RESUMEN
Cytokine storm during viral infection is a prospective predictor of morbidity and mortality, yet the cellular sources remain undefined. Here, using genetic and chemical tools to probe functions of the S1P(1) receptor, we elucidate cellular and signaling mechanisms that are important in initiating cytokine storm. Whereas S1P(1) receptor is expressed on endothelial cells and lymphocytes within lung tissue, S1P(1) agonism suppresses cytokines and innate immune cell recruitment in wild-type and lymphocyte-deficient mice, identifying endothelial cells as central regulators of cytokine storm. Furthermore, our data reveal immune cell infiltration and cytokine production as distinct events that are both orchestrated by endothelial cells. Moreover, we demonstrate that suppression of early innate immune responses through S1P(1) signaling results in reduced mortality during infection with a human pathogenic strain of influenza virus. Modulation of endothelium with a specific agonist suggests that diseases in which amplification of cytokine storm is a significant pathological component could be chemically tractable.
Asunto(s)
Citocinas/inmunología , Células Endoteliales/inmunología , Subtipo H1N1 del Virus de la Influenza A/fisiología , Gripe Humana/inmunología , Infecciones por Orthomyxoviridae/inmunología , Animales , Modelos Animales de Enfermedad , Humanos , Interferones/inmunología , Pulmón/citología , Pulmón/inmunología , Pulmón/virología , Linfocitos/inmunología , Ratones , Ratones Endogámicos C57BL , Infecciones por Orthomyxoviridae/patología , Infecciones por Orthomyxoviridae/virología , Receptores de Lisoesfingolípidos/agonistas , Transducción de SeñalRESUMEN
BACKGROUND: Mas-related G protein-coupled receptor X2 (MRGPRX2) is a promiscuous receptor on mast cells that mediates IgE-independent degranulation and has been implicated in multiple mast cell-mediated disorders, including chronic urticaria, atopic dermatitis, and pain disorders. Although it is a promising therapeutic target, few potent, selective, small molecule antagonists have been identified, and functional effects of human MRGPRX2 inhibition have not been evaluated in vivo. OBJECTIVE: We sought to identify and characterize novel, potent, and selective orally active small molecule MRGPRX2 antagonists for potential treatment of mast cell-mediated disease. METHODS: Antagonists were identified using multiple functional assays in cell lines overexpressing human MRGPRX2, LAD2 mast cells, human peripheral stem cell-derived mast cells, and isolated skin mast cells. Skin mast cell degranulation was evaluated in Mrgprb2em(-/-) knockout and Mrgprb2em(MRGPRX2) transgenic human MRGPRX2 knock-in mice by assessment of agonist-induced skin vascular permeability. Ex vivo skin mast cell degranulation and associated histamine release was evaluated by microdialysis of human skin tissue samples. RESULTS: MRGPRX2 antagonists potently inhibited agonist-induced MRGPRX2 activation and mast cell degranulation in all mast cell types tested in an IgE-independent manner. Orally administered MRGPRX2 antagonists also inhibited agonist-induced degranulation and resulting vascular permeability in MRGPRX2 knock-in mice. In addition, antagonist treatment dose dependently inhibited agonist-induced degranulation in ex vivo human skin. CONCLUSIONS: MRGPRX2 small molecule antagonists potently inhibited agonist-induced mast cell degranulation in vitro and in vivo as well as ex vivo in human skin, supporting potential therapeutic utility as a novel treatment for multiple human diseases involving clinically relevant mast cell activation.
Asunto(s)
Degranulación de la Célula , Mastocitos , Proteínas del Tejido Nervioso , Receptores Acoplados a Proteínas G , Receptores de Neuropéptido , Mastocitos/efectos de los fármacos , Mastocitos/inmunología , Animales , Receptores Acoplados a Proteínas G/antagonistas & inhibidores , Receptores Acoplados a Proteínas G/genética , Degranulación de la Célula/efectos de los fármacos , Humanos , Receptores de Neuropéptido/antagonistas & inhibidores , Receptores de Neuropéptido/genética , Ratones , Proteínas del Tejido Nervioso/genética , Ratones Noqueados , Piel/inmunología , Piel/efectos de los fármacos , Línea Celular , Ratones Endogámicos C57BLRESUMEN
Modification on a lead series of [1,4]oxazino[3,2-g]quinolin-7-ones at the 2-position led to selective androgen receptor modulators with improved in vivo activity. The most potent analog (-)-33a exhibited full maintenance of levator ani muscle at 3mg/kg and reduced activity on ventral prostate weight in a 2-week orally-dosed and orchidectomized rat maintenance assay.
Asunto(s)
Anabolizantes/farmacología , Oxazinas/farmacología , Próstata/efectos de los fármacos , Quinolonas/farmacología , Receptores Androgénicos , Administración Oral , Anabolizantes/síntesis química , Antagonistas de Receptores Androgénicos , Andrógenos , Animales , Masculino , Modelos Químicos , Orquiectomía , Tamaño de los Órganos/efectos de los fármacos , Oxazinas/síntesis química , Próstata/anatomía & histología , Quinolonas/síntesis química , Ratas , Ratas Sprague-Dawley , Relación Estructura-Actividad , Testosterona/farmacologíaRESUMEN
The androgen receptor is a ligand inducible transcription factor that is involved in a broad range of physiological functions. Here we describe the discovery of a new class of orally available selective androgen receptor modulators. The lead compound, 6-[(2R,5R)-2-methyl-5-((R)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl]-4-trifluoromethylquinolin-2(1H)-one (6a), showed excellent anabolic activity in muscle with reduced effect on the prostate in a rat model of hypogonadism. The compound also improved bone strength in a rat model of post-menopausal osteoporosis.
Asunto(s)
Anabolizantes/síntesis química , Antagonistas de Receptores Androgénicos , Andrógenos , Conservadores de la Densidad Ósea/síntesis química , Pirrolidinas/síntesis química , Quinolinas/síntesis química , Quinolonas/síntesis química , Administración Oral , Anabolizantes/farmacocinética , Anabolizantes/farmacología , Animales , Disponibilidad Biológica , Conservadores de la Densidad Ósea/farmacocinética , Conservadores de la Densidad Ósea/farmacología , Femenino , Humanos , Hipogonadismo/tratamiento farmacológico , Hipogonadismo/patología , Masculino , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/metabolismo , Tamaño de los Órganos/efectos de los fármacos , Osteoporosis Posmenopáusica/tratamiento farmacológico , Próstata/efectos de los fármacos , Pirrolidinas/farmacocinética , Pirrolidinas/farmacología , Quinolinas/farmacocinética , Quinolinas/farmacología , Quinolonas/farmacocinética , Quinolonas/farmacología , Ratas , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
A series of selective androgen receptor modulators (SARMs) with a wide spectrum of receptor modulating activities was developed based on optimization of the 4-substituted 6-bisalkylamino-2-quinolinones (3). Significance of the trifluoromethyl group on the side chains and its interactions with amino acid residues within the androgen receptor (AR) ligand binding domain are discussed. A representative analog (9) was tested orally in a rodent model of hypogonadism and demonstrated desirable tissue selectivity.
Asunto(s)
Quinolinas/química , Quinolinas/síntesis química , Receptores Androgénicos/efectos de los fármacos , Antagonistas de Andrógenos/síntesis química , Antagonistas de Andrógenos/farmacología , Antagonistas de Receptores Androgénicos , Andrógenos , Animales , Unión Competitiva/efectos de los fármacos , Dihidrotestosterona/antagonistas & inhibidores , Dihidrotestosterona/farmacología , Genitales Masculinos/efectos de los fármacos , Hipogonadismo/tratamiento farmacológico , Indicadores y Reactivos , Espectroscopía de Resonancia Magnética , Masculino , Modelos Moleculares , Conformación Molecular , Orquiectomía , Tamaño de los Órganos/efectos de los fármacos , Especificidad de Órganos , Próstata/efectos de los fármacos , Ratas , Relación Estructura-Actividad , TransfecciónRESUMEN
A series of 5-benzylidene-1,2-dihydro-2,2,4-trimethyl-5H-1-aza-6-oxa-chrysenes was synthesized and profiled for their ability to act as selective glucocorticoid receptor modulators (SGRMs). The synthesis and structure-activity relationships for this series of compounds are presented.