre-excitation in humans. Cardiac-specific transgenic overexpression of human AMPKγ2NI or AMPKγ2RG leads to constitutive AMPK activation and the WPW phenotype in mice. However, overexpression of these mutant proteins also caused profound, non-physiological increase in cardiac glycogen, which might abnormally alter the true phenotype. To investigate whether physiological levels of AMPKγ2NI or AMPKγ2RG mutation cause WPW syndrome and metabolic changes in other organs, we generated two knock-in mouse lines on the C57BL/6N background harboring mutations of human AMPKγ2NI and AMPKγ2RG, respectively. Similar to the reported phenotypes of mice overexpressing AMPKγ2NI or AMPKγ2RG in the heart, both lines developed WPW syndrome and cardiac hypertrophy; however, these effects were independent of cardiac glycogen accumulation. Compared with AMPKγ2WT mice, AMPKγ2NI and AMPKγ2RG mice exhibited reduced body weight, fat mass, and liver steatosis when fed with a high fat diet (HFD). Surprisingly, AMPKγ2RG but not AMPKγ2NI mice fed with an HFD exhibited severe kidney injury characterized by glycogen accumulation, inflammation, apoptosis, cyst formation, and impaired renal function. These results demonstrate that expression of AMPKγ2NI and AMPKγ2RG mutations at physiological levels can induce beneficial metabolic effects but that this is accompanied by WPW syndrome. Our data also reveal an unexpected effect of AMPKγ2RG in the kidney, linking lifelong constitutive activation of AMPK to a potential risk for kidney dysfunction in the context of an HFD.
Assuntos
Proteínas Quinases Ativadas por AMP/genética , Mutação , Insuficiência Renal/genética , Síndrome de Wolff-Parkinson-White/genética , Animais , Apoptose , Modelos Animais de Doenças , Técnicas de Introdução de Genes , Inflamação/genética , Inflamação/patologia , Rim/metabolismo , Rim/patologia , Masculino , Camundongos Endogâmicos C57BL , Insuficiência Renal/patologia , Síndrome de Wolff-Parkinson-White/patologiaRESUMO
Through the development of a new class of unnatural ornithine derivatives as bioisosteres of arginine, we have designed an orally active peptidomimetic antagonist of neuropeptide FF receptors (NPFFR). Systemic low-dose administration of this compound to rats blocked opioid-induced hyperalgesia, without any apparent side-effects. Interestingly, we also observed that this compound potentiated opioid-induced analgesia. This unnatural ornithine derivative provides a novel therapeutic approach for both improving analgesia and reducing hyperalgesia induced by opioids in patients being treated for chronic pain.
Assuntos
Analgésicos Opioides/toxicidade , Fentanila/toxicidade , Hiperalgesia/induzido quimicamente , Hiperalgesia/prevenção & controle , Peptidomiméticos/uso terapêutico , Animais , Arginina/metabolismo , Fenômenos Químicos , AMP Cíclico/metabolismo , Células HEK293 , Humanos , Masculino , Microssomos Hepáticos/efeitos dos fármacos , Microssomos Hepáticos/metabolismo , Ornitina/metabolismo , Limiar da Dor/efeitos dos fármacos , Peptidomiméticos/química , Ligação Proteica/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Receptores de Neuropeptídeos/antagonistas & inibidores , Receptores de Neuropeptídeos/metabolismo , Relação Estrutura-Atividade , Fatores de Tempo , Trítio/farmacocinéticaRESUMO
Mammalian RF-amide peptides are encoded by five different genes and act through five different G protein-coupled receptors. RF-amide-related peptides-1 and -3, neuropeptides AF and FF, Prolactin releasing peptides, Kisspeptins and RFa peptides are currently considered endogenous peptides for NPFF1, NPFF2, GPR10, GPR54 and GPR103 receptors, respectively. However, several studies suggest that the selectivity of these peptides for their receptors is low and indicate that expression patterns for receptors and their corresponding ligands only partially overlap. In this study, we took advantage of the cloning of the five human RF-amide receptors to systematically examine their affinity for and their activation by all human RF-amide peptides. Binding experiments, performed on membranes from CHO cells expressing GPR10, GPR54 and GPR103 receptors, confirmed their high affinity and remarkable selectivity for their cognate ligands. Conversely, NPFF1 and NPFF2 receptors displayed high affinity for all RF-amide peptides. Moreover, GTPγS and cAMP experiments showed that almost all RF-amide peptides efficiently activate NPFF1 and NPFF2 receptors. As NPFF is known to modulate morphine analgesia, we undertook a systematic analysis in mice of the hyperalgesic and anti morphine-induced analgesic effects of a representative set of endogenous RF-amide peptides. All of them induced hyperalgesia and/or prevented morphine analgesia following intracerebroventricular administration. Importantly, these effects were prevented by administration of RF9, a highly selective NPFF1/NPFF2 antagonist. Altogether, our results show that all endogenous RF-amide peptides display pain-modulating properties and point to NPFF receptors as essential players for these effects.
Assuntos
Analgésicos Opioides/farmacologia , Kisspeptinas/metabolismo , Morfina/farmacologia , Neuropeptídeos/metabolismo , Hormônio Liberador de Prolactina/metabolismo , Receptores de Neuropeptídeos/metabolismo , Animais , Células CHO , Cálcio/metabolismo , Cricetulus , AMP Cíclico/metabolismo , Guanosina 5'-O-(3-Tiotrifosfato)/farmacocinética , Humanos , Kisspeptinas/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neuropeptídeos/genética , Limiar da Dor/efeitos dos fármacos , Hormônio Liberador de Prolactina/genética , Ligação Proteica/efeitos dos fármacos , Fatores de Tempo , Trítio/farmacocinéticaRESUMO
We previously generated and characterized a genuine estrogen receptor (ER) ß-null mouse line (named ERß(ST)(L-/L-)) and showed that ERß(ST)(L-/L-) mice were sterile, due to an ovulation impairment in females and to an unknown reason in males, as their reproductive organs and spermatozoid motility appeared normal. We report here an assessment of the sexual behavior of ERß(ST)(L-/L-) null mice. We found that ERß(ST)(L-/L-) males display mildly impaired sexual behavior and that ERß(ST)(L-/L-) females are significantly less receptive and less attractive than wild-type (WT) females. Decreased attractivity is also exhibited by ERßAF2(0) but not by ERßAF1(0) mutant females (females devoid of either AF2 or AF1 activation function of ERß). Interestingly, by using an odor preference test, we have determined that the low attractiveness of ERß(ST)(L-/L-) and ERßAF2(0) females is related to a deficiency of a volatile chemosignal.