RESUMO
Recent studies reveal that airway epithelial cells are critical pulmonary circadian pacemaker cells, mediating rhythmic inflammatory responses. Using mouse models, we now identify the rhythmic circadian repressor REV-ERBα as essential to the mechanism coupling the pulmonary clock to innate immunity, involving both myeloid and bronchial epithelial cells in temporal gating and determining amplitude of response to inhaled endotoxin. Dual mutation of REV-ERBα and its paralog REV-ERBß in bronchial epithelia further augmented inflammatory responses and chemokine activation, but also initiated a basal inflammatory state, revealing a critical homeostatic role for REV-ERB proteins in the suppression of the endogenous proinflammatory mechanism in unchallenged cells. However, REV-ERBα plays the dominant role, as deletion of REV-ERBß alone had no impact on inflammatory responses. In turn, inflammatory challenges cause striking changes in stability and degradation of REV-ERBα protein, driven by SUMOylation and ubiquitination. We developed a novel selective oxazole-based inverse agonist of REV-ERB, which protects REV-ERBα protein from degradation, and used this to reveal how proinflammatory cytokines trigger rapid degradation of REV-ERBα in the elaboration of an inflammatory response. Thus, dynamic changes in stability of REV-ERBα protein couple the core clock to innate immunity.
Assuntos
Relógios Circadianos/imunologia , Ritmo Circadiano/imunologia , Homeostase/imunologia , Imunidade Inata , Membro 1 do Grupo D da Subfamília 1 de Receptores Nucleares/imunologia , Pneumonia/imunologia , Animais , Relógios Circadianos/genética , Ritmo Circadiano/genética , Homeostase/genética , Camundongos , Camundongos Transgênicos , Membro 1 do Grupo D da Subfamília 1 de Receptores Nucleares/genética , Pneumonia/genética , Pneumonia/patologia , Proteólise , Sumoilação/genética , Sumoilação/imunologiaRESUMO
The isomers of dibenzylamino-1-methylcyclohexan-1-ol and dibenzylamino-1-trifluoromethylcyclohexan-1-ol have been prepared. The stereochemistry of these compounds was unequivocally assigned through a combination of NMR spectroscopy and single crystal X-ray analysis. The cis-isomer of 3-N,N-dibenzylamino-1-trifluoromethylcyclohexanol and its derivatives display an unusual conformational behaviour in both solution-phase and the solid-state, where the amino group usually adopts an axial conformation.
Assuntos
Benzilaminas/síntese química , Cicloexanóis/síntese química , Benzilaminas/química , Cristalografia por Raios X , Cicloexanóis/química , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Conformação Molecular , EstereoisomerismoRESUMO
REV-ERBα has emerged as an important target for regulation of circadian rhythm and its associated physiology. Herein, we report on the optimization of a series of REV-ERBα agonists based on GSK4112 (1) for potency, selectivity, and bioavailability. (1) Potent REV-ERBα agonists 4, 10, 16, and 23 are detailed for their ability to suppress BMAL and IL-6 expression from human cells while also demonstrating excellent selectivity over LXRα. Amine 4 demonstrated in vivo bioavailability after either iv or oral dosing.
Assuntos
Aminas/síntese química , Membro 1 do Grupo D da Subfamília 1 de Receptores Nucleares/agonistas , Aminas/química , Aminas/farmacologia , Animais , Disponibilidade Biológica , Proteínas de Transporte/metabolismo , Linhagem Celular , Ritmo Circadiano , Glicina/análogos & derivados , Glicina/síntese química , Glicina/química , Glicina/farmacologia , Humanos , Receptores X do Fígado , Camundongos , Camundongos Endogâmicos C57BL , Receptores Nucleares Órfãos/metabolismo , Fragmentos de Peptídeos/metabolismo , Proteínas de Ligação a RNA , Ensaio Radioligante , Relação Estrutura-Atividade , Tiofenos/síntese química , Tiofenos/química , Tiofenos/farmacologiaRESUMO
The 2,3,4-trideoxy-2,3,4-trifluoro hexose analogues of d-glucose and d-altrose were prepared and characterised and these novel sugar analogues were explored by 2D-(19)F-EXSY NMR for their potential to cross erythrocyte (red blood cell) membranes, by comparison with the well known capacity of erythrocytes to transport d-glucose.
