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Victoria J Barrett - One of the best experts on this subject based on the ideXlab platform.
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pharmacological characterization of gsk573719 umeclidinium a novel long acting inhaled antagonist of the muscarinic Cholinergic Receptors for treatment of pulmonary diseases
Journal of Pharmacology and Experimental Therapeutics, 2013Co-Authors: Michael Salmon, James J Foley, Peter T Buckley, Dulcie B Schmidt, Miriam Burman, Edward F Webb, Christopher J Dehaas, Charles J Kotzer, Mark A. Luttmann, Victoria J BarrettAbstract:Activation of M3 muscarinic Cholinergic Receptors (mAChR) increases airway tone whereas its blockade improves lung function and quality of life in patients with pulmonary diseases. The present study evaluated the pharmacological properties of a novel mAChR antagonist, GSK573719, 4-[hydroxy(diphenyl)methyl]-1-{2-[(phenylmethyl)oxy]ethyl}-1-azoniabicyclo[2.2.2]octane (umeclidinium). The affinity (K i ) of GSK573719 for the cloned human M1-M5 mAChRs ranged from 0.05-0.16 nM. Dissociation of [ 3 H]-GSK573719 from the M3 mAChR was slower than that for the M2 mAChR; t 1/2 values = 82 and 9 min, respectively. In CHO cells transfected with recombinant human M3 mAChRs, GSK573719 demonstrated pM potency (–log pA 2 = 23.9 pM) in an acetylcholine (Ach)-mediated Ca 2+ mobilization assay. Concentration-response curves indicate competitive antagonism with partial reversibility after drug wash-out. Using isolated human bronchial strips, GSK573719 was also potent and showed competitive antagonism (–log pA 2 = 316 pM) vs carbachol and was slowly reversible in a concentration-dependent manner (1–100 nM). The time to 50% restoration of contraction at 10 nM was about 381 min (vs 413 min for tiotropium bromide). In mice, the ED 50 value was 0.02 μg/mouse intranasally. In conscious guinea pigs, intratracheal administration of GSK573719 dose-dependently blocked Ach-induced bronchoconstriction with long duration of action and was comparable to tiotropium; 2.5 μg elicited 50% bronchoprotection for >24h. Thus, GSK573719 is a potent antiCholinergic agent that demonstrates slow functional reversibility at the human M3 mAChR and long duration of action in animal models. This pharmacological profile translated into 24h duration of bronchodilation in vivo which suggested umeclidinium will be a once-daily inhaled treatment for pulmonary diseases.
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pharmacological characterization of gsk573719 umeclidinium a novel long acting inhaled antagonist of the muscarinic Cholinergic Receptors for treatment of pulmonary diseases
Journal of Pharmacology and Experimental Therapeutics, 2013Co-Authors: Michael Salmon, James J Foley, Peter T Buckley, Dulcie B Schmidt, Miriam Burman, Edward F Webb, Christopher J Dehaas, Charles J Kotzer, Mark A. Luttmann, Victoria J BarrettAbstract:Activation of muscarinic subtype 3 (M3) muscarinic Cholinergic Receptors (mAChRs) increases airway tone, whereas its blockade improves lung function and quality of life in patients with pulmonary diseases. The present study evaluated the pharmacological properties of a novel mAChR antagonist, GSK573719 (4-[hydroxy(diphenyl)methyl]-1-{2-[(phenylmethyl)oxy]ethyl}-1-azoniabicyclo[2.2.2]octane; umeclidinium). The affinity (Ki) of GSK573719 for the cloned human M1-M5 mAChRs ranged from 0.05 to 0.16 nM. Dissociation of [(3)H]GSK573719 from the M3 mAChR was slower than that for the M2 mAChR [half-life (t1/2) values: 82 and 9 minutes, respectively]. In Chinese hamster ovary cells transfected with recombinant human M3 mAChRs, GSK573719 demonstrated picomolar potency (-log pA2 = 23.9 pM) in an acetylcholine (Ach)-mediated Ca(2+) mobilization assay. Concentration-response curves indicate competitive antagonism with partial reversibility after drug washout. Using isolated human bronchial strips, GSK573719 was also potent and showed competitive antagonism (-log pA2 = 316 pM) versus carbachol, and was slowly reversible in a concentration-dependent manner (1-100 nM). The time to 50% restoration of contraction at 10 nM was about 381 minutes (versus 413 minutes for tiotropium bromide). In mice, the ED50 value was 0.02 μg/mouse intranasally. In conscious guinea pigs, intratracheal administration of GSK573719 dose dependently blocked Ach-induced bronchoconstriction with long duration of action, and was comparable to tiotropium; 2.5 μg elicited 50% bronchoprotection for >24 hours. Thus, GSK573719 is a potent antiCholinergic agent that demonstrates slow functional reversibility at the human M3 mAChR and long duration of action in animal models. This pharmacological profile translated into a 24-hour duration of bronchodilation in vivo, which suggested umeclidinium will be a once-daily inhaled treatment of pulmonary diseases.
Sergei A. Grando - One of the best experts on this subject based on the ideXlab platform.
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plasticity of the murine spleen t cell Cholinergic Receptors and their role in in vitro differentiation of naive cd4 t cells toward the th1 th2 and th17 lineages
Genes and Immunity, 2011Co-Authors: Jing Qian, Valentin Galitovskiy, Alexander I Chernyavsky, Steve Marchenko, Sergei A. GrandoAbstract:Acetylcholine (ACh) regulates vital functions of T cells by acting on the nicotinic and muscarinic classes of Cholinergic Receptors, nAChR and mAChRs, respectively. This study was performed in murine splenic T cells. In freshly isolated CD4 and CD8 T cells, we detected mRNAs encoding α5, α9, α10, β1, β2, β4 nAChR subunits and M₁, M₃, M₄ and M₅ mAChR subtypes, whereas α2 was detected only in CD8 T cells. In vitro activation of CD4 T cells through T-cell receptor (TCR)/CD3 cross-linking was associated with the appearance of α4 and α7, upregulation of α5, α10, β4, M₁ and M₅ and downregulation of α9 and β2, whereas in vitro activation of CD8 T cells also featured the appearance of α4 and α7, as well as upregulation of α2, α5, β4, M₁ and M₄, and downregulation of α10, β1, β2 and M₃. In vitro polarization toward T helper (Th) 1 lineage was associated with a decrease of β2, β4 and M₃ expression; that toward Th2 cells with downregulation of α9 and M₃, and upregulation of M₁ and M₅; and that toward Th17 phenotype with downregulation of α9, α10, β2 and M₃ mAChR. Polarized T cells also expressed α4, but not α1, α2, α3, α6, β3 or M₂. To determine the role of Cholinergic Receptors in mediating the immunoregulatory action of autocrine/paracrine ACh, we analyzed the effects of nicotinic and muscarinic agonists±antagonists on cytokine production in the CD4+CD62L+ T cells co-stimulated via TCR/CD3 cross-linking. The nicotinergic stimulation upregulated interferon-γ (IFN-γ) and downregulated interleukin (IL)-17 secretion, whereas the muscarinic stimulation enhanced IL-10 and IL-17 and inhibited INF-γ secretion. These results demonstrated plasticity of the T-cell Cholinergic system.
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the pathophysiological significance of nondesmoglein targets of pemphigus autoimmunity development of antibodies against keratinocyte Cholinergic Receptors in patients with pemphigus vulgaris and pemphigus foliaceus
Archives of Dermatology, 1998Co-Authors: Vu Thuong Nguyen, Tou X Lee, Assane Ndoye, Leonard D Shultz, Mark R Pittelkow, Mark V Dahl, Peter J Lynch, Sergei A. GrandoAbstract:Objectives To determine whether nondesmoglein (non-Dsg) autoantibodies are pathogenic and whether they recognize keratinocyte Cholinergic Receptors that control cell adhesion because antikeratinocyte autoimmunity in patients with pemphigus vulgaris is not limited to the development of autoantibodies to Dsg. Design To determine whether non-Dsg autoantibodies are pathogenic, we sought to induce pemphigus in genetically engineered neonatal mice lacking Dsg 3 using pemphigus vulgaris IgGs that did not cross-react with Dsg 1. To determine whether pemphigus autoimmunity involves keratinocyte Cholinergic Receptors, the latter were separated from cell membranes of human keratinocytes, tagged with the covalent label [ 3 H]propylbenzilylcholine mustard, and used as an antigen in a radioimmunoprecipitation assay of 34 pemphigus vulgaris and 6 pemphigus foliaceus serum samples. Setting The dermatologic clinics of the University of Minnesota, Minneapolis; the Mayo Clinic, Rochester, Minn; and the University of California–Davis Medical Center, Sacramento. Patients Serum samples were collected from 34 patients with pemphigus vulgaris and 6 patients with pemphigus foliaceus (aged 31-89 years) and from 7 age-similar patients of both sexes with nonpemphigus blistering or the following immune-mediated conditions: pemphigoid gestationis, bullous drug eruption, lupus erythematosus, erythema nodosum, urticaria, acute contact dermatitis, and skin ulcers. Main Outcome Measures Clinical, laboratory, and histopathologic findings. Results Extensive skin blistering accompanied by the Nikolsky sign and suprabasilar acantholysis was induced in the Dsg3null mice that received pemphigus, but not normal human IgGs. In the radioimmunoprecipitation assays for reactivity with Cholinergic Receptors, the mean radioactivity precipitated by pemphigus serum samples significantly exceeded both normal- and disease-control levels ( P =.001-.02). The mean individual levels of radioactivity precipitated by 34 pemphigus vulgaris and pemphigus foliaceus serum samples (85%) exceeded control values by a mean of approximately 2.6 times. Conclusions Autoantibodies to keratinocyte cell-surface molecules other than Dsg 1 and Dsg 3 can induce clinical features of pemphigus vulgaris. Patients with pemphigus vulgaris and those with pemphigus foliaceus develop IgG antibodies that precipitate radiolabeled Cholinergic Receptors. Because these Receptors control keratinocyte adhesion and motility, their inactivation by autoantibodies may elicit intracellular signals that cause disassembly of desmosomes, leading to acantholysis and blistering.
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Biological Functions of Keratinocyte Cholinergic Receptors
The journal of investigative dermatology. Symposium proceedings, 1997Co-Authors: Sergei A. GrandoAbstract:Human epidermal keratinocytes possess Cholinergic enzymes, which synthesize and degrade acetylcholine, and express both nicotinic and muscarinic classes of Cholinergic Receptors on their cell surfaces. These Receptors bind acetylcholine and initiate cellular response. The presence in keratinocytes of a functional Cholinergic system suggests a role for acetylcholine in most, if not all, aspects of keratinocyte function. Autocrine and paracrine acetylcholine are required to sustain the viability of keratinocytes in vitro, and Cholinergic drugs can alter keratinocyte proliferation, adhesion, migration, and differentiation. Acetylcholine employs calcium as a mediator for its effects on keratinocytes. In turn, changes in calcium concentration may affect expression and function of keratinocyte Cholinergic enzymes and Cholinergic Receptors. At different stages of their differentiation, keratinocytes may demonstrate unique combinations of Cholinergic enzymes and Cholinergic receptor types. This would allow basal, prickle, and granular keratinocytes to respond to acetylcholine differently, in accordance with their functions at each stage of keratinocyte development in epidermis. Journal of Investigative Dermatology Symposium Proceedings 2:41–48, 1997
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activation of keratinocyte nicotinic Cholinergic Receptors stimulates calcium influx and enhances cell differentiation
Journal of Investigative Dermatology, 1996Co-Authors: Sergei A. Grando, Tou X Lee, Robert M Horton, Theodora M Mauro, David A Kist, Mark V DahlAbstract:Human epidermal keratinocytes synthesize, secrete, and degrade acetylcholine and use their cell-surface nicotinic and muscarinic Cholinergic Receptors to mediate the autocrine and paracrine effects of acetylcholine. Because acetylcholine modulates transmembrane Ca 2+ transport and intracellular metabolism in several types of cells, we hypothesized that Cholinergic agents might have similar effects on keratinocytes. Nicotine increased in a concentration-dependent manner the amount of 45 Ca 2+ taken up by keratinocytes isolated from human neonatal foreskins. This effect was abolished in the presence of the specific nicotinic antagonist mecamylamine, indicating that it was mediated by keratinocyte nicotinic acetylcholine receptor(s). The sequences encoding the α 5 and α 7 nicotinic receptor subunits were amplified from cDNA isolated from cultured keratinocytes. These subunits, as well as the α 3, β 2, and β 4 subunits previously found in keratinocytes, can be components of Ca 2+ -permeable nicotinic receptor channels. To learn how activation of keratinocyte nicotinic Receptors affected the rate of cell differentiation, we measured the nicotinic Cholinergic effects on the expression of differentiation markers by cultured keratinocytes. Long-term incubations with micromolar concentrations of nicotine markedly increased the number of cells forming cornified envelopes and the number of cells staining with antibodies to suprabasal keratin 10, transglutaminase type I, involucrin, and filaggrin. Because nicotinic Cholinergic stimulation causes transmembrane Ca 2+ transport into keratinocytes, and because changes in concentrations of intracellular Ca 2+ are known to alter various keratinocyte functions, including differentiation, the subcellular mechanisms mediating the autocrine and paracrine actions of epidermal acetylocholine on keratinocytes may involve Ca 2+ as a second messenger.
Allan C. Collins - One of the best experts on this subject based on the ideXlab platform.
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genetic correlation of inhibitory gating of hippocampal auditory evoked response and α bungarotoxin binding nicotinic Cholinergic Receptors in inbred mouse strains
Neuropsychopharmacology, 1996Co-Authors: Michael J. Marks, Allan C. Collins, Karen E Stevens, Robert Freedman, Michael E Hall, Sherry Leonard, Gregory M RoseAbstract:One function of the hippocampus is to ascertain the novelty of incoming sensations and encode significant new information into memory. The regulation of response to repeated stimuli may prevent overloading of this function by redundant sensory input. Recent pharmacological studies implicate the role of α-bungarotoxin-sensitive nicotinic Cholinergic Receptors in the inhibition of hippocampal response to repeated auditory stimuli. The number of hippocampal α-bungarotoxin-sensitive Receptors has a major genetic determinant, as demonstrated by a significant variance between different inbred mouse strains. The purpose of the present study was to determine whether there was a related genetic correlation for the gating of auditory response. Nine inbred mouse strains, representing a continuum of hippocampal α-bungarotoxin binding, were tested for the electrophysiological response to repeated auditory stimulation, followed by whole hippocampus membrane α-bungarotoxin binding studies. Several parameters of the auditory evoked response showed significant genetic variance over the nine strains, and a significant correlation was found between hippocampal α-bungarotoxin binding and both the amplitude of the initial evoked response and its inhibition to repeated auditory stimuli. There was no correlation of the auditory evoked response with high-affinity nicotine binding. These data further support the hypothesis that α-bungarotoxin-sensitive nicotinic Receptors are involved in the regulation of hippocampal response to repeated auditory stimuli and suggest that this function is genetically controlled.
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An autoradiographic analysis of Cholinergic Receptors in mouse brain after chronic nicotine treatment.
The Journal of pharmacology and experimental therapeutics, 1991Co-Authors: James R. Pauly, Michael J. Marks, S D Gross, Allan C. CollinsAbstract:Quantitative autoradiographic procedures were used to examine the effects of chronic nicotine infusion on the number of central nervous system nicotinic Cholinergic Receptors. Female DBA mice were implanted with jugular cannulas and infused with saline or various doses of nicotine (0.25, 0.5, 1.0 or 2.0 mg/kg/hr) for 10 days. The animals were then sacrificed and the brains were removed and frozen in isopentane. Cryostat sections were collected and prepared for autoradiographic procedures as previously described. Nicotinic Cholinergic Receptors were labeled with L-[3H]nicotine or alpha-[125I]bungarotoxin; [3H]quinuclidinyl benzilate was used to measure muscarinic Cholinergic receptor binding. Chronic nicotine infusion increased the number of sites labeled by [3H]nicotine in most brain areas. However, the extent of the increase in binding as well as the dose-response curves for the increase were widely different among brain regions. After the highest treatment dose, binding was increased in 67 of 86 regions measured. Septal and thalamic regions were most resistant to change. Nicotinic binding measured by alpha-[125I]bungarotoxin also increased after chronic treatment, but in a less robust fashion. At the highest treatment dose, only 26 of 80 regions were significantly changes. Muscarinic binding was not altered after chronic nicotine treatment. These data suggest that brain regions are not equivalent in the mechanisms that regulate alterations in nicotinic Cholinergic receptor binding after chronic nicotine treatment.
Michael Salmon - One of the best experts on this subject based on the ideXlab platform.
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pharmacological characterization of gsk573719 umeclidinium a novel long acting inhaled antagonist of the muscarinic Cholinergic Receptors for treatment of pulmonary diseases
Journal of Pharmacology and Experimental Therapeutics, 2013Co-Authors: Michael Salmon, James J Foley, Peter T Buckley, Dulcie B Schmidt, Miriam Burman, Edward F Webb, Christopher J Dehaas, Charles J Kotzer, Mark A. Luttmann, Victoria J BarrettAbstract:Activation of M3 muscarinic Cholinergic Receptors (mAChR) increases airway tone whereas its blockade improves lung function and quality of life in patients with pulmonary diseases. The present study evaluated the pharmacological properties of a novel mAChR antagonist, GSK573719, 4-[hydroxy(diphenyl)methyl]-1-{2-[(phenylmethyl)oxy]ethyl}-1-azoniabicyclo[2.2.2]octane (umeclidinium). The affinity (K i ) of GSK573719 for the cloned human M1-M5 mAChRs ranged from 0.05-0.16 nM. Dissociation of [ 3 H]-GSK573719 from the M3 mAChR was slower than that for the M2 mAChR; t 1/2 values = 82 and 9 min, respectively. In CHO cells transfected with recombinant human M3 mAChRs, GSK573719 demonstrated pM potency (–log pA 2 = 23.9 pM) in an acetylcholine (Ach)-mediated Ca 2+ mobilization assay. Concentration-response curves indicate competitive antagonism with partial reversibility after drug wash-out. Using isolated human bronchial strips, GSK573719 was also potent and showed competitive antagonism (–log pA 2 = 316 pM) vs carbachol and was slowly reversible in a concentration-dependent manner (1–100 nM). The time to 50% restoration of contraction at 10 nM was about 381 min (vs 413 min for tiotropium bromide). In mice, the ED 50 value was 0.02 μg/mouse intranasally. In conscious guinea pigs, intratracheal administration of GSK573719 dose-dependently blocked Ach-induced bronchoconstriction with long duration of action and was comparable to tiotropium; 2.5 μg elicited 50% bronchoprotection for >24h. Thus, GSK573719 is a potent antiCholinergic agent that demonstrates slow functional reversibility at the human M3 mAChR and long duration of action in animal models. This pharmacological profile translated into 24h duration of bronchodilation in vivo which suggested umeclidinium will be a once-daily inhaled treatment for pulmonary diseases.
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pharmacological characterization of gsk573719 umeclidinium a novel long acting inhaled antagonist of the muscarinic Cholinergic Receptors for treatment of pulmonary diseases
Journal of Pharmacology and Experimental Therapeutics, 2013Co-Authors: Michael Salmon, James J Foley, Peter T Buckley, Dulcie B Schmidt, Miriam Burman, Edward F Webb, Christopher J Dehaas, Charles J Kotzer, Mark A. Luttmann, Victoria J BarrettAbstract:Activation of muscarinic subtype 3 (M3) muscarinic Cholinergic Receptors (mAChRs) increases airway tone, whereas its blockade improves lung function and quality of life in patients with pulmonary diseases. The present study evaluated the pharmacological properties of a novel mAChR antagonist, GSK573719 (4-[hydroxy(diphenyl)methyl]-1-{2-[(phenylmethyl)oxy]ethyl}-1-azoniabicyclo[2.2.2]octane; umeclidinium). The affinity (Ki) of GSK573719 for the cloned human M1-M5 mAChRs ranged from 0.05 to 0.16 nM. Dissociation of [(3)H]GSK573719 from the M3 mAChR was slower than that for the M2 mAChR [half-life (t1/2) values: 82 and 9 minutes, respectively]. In Chinese hamster ovary cells transfected with recombinant human M3 mAChRs, GSK573719 demonstrated picomolar potency (-log pA2 = 23.9 pM) in an acetylcholine (Ach)-mediated Ca(2+) mobilization assay. Concentration-response curves indicate competitive antagonism with partial reversibility after drug washout. Using isolated human bronchial strips, GSK573719 was also potent and showed competitive antagonism (-log pA2 = 316 pM) versus carbachol, and was slowly reversible in a concentration-dependent manner (1-100 nM). The time to 50% restoration of contraction at 10 nM was about 381 minutes (versus 413 minutes for tiotropium bromide). In mice, the ED50 value was 0.02 μg/mouse intranasally. In conscious guinea pigs, intratracheal administration of GSK573719 dose dependently blocked Ach-induced bronchoconstriction with long duration of action, and was comparable to tiotropium; 2.5 μg elicited 50% bronchoprotection for >24 hours. Thus, GSK573719 is a potent antiCholinergic agent that demonstrates slow functional reversibility at the human M3 mAChR and long duration of action in animal models. This pharmacological profile translated into a 24-hour duration of bronchodilation in vivo, which suggested umeclidinium will be a once-daily inhaled treatment of pulmonary diseases.
J. James Frost - One of the best experts on this subject based on the ideXlab platform.
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In vivo labeling of nicotinic Cholinergic Receptors in brain with [3H]cytisine.
Life Sciences, 2002Co-Authors: John E. Flesher, Ursula Scheffel, Edythe D. London, J. James FrostAbstract:Abstract [ 3 H]Cytisine was evaluated as an in vivo ligand for the nicotinic Cholinergic receptor (nAchR) in mouse brain. The tracer was injected intravenously, and radioactivity in brain regions was analyzed. Radioactivity peaked in the brain at 30 minutes. It was highest in the thalamus, intermediate in the superior colliculi, prefrontal cortex and hippocampus, and low in the cerebellum. Pretreatment with unlabeled cytisine inhibited binding in the thalamus, but not in the cerebellum. Binding was displaced by l-nicotine, but not by d-nicotine or dexetimide. The results suggest that cytisine, appropriately labeled with a positron emitting radionuclide, may be useful for study of nicotinic Cholinergic Receptors in humans by emission computed tomography.