Felder CC


Full name : Felder Christian C

First name : Christian C

Mail : Lilly Research Labs Drop 0510, Lilly Corporate Center, Indianapolis, IN 46285

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City :

Country : USA

Email : felder@lilly.com

Phone : I 317 276 1380

Fax : 1 317 276 7600

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References (49)

Title : M1 muscarinic allosteric modulators slow prion neurodegeneration and restore memory loss - Bradley_2017_J.Clin.Invest_127_487
Author(s) : Bradley SJ , Bourgognon JM , Sanger HE , Verity N , Mogg AJ , White DJ , Butcher AJ , Moreno JA , Molloy C , Macedo-Hatch T , Edwards JM , Wess J , Pawlak R , Read DJ , Sexton PM , Broad LM , Steinert JR , Mallucci GR , Christopoulos A , Felder CC , Tobin AB
Ref : J Clinical Investigation , 127 :487 , 2017
Abstract : The current frontline symptomatic treatment for Alzheimer's disease (AD) is whole-body upregulation of cholinergic transmission via inhibition of acetylcholinesterase. This approach leads to profound dose-related adverse effects. An alternative strategy is to selectively target muscarinic acetylcholine receptors, particularly the M1 muscarinic acetylcholine receptor (M1 mAChR), which was previously shown to have procognitive activity. However, developing M1 mAChR-selective orthosteric ligands has proven challenging. Here, we have shown that mouse prion disease shows many of the hallmarks of human AD, including progressive terminal neurodegeneration and memory deficits due to a disruption of hippocampal cholinergic innervation. The fact that we also show that muscarinic signaling is maintained in both AD and mouse prion disease points to the latter as an excellent model for testing the efficacy of muscarinic pharmacological entities. The memory deficits we observed in mouse prion disease were completely restored by treatment with benzyl quinolone carboxylic acid (BQCA) and benzoquinazoline-12 (BQZ-12), two highly selective positive allosteric modulators (PAMs) of M1 mAChRs. Furthermore, prolonged exposure to BQCA markedly extended the lifespan of diseased mice. Thus, enhancing hippocampal muscarinic signaling using M1 mAChR PAMs restored memory loss and slowed the progression of mouse prion disease, indicating that this ligand type may have clinical benefit in diseases showing defective cholinergic transmission, such as AD.
ESTHER : Bradley_2017_J.Clin.Invest_127_487
PubMedSearch : Bradley_2017_J.Clin.Invest_127_487
PubMedID: 27991860

Title : Characterization of PCS1055, a novel muscarinic M4 receptor antagonist - Croy_2016_Eur.J.Pharmacol_782_70
Author(s) : Croy CH , Chan WY , Castetter AM , Watt ML , Quets AT , Felder CC
Ref : European Journal of Pharmacology , 782 :70 , 2016
Abstract : Identification of synthetic ligands selective for muscarinic receptor subtypes has been challenging due to the high sequence identity and structural homology among the five muscarinic acetylcholine receptors. Here, we report the pharmacological characterization of PCS1055, a novel muscarinic M4 receptor antagonist. PCS1055 inhibited radioligand [(3)H]-NMS binding to the M4 receptor with a Ki=6.5nM. Though the potency of PCS1055 is lower than that of pan-muscarinic antagonist atropine, it has better subtype selectivity over previously reported M4-selective reagents such as the muscarinic-peptide toxins (Karlsson et al., 1994; Santiago and Potter, 2001a) at the M1 subtype, and benzoxazine ligand PD102807 at the M3-subtype (Bohme et al., 2002). A detailed head-to-head comparison study using [(3)H]-NMS competitive binding assays characterizes the selectivity profiles of PCS1055 to that of other potent muscarinic-antagonist compounds PD102807, tropicamide, AF-DX-384, pirenzapine, and atropine. In addition to binding studies, the subtype specificity of PCS1055 is also demonstrated by functional receptor activation as readout by GTP-gamma-[(35)S] binding. These GTP-gamma-[(35)S] binding studies showed that PCS1055 exhibited 255-, 69.1-, 342- and >1000-fold greater inhibition of Oxo-M activity at the M4 versus the M1-, M2(-), M3-or M5 receptor subtypes, respectively. Schild analyses indicates that PCS1055 acts as a competitive antagonist to muscarinic M4 receptor, and confirms the affinity of the ligand to be low nanomolar, Kb=5.72nM. Therefore, PCS1055 represents a new M4-preferring antagonist that may be useful in elucidating the roles of M4 receptor signaling.
ESTHER : Croy_2016_Eur.J.Pharmacol_782_70
PubMedSearch : Croy_2016_Eur.J.Pharmacol_782_70
PubMedID: 27085897

Title : M1 and m2 muscarinic receptor subtypes regulate antidepressant-like effects of the rapidly acting antidepressant scopolamine - Witkin_2014_J.Pharmacol.Exp.Ther_351_448
Author(s) : Witkin JM , Overshiner C , Li X , Catlow JT , Wishart GN , Schober DA , Heinz BA , Nikolayev A , Tolstikov VV , Anderson WH , Higgs RE , Kuo MS , Felder CC
Ref : Journal of Pharmacology & Experimental Therapeutics , 351 :448 , 2014
Abstract : Scopolamine produces rapid and significant symptom improvement in patients with depression, and most notably in patients who do not respond to current antidepressant treatments. Scopolamine is a nonselective muscarinic acetylcholine receptor antagonist, and it is not known which one or more of the five receptor subtypes in the muscarinic family are mediating these therapeutic effects. We used the mouse forced-swim test, an antidepressant detecting assay, in wild-type and transgenic mice in which each muscarinic receptor subtype had been genetically deleted to define the relevant receptor subtypes. Only the M1 and M2 knockout (KO) mice had a blunted response to scopolamine in the forced-swim assay. In contrast, the effects of the tricyclic antidepressant imipramine were not significantly altered by gene deletion of any of the five muscarinic receptors. The muscarinic antagonists biperiden, pirenzepine, and VU0255035 (N-[3-oxo-3-[4-(4-pyridinyl)-1-piper azinyl]propyl]-2,1,3-benzothiadiazole-4-sulfonamide) with selectivity for M1 over M2 receptors also demonstrated activity in the forced-swim test, which was attenuated in M1 but not M2 receptor KO mice. An antagonist with selectivity of M2 over M1 receptors (SCH226206 [(2-amino-3-methyl-phenyl)-[4-[4-[[4-(3 chlorophenyl)sulfonylphenyl]methyl]-1-piperidyl]-1-piperidyl]methanone]) was also active in the forced-swim assay, and the effects were deleted in M2 (-/-) mice. Brain exposure and locomotor activity in the KO mice demonstrated that these behavioral effects of scopolamine are pharmacodynamic in nature. These data establish muscarinic M1 and M2 receptors as sufficient to generate behavioral effects consistent with an antidepressant phenotype and therefore as potential targets in the antidepressant effects of scopolamine.
ESTHER : Witkin_2014_J.Pharmacol.Exp.Ther_351_448
PubMedSearch : Witkin_2014_J.Pharmacol.Exp.Ther_351_448
PubMedID: 25187432

Title : Activation and allosteric modulation of a muscarinic acetylcholine receptor - Kruse_2013_Nature_504_101
Author(s) : Kruse AC , Ring AM , Manglik A , Hu J , Hu K , Eitel K , Hubner H , Pardon E , Valant C , Sexton PM , Christopoulos A , Felder CC , Gmeiner P , Steyaert J , Weis WI , Garcia KC , Wess J , Kobilka BK
Ref : Nature , 504 :101 , 2013
Abstract : Despite recent advances in crystallography and the availability of G-protein-coupled receptor (GPCR) structures, little is known about the mechanism of their activation process, as only the beta2 adrenergic receptor (beta2AR) and rhodopsin have been crystallized in fully active conformations. Here we report the structure of an agonist-bound, active state of the human M2 muscarinic acetylcholine receptor stabilized by a G-protein mimetic camelid antibody fragment isolated by conformational selection using yeast surface display. In addition to the expected changes in the intracellular surface, the structure reveals larger conformational changes in the extracellular region and orthosteric binding site than observed in the active states of the beta2AR and rhodopsin. We also report the structure of the M2 receptor simultaneously bound to the orthosteric agonist iperoxo and the positive allosteric modulator LY2119620. This structure reveals that LY2119620 recognizes a largely pre-formed binding site in the extracellular vestibule of the iperoxo-bound receptor, inducing a slight contraction of this outer binding pocket. These structures offer important insights into the activation mechanism and allosteric modulation of muscarinic receptors.
ESTHER : Kruse_2013_Nature_504_101
PubMedSearch : Kruse_2013_Nature_504_101
PubMedID: 24256733

Title : Probe dependence in the allosteric modulation of a G protein-coupled receptor: implications for detection and validation of allosteric ligand effects - Valant_2012_Mol.Pharmacol_81_41
Author(s) : Valant C , Felder CC , Sexton PM , Christopoulos A
Ref : Molecular Pharmacology , 81 :41 , 2012
Abstract : We recently described 3-amino-5-chloro-6-methoxy-4-methylthieno[2,3-b]pyridine-2-carboxylic acid cyclopropylamide (LY2033298) as a novel allosteric modulator of M(4) muscarinic acetylcholine (ACh) receptors (mAChRs) on the basis of its ability to preferentially potentiate the actions of ACh at the M(4) mAChR subtype. In the current study, we show that LY2033298 can also bind to the M(2) mAChR and mediate robust positive or negative allosteric effects, depending on the orthosteric ligand used as a probe of receptor activity. This finding of striking "probe dependence" indicates that the previously described selectivity of the modulator does not arise as a consequence of selective affinity for a poorly conserved allosteric site but rather is due to subtype-selective cooperativity with ACh upon interaction with a common allosteric binding site. Moreover, a comparison of the effects of the modulator on orthosteric ligand affinity relative to signaling through a [(35)S]guanosine 5'-O-(3-thio)triphosphate or extracellular signal-regulated kinase 1/2 phosphorylation assay at the M(2) mAChR revealed that, although the effects on binding were positive in all instances, the effects on signaling were either positive or strongly negative, depending on the agonist and the pathway. Mutational analysis identified residues Tyr177 and Trp99(3.28) (Ballesteros and Weinstein numbers are provided in superscript to indicate relative position of residues within the transmembrane domain) as contributing to the binding of LY2033298, whereas the orthosteric site residues, Tyr104(3.33) and Tyr403(6.51), contributed to the ability of the ligand to impose pathway-biased modulation. Taken together, these findings have important implications for the detection and validation of allosteric modulators of G protein-coupled receptors (GPCRs), because they highlight the potential for ligand misclassification or lack of appreciation of off-target allosteric activities.
ESTHER : Valant_2012_Mol.Pharmacol_81_41
PubMedSearch : Valant_2012_Mol.Pharmacol_81_41
PubMedID: 21989256

Title : Impact of species variability and 'probe-dependence' on the detection and in vivo validation of allosteric modulation at the M4 muscarinic acetylcholine receptor - Suratman_2011_Br.J.Pharmacol_162_1659
Author(s) : Suratman S , Leach K , Sexton P , Felder CC , Loiacono R , Christopoulos A
Ref : British Journal of Pharmacology , 162 :1659 , 2011
Abstract : BACKGROUND AND PURPOSE: We recently characterized LY2033298 as a novel allosteric modulator and agonist at M(4) muscarinic acetylcholine receptors (mAChRs). Evidence also suggested a difference in the potency of LY2033298 at rodent relative to human M(4) mAChRs. The current study investigated the basis for the species difference of this modulator and used this knowledge to rationalize its in vivo actions. EXPERIMENTAL APPROACH: LY2033298 was investigated in vitro in CHO cells stably expressing human or mouse M(4) mAChRs, using assays of agonist-induced ERK1/2 or GSK-3alpha phosphorylation, [(35) S]-GTPgammaS binding, or effects on equilibrium binding of [(3) H]-NMS and ACh. The in vivo actions of LY2033298 were investigated in a mouse model of amphetamine-induced locomotor activity. The function of LY2033298 was examined in combination with ACh, oxotremorine or xanomeline. KEY
RESULTS: LY2033298 had similar affinities for the human and mouse M(4) mAChRs. However, LY2033298 had a lower positive co-operativity with ACh at the mouse relative to the human M(4) mAChR. At the mouse M(4) mAChR, LY2033298 showed higher co-operativity with oxotremorine than with ACh or xanomeline. The different degrees of co-operativity between LY2033298 and each agonist at the mouse relative to the human M(4) mAChR necessitated the co-administration of LY2033298 with oxotremorine in order to show in vivo efficacy of LY2033298. CONCLUSIONS AND IMPLICATIONS: These results provide evidence for species variability when comparing the allosteric interaction between LY2033298 and ACh at the M(4) mAChR, and also highlight how the interaction between LY2033298 and different orthosteric ligands is subject to 'probe dependence'. This has implications for the validation of allosteric modulator actions in vivo.
ESTHER : Suratman_2011_Br.J.Pharmacol_162_1659
PubMedSearch : Suratman_2011_Br.J.Pharmacol_162_1659
PubMedID: 21198541

Title : The role of transmembrane domain 3 in the actions of orthosteric, allosteric, and atypical agonists of the M4 muscarinic acetylcholine receptor - Leach_2011_Mol.Pharmacol_79_855
Author(s) : Leach K , Davey AE , Felder CC , Sexton PM , Christopoulos A
Ref : Molecular Pharmacology , 79 :855 , 2011
Abstract : Despite the discovery of a diverse range of novel agonists and allosteric modulators of the M(4) muscarinic acetylcholine (ACh) receptor (mAChR), little is known about how such ligands activate the receptor. We used site-directed mutagenesis of conserved residues in transmembrane 3 (TMIII), a key region involved in G protein-coupled receptor activation, to probe the binding and function of prototypical orthosteric mAChR agonists, allosteric modulators, and "atypical" agonists. We found that most mutations did not affect the binding of the allosteric modulators, with the exception of W108(3.28)A and L109(3.29)A (which may contribute directly to the interface between allosteric and orthosteric sites) and mutation D112(3.32)N (which may cause a global disruption of a hydrogen bond network). Although numerous mutations affected signaling, we did not identify amino acids that were important for the functional activity of any one class of agonist (orthosteric, allosteric, or atypical) to the exclusion of any others, suggesting that TMIII is key for the transmission of stimulus irrespective of the agonist. We also identified two key residues, Trp108(3.28) and Asp112(3.32), that are essential for the transmission of binding cooperativity between 3-amino-5-chloro-6-methoxy-4-methyl-thieno[2,3-b]pyridine- 2-carboxylic acid cyclopropylamide (LY2033298) and ACh. Finally, we found that LY2033298 was able to rescue functionally impaired signaling of ACh at the majority of mutants tested in a manner that was inversely correlated with the ACh signaling efficacy, indicating that a key part of the mechanism of the positive cooperativity mediated by LY2033298 on the endogenous agonist involves a global drive of the receptor toward an active conformation.
ESTHER : Leach_2011_Mol.Pharmacol_79_855
PubMedSearch : Leach_2011_Mol.Pharmacol_79_855
PubMedID: 21300722

Title : Structural determinants of allosteric agonism and modulation at the M4 muscarinic acetylcholine receptor: identification of ligand-specific and global activation mechanisms - Nawaratne_2010_J.Biol.Chem_285_19012
Author(s) : Nawaratne V , Leach K , Felder CC , Sexton PM , Christopoulos A
Ref : Journal of Biological Chemistry , 285 :19012 , 2010
Abstract : The recently identified small molecule, 3-amino-5-chloro-6-methoxy-4-methylthieno[2,3-b]pyridine-2-carboxylic acid cyclopropylamide (LY2033298), is the first selective allosteric modulator of the muscarinic acetylcholine receptors (mAChRs) that mediates both receptor activation and positive modulation of the endogenous agonist, acetylcholine (ACh), via the same allosteric site on the M(4) mAChR. We thus utilized this novel chemical tool, as well as ACh, the bitopic (orthosteric/allosteric) agonist, McN-A-343, and the clinically efficacious M(1)/M(4) mAChR-preferring agonist, xanomeline, in conjunction with site-directed mutagenesis of four different regions of the M(4) mAChR (extracellular loops 1, 2, and 3, and transmembrane domain 7), to identify regions that govern ligand-specific modes of binding, signaling, and allosteric modulation. In the first extracellular loop (E1), we identified Ile(93) and Lys(95) as key residues that specifically govern the signaling efficacy of LY2033298 and its binding cooperativity with ACh, whereas Phe(186) in the E2 loop was identified as a key contributor to the binding affinity of the modulator for the allosteric site, and Asp(432) in the E3 loop appears to be involved in the functional (activation) cooperativity between the modulator and the endogenous agonist. In contrast, the highly conserved transmembrane domain 7 residues, Tyr(439) and Tyr(443), were identified as contributing to a key activation switch utilized by all classes of agonists. These results provide new insights into the existence of multiple activation switches in G protein-coupled receptors (GPCRs), some of which can be selectively exploited by allosteric agonists, whereas others represent global activation mechanisms for all classes of ligand.
ESTHER : Nawaratne_2010_J.Biol.Chem_285_19012
PubMedSearch : Nawaratne_2010_J.Biol.Chem_285_19012
PubMedID: 20406819

Title : Molecular mechanisms of action and in vivo validation of an M4 muscarinic acetylcholine receptor allosteric modulator with potential antipsychotic properties - Leach_2010_Neuropsychopharmacology_35_855
Author(s) : Leach K , Loiacono RE , Felder CC , McKinzie DL , Mogg A , Shaw DB , Sexton PM , Christopoulos A
Ref : Neuropsychopharmacology , 35 :855 , 2010
Abstract : We recently identified LY2033298 as a novel allosteric potentiator of acetylcholine (ACh) at the M(4) muscarinic acetylcholine receptor (mAChR). This study characterized the molecular mode of action of this modulator in both recombinant and native systems. Radioligand-binding studies revealed that LY2033298 displayed a preference for the active state of the M(4) mAChR, manifested as a potentiation in the binding affinity of ACh (but not antagonists) and an increase in the proportion of high-affinity agonist-receptor complexes. This property accounted for the robust allosteric agonism displayed by the modulator in recombinant cells in assays of [(35)S]GTPgammaS binding, extracellular regulated kinase 1/2 phosphorylation, glycogen synthase kinase 3beta phosphorylation, and receptor internalization. We also found that the extent of modulation by LY2033298 differed depending on the signaling pathway, indicating that LY2033298 engenders functional selectivity in the actions of ACh. This property was retained in NG108-15 cells, which natively express rodent M(4) mAChRs. Functional interaction studies between LY2033298 and various orthosteric and allosteric ligands revealed that its site of action overlaps with the allosteric site used by prototypical mAChR modulators. Importantly, LY2033298 reduced [(3)H]ACh release from rat striatal slices, indicating retention of its ability to allosterically potentiate endogenous ACh in situ. Moreover, its ability to potentiate oxotremorine-mediated inhibition of condition avoidance responding in rodents was significantly attenuated in M(4) mAChR knockout mice, validating the M(4) mAChR as a key target of action of this novel allosteric ligand.
ESTHER : Leach_2010_Neuropsychopharmacology_35_855
PubMedSearch : Leach_2010_Neuropsychopharmacology_35_855
PubMedID: 19940843

Title : Selective muscarinic receptor agonist xanomeline as a novel treatment approach for schizophrenia - Shekhar_2008_Am.J.Psychiatry_165_1033
Author(s) : Shekhar A , Potter WZ , Lightfoot J , Lienemann J , Dube S , Mallinckrodt C , Bymaster FP , McKinzie DL , Felder CC
Ref : Am J Psychiatry , 165 :1033 , 2008
Abstract : OBJECTIVE: There are significant unmet needs in the treatment of schizophrenia, especially for the treatment of cognitive impairment, negative syndrome, and cognitive function. Preclinical data suggest that agonists with selective affinity for acetylcholine muscarinic receptors provide a potentially new mechanism to treat schizophrenia. The authors studied xanomeline, a relatively selective muscarinic type 1 and type 4 (M(1) and M(4)) receptor agonist, to determine if this agent is effective in the treatment of schizophrenia. METHOD: In this pilot study, the authors examined the efficacy of xanomeline on clinical outcomes in subjects with schizophrenia (N=20) utilizing a double-blind, placebo-controlled, 4-week treatment design. Outcome measures included the Positive and Negative Syndrome Scale (PANSS) for schizophrenia, the Brief Psychiatric Rating Scale (BPRS), the Clinical Global Impression (CGI) scale, and a test battery designed to measure cognitive function in patients with schizophrenia.
RESULTS: Subjects treated with xanomeline did significantly better than subjects in the placebo group on total BPRS scores and total PANSS scores. In the cognitive test battery, subjects in the xanomeline group showed improvements most robustly in measures of verbal learning and short-term memory function.
CONCLUSIONS: These results support further investigation of xanomeline as a novel approach to treating schizophrenia.
ESTHER : Shekhar_2008_Am.J.Psychiatry_165_1033
PubMedSearch : Shekhar_2008_Am.J.Psychiatry_165_1033
PubMedID: 18593778

Title : New insights into the function of M4 muscarinic acetylcholine receptors gained using a novel allosteric modulator and a DREADD (designer receptor exclusively activated by a designer drug) - Nawaratne_2008_Mol.Pharmacol_74_1119
Author(s) : Nawaratne V , Leach K , Suratman N , Loiacono RE , Felder CC , Armbruster BN , Roth BL , Sexton PM , Christopoulos A
Ref : Molecular Pharmacology , 74 :1119 , 2008
Abstract : The M4 muscarinic acetylcholine (ACh) receptor (mAChR) is a potential therapeutic target but characterized by a lack of subtype-selective ligands. We recently generated "designer receptors exclusively activated by a designer drug" (DREADDs), which contained mutations of two conserved orthosteric-site residues (Y113C/A203G in the M4 mAChR) that caused a loss of ACh activity but a gain in responsiveness to clozapine-N-oxide (CNO). The current study characterized the interactions of the wild type and the M4 DREADD with a range of agonists, antagonists, and the recently discovered M4 mAChR allosteric potentiator, 3-amino-5-chloro-6-methoxy-4-methyl-thieno[2,3-b]pyridine-2-carboxylic acid cyclopropylamide (LY2033298). LY2033298 displayed positive binding cooperativity with ACh, neutral cooperativity with the antagonist, [3H]quinuclidinyl benzilate, and agonism for activation of phosphorylated extracellular signal-regulated kinase (ERK) 1/2 at the wild-type M4 mAChR. LY2033298's cooperativity with clozapine or CNO was weakly positive with respect to binding but profoundly negative with respect to LY2033298 signaling. Although the DREADD mutations increased the binding and function of clozapine-like compounds, all other agonists lost the ability to activate the mutant; for the orthosteric agonists ACh and pilocarpine, this was due partly to a reduced affinity, whereas the affinity of LY2033298 or the atypical agonist 4-I-[3-chlorophenyl]carbamoyloxy)-2-butynyltrimethylammnonium chloride was unaltered. The interaction between LY2033298 and clozapine-like compounds reverted to neutral cooperativity on the DREADD, whereas LY2033298 caused a striking functional rescue of ACh potency and efficacy at the DREADD. These results provide conclusive evidence for the retention of a functional allosteric site on the M4 DREADD and highlight a role for residues Tyr113 and Ala203 in the transmission of cooperativity.
ESTHER : Nawaratne_2008_Mol.Pharmacol_74_1119
PubMedSearch : Nawaratne_2008_Mol.Pharmacol_74_1119
PubMedID: 18628403

Title : Allosteric modulation of the muscarinic M4 receptor as an approach to treating schizophrenia - Chan_2008_Proc.Natl.Acad.Sci.U.S.A_105_10978
Author(s) : Chan WY , McKinzie DL , Bose S , Mitchell SN , Witkin JM , Thompson RC , Christopoulos A , Lazareno S , Birdsall NJ , Bymaster FP , Felder CC
Ref : Proc Natl Acad Sci U S A , 105 :10978 , 2008
Abstract : Current antipsychotics provide symptomatic relief for patients suffering from schizophrenia and related psychoses; however, their effectiveness is variable and many patients discontinue treatment due to side effects. Although the etiology of schizophrenia is still unclear, a leading hypothesis implicates an imbalanced dopaminergic system. Muscarinic acetylcholine (ACh) receptors regulate dopamine levels in key areas of the brain involved in psychosis, with the M(4) subtype emerging as a key regulator of dopaminergic hyperactivity. Unfortunately, no selective small molecule tools exist to provide pharmacological validation of this hypothesis. Here, we describe the discovery of a small molecule modulator, LY2033298, that is highly selective for human M(4) receptors by virtue of targeting an allosteric site on this receptor. Pharmacological assays confirmed the selectivity of LY2033298 for the M(4) receptor and revealed the highest degree of positive allosteric enhancement of ACh potency thus far identified. Radioligand binding assays also show this compound to directly potentiate agonist binding while having minimal effects on antagonist binding. Mutational analysis identified a key amino acid (D(432)) in the third extracellular loop of the human M(4) receptor to be critical for selectivity and agonist potentiation by LY2033298. Importantly, LY2033298 was active in animal models predictive of clinical antipsychotic drug efficacy indicating its potential use as a first-in-class, selective, allosteric muscarinic antipsychotic agent.
ESTHER : Chan_2008_Proc.Natl.Acad.Sci.U.S.A_105_10978
PubMedSearch : Chan_2008_Proc.Natl.Acad.Sci.U.S.A_105_10978
PubMedID: 18678919

Title : Generation and analysis of muscarinic acetylcholine receptor knockout mice. -
Author(s) : Duttaroy A , Yamada M , Gomeza J , Zhang W , Miyakawa T , Makita R , Bymaster FP , Felder CC , Deng CX , Wess J
Ref : Cholinergic Mechanisms, CRC Press :63 , 2004

Title : New evidence for the involvement of muscarinic cholinergic receptors in psychoses. -
Author(s) : Bymaster FP , McKinzie DL , Felder CC
Ref : Cholinergic Mechanisms, CRC Press :331 , 2004

Title : Muscarinic mechanisms of antipsychotic atypicality - Bymaster_2003_Prog.Neuropsychopharmacol.Biol.Psychiatry_27_1125
Author(s) : Bymaster FP , Felder CC , Tzavara E , Nomikos GG , Calligaro DO , McKinzie DL
Ref : Prog Neuropsychopharmacol Biological Psychiatry , 27 :1125 , 2003
Abstract : The interactions of the atypical antipsychotic drugs (APD) clozapine, olanzapine, risperidone, quetiapine and ziprasidone with muscarinic receptors were reviewed. Only clozapine and olanzapine have marked affinity for muscarinic receptors in radioligand binding studies; however, the affinity of these compounds is considerably lower than classical muscarinic antagonists. Although functional assays in cell lines transfected with muscarinic receptors suggest that olanzapine and clozapine have weak partial agonist activity at muscarinic receptors, particularly M4 receptors, studies in vitro and in vivo indicate that the compounds function as antagonists. In animal studies and in humans, clozapine has pronounced antimuscarinic effects whereas olanzapine has weak antimuscarinic effects. However, olanzapine significantly occupies central muscarinic receptors in humans. Overall, the role of muscarinic receptors in the antipsychotic effects of clozapine and olanzapine is controversial and complex.
ESTHER : Bymaster_2003_Prog.Neuropsychopharmacol.Biol.Psychiatry_27_1125
PubMedSearch : Bymaster_2003_Prog.Neuropsychopharmacol.Biol.Psychiatry_27_1125
PubMedID: 14642972

Title : Role of specific muscarinic receptor subtypes in cholinergic parasympathomimetic responses, in vivo phosphoinositide hydrolysis, and pilocarpine-induced seizure activity - Bymaster_2003_Eur.J.Neurosci_17_1403
Author(s) : Bymaster FP , Carter PA , Yamada M , Gomeza J , Wess J , Hamilton SE , Nathanson NM , McKinzie DL , Felder CC
Ref : European Journal of Neuroscience , 17 :1403 , 2003
Abstract : Muscarinic agonist-induced parasympathomimetic effects, in vivo phosphoinositide hydrolysis and seizures were evaluated in wild-type and muscarinic M1-M5 receptor knockout mice. The muscarinic agonist oxotremorine induced marked hypothermia in all the knockout mice, but the hypothermia was reduced in M2 and to a lesser extent in M3 knockout mice. Oxotremorine-induced tremor was abolished only in the M2 knockout mice. Muscarinic agonist-induced salivation was reduced to the greatest extent in M3 knockout mice, to a lesser degree in M1 and M4 knockout mice, and was not altered in M2 and M5 knockout mice. Pupil diameter under basal conditions was increased only in the M3 knockout mice. Pilocarpine-induced increases in in vivo phosphoinositide hydrolysis were completely absent in hippocampus and cortex of M1 knockout mice, but in vivo phosphoinositide hydrolysis was unaltered in the M2-M5 knockout mice. A high dose of pilocarpine (300 mg/kg) caused seizures and lethality in wild-type and M2-M5 knockout mice, but produced neither effect in the M1 knockout mice. These data demonstrate a major role for M2 and M3 muscarinic receptor subtypes in mediating parasympathomimetic effects. Muscarinic M1 receptors activate phosphoinositide hydrolysis in cortex and hippocampus of mice, consistent with the role of M1 receptors in cognition. Muscarinic M1 receptors appear to be the only muscarinic receptor subtype mediating seizures.
ESTHER : Bymaster_2003_Eur.J.Neurosci_17_1403
PubMedSearch : Bymaster_2003_Eur.J.Neurosci_17_1403
PubMedID: 12713643

Title : Muscarinic receptor subtypes mediating central and peripheral antinociception studied with muscarinic receptor knockout mice: a review - Wess_2003_Life.Sci_72_2047
Author(s) : Wess J , Duttaroy A , Gomeza J , Zhang W , Yamada M , Felder CC , Bernardini N , Reeh PW
Ref : Life Sciences , 72 :2047 , 2003
Abstract : To gain new insight into the physiological and pathophysiological roles of the muscarinic cholinergic system, we generated mutant mouse strains deficient in each of the five muscarinic acetylcholine receptor subtypes (M(1)-M(5)). In this chapter, we review a set of recent studies dealing with the identification of the muscarinic receptor subtypes mediating muscarinic agonist-dependent analgesic effects by central and peripheral mechanisms. Most of these studies were carried out with mutant mouse strains lacking M(2) or/and M(4) muscarinic receptors. It is well known that administration of centrally active muscarinic agonists induces pronounced analgesic effects. To identify the muscarinic receptors mediating this activity, wild-type and muscarinic receptor mutant mice were injected with the non-subtype-selective muscarinic agonist, oxotremorine (s.c., i.t., and i.c.v.), and analgesic effects were assessed in the tail-flick and hot-plate tests. These studies showed that M(2) receptors play a key role in mediating the analgesic effects of oxotremorine, both at the spinal and supraspinal level. However, studies with M(2)/M(4) receptor double KO mice indicated that M(4) receptors also contribute to this activity. Recent evidence suggests that activation of muscarinic receptors located in the skin can reduce the sensitivity of peripheral nociceptors. Electrophysiological and neurochemical studies with skin preparations from muscarinic receptor mutant mice indicated that muscarine-induced peripheral antinociception is mediated by M(2) receptors. Since acetylcholine is synthesized and released by different cell types of the skin, it is possible that non-neuronally released acetylcholine plays a role in modulating peripheral nociception. Our results highlight the usefulness of muscarinic receptor mutant mice to shed light on the functional roles of acetylcholine released from both neuronal and non-neuronal cells.
ESTHER : Wess_2003_Life.Sci_72_2047
PubMedSearch : Wess_2003_Life.Sci_72_2047
PubMedID: 12628455

Title : M1 muscarinic receptor signaling in mouse hippocampus and cortex - Porter_2002_Brain.Res_944_82
Author(s) : Porter AC , Bymaster FP , DeLapp NW , Yamada M , Wess J , Hamilton SE , Nathanson NM , Felder CC
Ref : Brain Research , 944 :82 , 2002
Abstract : The five subtypes (M1-M5) of muscarinic acetylcholine receptors signal through G(alpha)(q) or G(alpha)(i)/G(alpha)(o). M1, M3 and M5 receptors couple through G(alpha)(q) and function predominantly as postsynaptic receptors in the central nervous system. M1 and M3 receptors are localized to brain regions involved in cognition, such as hippocampus and cortex, but their relative contribution to function has been difficult to ascertain due to the lack of subtype specific ligands. A functional and genetic approach was used to identify the predominant muscarinic receptor subtype(s) mediating responses in mouse hippocampus and cortex, as well as the relative degree of spare muscarinic receptors in hippocampus. The nonselective muscarinic agonist oxotremorine-M stimulated G(alpha)(q)/11-specific GTP-gamma-35S binding in a concentration dependent manner with a Hill slope near unity in wild type mouse hippocampus and cortex. Muscarinic receptor stimulated G(alpha)(q)/11-specific GTP-gamma-35S binding was virtually abolished in both the hippocampus and cortex of M1 receptor knockout (KO) mice. In contrast, there was no loss of signaling in M3 receptor KO mice in either brain region. Muscarinic receptor reserve in wildtype mouse hippocampus was measured by Furchgott analysis after partial receptor alkylation with propylbenzylcholine mustard. Occupation of just 15% of the M1 receptors in mouse hippocampus was required for maximal efficacy of oxotremorine-M-stimulated GTP-gamma-35S binding indicating a substantial level of spare receptors. These findings support a role for the M1 receptor subtype as the primary G(alpha)(q)/11-coupled muscarinic receptor in mouse hippocampus and cortex.
ESTHER : Porter_2002_Brain.Res_944_82
PubMedSearch : Porter_2002_Brain.Res_944_82
PubMedID: 12106668

Title : Evaluation of muscarinic agonist-induced analgesia in muscarinic acetylcholine receptor knockout mice - Duttaroy_2002_Mol.Pharmacol_62_1084
Author(s) : Duttaroy A , Gomeza J , Gan JW , Siddiqui N , Basile AS , Harman WD , Smith PL , Felder CC , Levey AI , Wess J
Ref : Molecular Pharmacology , 62 :1084 , 2002
Abstract : Centrally active muscarinic agonists display pronounced analgesic effects. Identification of the specific muscarinic acetylcholine receptor (mAChR) subtype(s) mediating this activity is of considerable therapeutic interest. To examine the roles of the M(2) and M(4) receptor subtypes, the two G(i)/G(o)-coupled mAChRs, in mediating agonist-dependent antinociception, we generated a mutant mouse line deficient in both M(2) and M(4) mAChRs [M(2)/M(4) double-knockout (KO) mice]. In wild-type mice, systemic, intrathecal, or intracerebroventricular administration of centrally active muscarinic agonists resulted in robust analgesic effects, indicating that muscarinic analgesia can be mediated by both spinal and supraspinal mechanisms. Strikingly, muscarinic agonist-induced antinociception was totally abolished in M(2)/M(4) double-KO mice, independent of the route of application. The nonselective muscarinic agonist oxotremorine showed reduced analgesic potency in M(2) receptor single-KO mice, but retained full analgesic activity in M(4) receptor single-KO mice. In contrast, two novel muscarinic agonists chemically derived from epibatidine, CMI-936 and CMI-1145, displayed reduced analgesic activity in both M(2) and M(4) receptor single-KO mice, independent of the route of application. Radioligand binding studies indicated that the two CMI compounds, in contrast to oxotremorine, showed >6-fold higher affinity for M(4) than for M(2) receptors, providing a molecular basis for the observed differences in agonist activity profiles. These data provide unambiguous evidence that muscarinic analgesia is exclusively mediated by a combination of M(2) and M(4) mAChRs at both spinal and supraspinal sites. These findings should be of considerable relevance for the development of receptor subtype-selective muscarinic agonists as novel analgesic drugs.
ESTHER : Duttaroy_2002_Mol.Pharmacol_62_1084
PubMedSearch : Duttaroy_2002_Mol.Pharmacol_62_1084
PubMedID: 12391271

Title : Role of the cholinergic muscarinic system in bipolar disorder and related mechanism of action of antipsychotic agents - Bymaster_2002_Mol.Psychiatry_7 Suppl 1_S57
Author(s) : Bymaster FP , Felder CC
Ref : Mol Psychiatry , 7 Suppl 1 :S57 , 2002
Abstract : The evidence for the involvement of cholinergic muscarinic receptors in mania and depression is reviewed. Small pilot trials with cholinesterase inhibitors and muscarinic agonists suggest that stimulation of muscarinic receptors may produce an antimanic effect, possibly by activation of muscarinic M(4) receptors. It is concluded that it is not likely that currently used mood stabilizers, such as lithium, valproic acid and carbamazepine, work directly through muscarinic receptor mechanisms. Furthermore, the evidence indicates that antipsychotic agents used for mania are working through the common mechanism of antagonism of dopamine D(2) receptors, and interactions with muscarinic receptors do not play a key role. Finally, it is hypothesized that olanzapine has robust antimanic activity, due to blockade of dopamine D(2) receptors and antagonism of other monoaminergic receptors. Olanzapine may normalize mood due to antidepressant-like activities, such as 5-HT(2A) receptor antagonism and increasing cortical norepinephrine and dopamine.
ESTHER : Bymaster_2002_Mol.Psychiatry_7 Suppl 1_S57
PubMedSearch : Bymaster_2002_Mol.Psychiatry_7 Suppl 1_S57
PubMedID: 11986996

Title : Poster: The M1 muscarinic receptor subtype mediates Gq coupling in mouse hippocampus and pre-frontal cortex -
Author(s) : Porter AC , Bymaster FP , Nathanson NM , Hamilton SE , Carter PA , DeLapp NW , Felder CC
Ref : Life Sciences , 68 :2624 , 2001

Title : Receptor reserve of phosphoinositide-coupled muscarinic receptors in mouse hippocampus in vivo - Bymaster_2001_Brain.Res_916_165
Author(s) : Bymaster FP , Carter PA , DeLapp NW , Calligaro DO , Felder CC
Ref : Brain Research , 916 :165 , 2001
Abstract : The ability of the partial muscarinic agonist pilocarpine to increase in vivo phosphoinositide (PI) hydrolysis in mouse brain was compared to two full agonists. Pilocarpine increased in vivo phosphoinositide (PI) hydrolysis in cortex, striatum, and to the greatest extent in the hippocampus. Pilocarpine injected either subcutaneously or intracerebroventricularly robustly increased in vivo PI hydrolysis in hippocampus up to 500% of control levels and the increases were blocked by the muscarinic antagonist scopolamine. The increases in vivo PI hydrolysis induced by pilocarpine were 60-75% of the magnitude of the full muscarinic agonists oxotremorine-M and cis-dioxolane. The muscarinic M(1) preferring antagonist pirenzepine potently blocked pilocarpine-induced increases in in vivo PI hydrolysis, consistent with the increase being mediated by M(1) receptors. Since pilocarpine is a relatively weak partial agonist, these data suggest a substantial level of receptor reserve for the PI response in mouse hippocampus.
ESTHER : Bymaster_2001_Brain.Res_916_165
PubMedSearch : Bymaster_2001_Brain.Res_916_165
PubMedID: 11597604

Title : Poster: Evaluation of muscarinic agonist-induced analgesia in muscarinic receptor knockout mice -
Author(s) : Duttaroy A , Gomeza J , Gan JW , Basile AS , Harman WD , Smith PL , Felder CC , Levey AI , Wess J
Ref : Life Sciences , 68 :2631 , 2001

Title : Poster: Enhanced sensitivity to the locomotor-activating and disrupting effects of phencyclidine in M4 muscarinic receptor knock-out mice -
Author(s) : Felder CC , Bymaster FP , Wess J , Zhang L , Shaw DB , McKinzie DL
Ref : Life Sciences , 68 :2636 , 2001

Title : Neostriatal muscarinic receptor subtypes involved in the generation of tremulous jaw movements in rodents implications for cholinergic involvement in parkinsonism - Salamone_2001_Life.Sci_68(22-23)_2579
Author(s) : Salamone JD , Correa M , Carlson BB , Wisniecki A , Mayorga AJ , Nisenbaum E , Nisenbaum L , Felder CC
Ref : Life Sciences , 68 :2579 , 2001
Abstract : Several studies have shown that a number of pharmacological and neurochemical conditions in rats can induce jaw movements that are described as "vacuous" or "tremulous". For several years, there has been some debate about the clinical significance of various drug-induced oral motor syndromes. Nevertheless, considerable evidence now indicates that the non-directed, chewing-like movements induced by cholinomimetics have many of the characteristics of parkinsonian tremor. These movements are characterized largely by vertical deflections of the jaw, which occur in the same 3-7 Hz peak frequency that is typical of parkinsonian tremor. Cholinomimetic-induced tremulous jaw movements are suppressed by a number of different antiparkinsonian drugs, including scopolamine, benztropine, L-DOPA, apomorphine, bromocriptine, ropinirole, pergolide, amantadine, diphenhydramine and clozapine. A combination of anatomical and pharmacological research in rats has implicated M4 receptors in the ventrolateral neostriatum in the generation of tremulous jaw movements. Mice also show cholinomimetic-induced jaw movements, and M4 receptor knockout mice demonstrate subtantially reduced levels of jaw movement activity, as well as increased locomotion. Taken together, these data are consistent with the hypothesis that a centrally-acting M4 antagonist may be useful as a treatment for parkinsonian symptoms, including tremor.
ESTHER : Salamone_2001_Life.Sci_68(22-23)_2579
PubMedSearch : Salamone_2001_Life.Sci_68(22-23)_2579
PubMedID: 11392629

Title : Investigations into the physiological role of muscarinic M2 and M4 muscarinic and M4 receptor subtypes using receptor knockout mice - Bymaster_2001_Life.Sci_68(22-23)_2473
Author(s) : Bymaster FP , Carter PA , Zhang L , Falcone JF , Stengel PW , Cohen ML , Shannon HE , Gomeza J , Wess J , Felder CC
Ref : Life Sciences , 68 :2473 , 2001
Abstract : Determination of muscarinic agonist-induced parasympathomimetic effects in wild type and M2 and M4 muscarinic receptor knockout mice revealed that M2 receptors mediated tremor and hypothermia, but not salivation. The M4 receptors seem to play a modest role in salivation, but did not alter hypothermia and tremor. In the M2 knockout mice, agonist-induced bradycardia in isolated spontaneously beating atria was completely absent compared to their wild type litter mates, whereas agonist-induced bradycardia was similar in the M4 knockout and wild type mice. The potency of carbachol to stimulate contraction of isolated stomach fundus, urinary bladder and trachea was reduced by a factor of about 2 in the M2 knockout mice, but was unaltered in the M4 knockout mice. The binding of the muscarinic agonist, [3H]-oxotremorine-M, was reduced in cortical tissue from the M2 knockout mice and to a lesser extent from the M4 knockout mice, and was reduced over 90% in the brain stem of M2 knockout mice. The data demonstrate the usefulness of knockout mice in determining the physiological function of peripheral and central muscarinic receptors.
ESTHER : Bymaster_2001_Life.Sci_68(22-23)_2473
PubMedSearch : Bymaster_2001_Life.Sci_68(22-23)_2473
PubMedID: 11392615

Title : The muscarinic agonist xanomeline increases monoamine release and immediate early gene expression in the rat prefrontal cortex - Perry_2001_Biol.Psychiatry_49_716
Author(s) : Perry KW , Nisenbaum LK , George CA , Shannon HE , Felder CC , Bymaster FP
Ref : Biological Psychiatry , 49 :716 , 2001
Abstract : BACKGROUND: The muscarinic agonist xanomeline has been shown to reduce antipsychotic-like behaviors in patients with Alzheimer's disease. Because atypical antipsychotic agents increase dopamine release in prefrontal cortex and induce immediate early gene expression in prefrontal cortex and nucleus accumbens, the effect of xanomeline was determined on these indices.
METHODS: The effect of xanomeline on extracellular levels of monoamines in brain regions was determined using a microdialysis technique, and changes in expression of the immediate early genes c-fos and zif/268 in brain regions were evaluated using in situ hybridization histochemistry.
RESULTS: Xanomeline increased extracellular levels of dopamine in prefrontal cortex and nucleus accumbens but not in striatum. Xanomeline increased expression of c-fos and zif/268 in prefrontal cortex and nucleus accumbens. There was no change in immediate early gene expression in striatum.
CONCLUSIONS: Xanomeline increased extracellular levels of dopamine, which is similar to the effects of the atypical antipsychotics clozapine and olanzapine. The regional pattern of immediate early gene expression induced by xanomeline resembled that of atypical antipsychotic agents. Based on the antipsychotic-like activity of xanomeline in Alzheimer's patients and the similarity to atypical antipsychotic agents, we suggest that xanomeline may be a novel antipsychotic agent.
ESTHER : Perry_2001_Biol.Psychiatry_49_716
PubMedSearch : Perry_2001_Biol.Psychiatry_49_716
PubMedID: 11313039

Title : Elucidating the role of muscarinic receptors in psychosis - Felder_2001_Life.Sci_68(22-23)_2605
Author(s) : Felder CC , Porter AC , Skillman TL , Zhang L , Bymaster FP , Nathanson NM , Hamilton SE , Gomeza J , Wess J , McKinzie DL
Ref : Life Sciences , 68 :2605 , 2001
Abstract : Muscarinic receptors have been implicated in the regulation of cognition and psychosis based on pharmacological evidence from pre-clinical and clinical studies. Muscarinic agonists have shown promise in the clinic in improving cognition and reducing psychotic episodes in Alzheimer's patients. However, lack of selective muscarinic ligands has limited their use due to troublesome side effects observed at higher doses. Without selective ligands, it has been difficult to assign a specific muscarinic receptor subtype to these high order mental processes. Recent development of muscarinic receptor knockout mice has provided additional tools to investigate cognition and psychosis in behavioral assays and to determine the receptor subtypes associated with parasympathomimetic physiology. Biochemical studies indicate that the M1 receptor plays a significant role in regulating G alpha q-mediated signal transduction in the hippocampus and cortex. Behavioral studies suggest that the M4 receptor is involved in movement regulation and prepulse inhibition of the startle reflex, a measure of attention. These findings support a role for the development of M1 and M4 receptor agonists for diseases in which symptoms include cognitive impairment and psychotic behaviors.
ESTHER : Felder_2001_Life.Sci_68(22-23)_2605
PubMedSearch : Felder_2001_Life.Sci_68(22-23)_2605
PubMedID: 11392633

Title : Generation and pharmacological analysis of M2 and M4 muscarinic receptor knockout mice - Gomeza_2001_Life.Sci_68(22-23)_2457
Author(s) : Gomeza J , Zhang L , Kostenis E , Felder CC , Bymaster FP , Brodkin J , Shannon HE , Xia B , Duttaroy A , Deng CX , Wess J
Ref : Life Sciences , 68 :2457 , 2001
Abstract : Muscarinic acetylcholine receptors (M1-M5) play important roles in the modulation of many key functions of the central and peripheral nervous system. To explore the physiological roles of the two Gi-coupled muscarinic receptors, we disrupted the M2 and M4 receptor genes in mice by using a gene targeting strategy. Pharmacological and behavioral analysis of the resulting mutant mice showed that the M2 receptor subtype is critically involved in mediating three of the most striking central muscarinic effects, tremor, hypothermia, and analgesia. These studies also indicated that M4 receptors are not critically involved in these central muscarinic responses. However, M4 receptor-deficient mice showed an increase in basal locomotor activity and greatly enhanced locomotor responses following drug-induced activation of D1 dopamine receptors. This observation is consistent with the concept that M4 receptors exert inhibitory control over D1 receptor-mediated locomotor stimulation, probably at the level of striatal projection neurons where the two receptors are known to be coexpressed. These findings emphasize the usefulness of gene targeting approaches to shed light on the physiological and pathophysiological roles of the individual muscarinic receptor subtypes.
ESTHER : Gomeza_2001_Life.Sci_68(22-23)_2457
PubMedSearch : Gomeza_2001_Life.Sci_68(22-23)_2457
PubMedID: 11392613

Title : A current review of olanzapine's safety in the geriatric patient: from pre-clinical pharmacology to clinical data - Kennedy_2001_Int.J.Geriatr.Psychiatry_16 Suppl 1_S33
Author(s) : Kennedy JS , Bymaster FP , Schuh L , Calligaro DO , Nomikos G , Felder CC , Bernauer M , Kinon BJ , Baker RW , Hay D , Roth HJ , Dossenbach M , Kaiser C , Beasley CM , Holcombe JH , Effron MB , Breier A
Ref : Int J Geriatr Psychiatry , 16 Suppl 1 :S33 , 2001
Abstract : OBJECTIVE: Olanzapine (OLZ) is unique among currently available antipsychotic medications in its antagonism of a range of receptor systems including dopamine, norepinephrine, serotonin, acetylcholine, and histamine. Olanzapine's mechanistic complexity provides a broad efficacy profile in patients with schizophrenia and acute, pure or mixed mania. Patients experience symptomatic relief of mania, anxiety, hallucinations, delusions, and agitation/aggression and reduced depressive, negative, and some cognitive symptoms. This paper will review the safety profile of OLZ, focusing on the elderly, where data are available. METHOD: Preclinical and clinical studies of OLZ are reviewed, with emphasis on its possible effects on the cholinergic system and the histamine H(1) receptor. Weight change and related metabolic considerations, cardiac and cardiovascular safety, and motor function during treatment with OLZ are also reviewed. RESULTS AND CONCLUSION: In vitro receptor characterization methods, when done using physiologically relevant conditions allow accurate prediction of the relatively low rate of anticholinergic-like adverse events, extrapyramidal symptoms, and cardiovascular adverse events during treatment with OLZ. Currently available clinical data suggest olanzapine is predictably safe in treating adult patients of any age with schizophrenia and acute bipolar mania, as well as in treatment of patients with some types of neurodegenerative disorders.
ESTHER : Kennedy_2001_Int.J.Geriatr.Psychiatry_16 Suppl 1_S33
PubMedSearch : Kennedy_2001_Int.J.Geriatr.Psychiatry_16 Suppl 1_S33
PubMedID: 11748788

Title : Therapeutic opportunities for muscarinic receptors in the central nervous system -
Author(s) : Felder CC , Bymaster FP , Ward JS , DeLapp NC
Ref : Journal of Medicinal Chemistry , 43 :4333 , 2000
PubMedID: 11087557

Title : Poster: Comparison of GTP-gamma-35S and 3H-NMS binding in CHO cells stably expressing human M1-M5 muscarinic receptors using scintillation proximity assays -
Author(s) : Sawyer BD , Silbernagel A , McKinzie JB , Falcone J , DeLapp NW , Felder CC
Ref : Life Sciences , 64 :568 , 1999

Title : Poster: A study of M1 receptor coupling to inhibitory G proteins in CHO cells via GTPS binding determined with anti-G protein antibodies -
Author(s) : DeLapp NW , McClure D , McKinzie J , Silbernagel A , Sawyer BD , Felder CC
Ref : Life Sciences , 64 :569 , 1999

Title : Poster: Muscarinic receptor-activated arachidonic acid release is enhanced by cytokines in A2058 human melanoma cells -
Author(s) : Wood MW , Briley EM , Felder CC
Ref : Life Sciences , 64 :566 , 1999

Title : Poster: An anti-G protein scintillation proximity assay for M1-M5 receptor-mediated GTPS binding -
Author(s) : McKinzie JH , Sawyer BD , Felder CC , Falcone J , Silbernagel A , McClure D , DeLapp NW
Ref : Life Sciences , 64 :570 , 1999

Title : 1-(1,2,5-Thiadiazol-4-yl)-4-azatricyclo[,6)]heptanes as new potent muscarinic M1 agonists: structure-activity relationship for 3-aryl-2-propyn-1-yloxy and 3-aryl-2-propyn-1-ylthio derivatives - Jeppesen_1999_J.Med.Chem_42_1999
Author(s) : Jeppesen L , Olesen PH , Hansen L , Sheardown MJ , Thomsen C , Rasmussen T , Jensen AF , Christensen MS , Rimvall K , Ward JS , Whitesitt CA , Calligaro DO , Bymaster FP , DeLapp NW , Felder CC , Shannon HE , Sauerberg P
Ref : Journal of Medicinal Chemistry , 42 :1999 , 1999
Abstract : Two new series of 1-(1,2,5-thiadiazol-4-yl)-4-azatricyclo[, 6)]heptanes were synthesized and evaluated for their in vitro activity in cell lines transfected with either the human M1 or M2 receptor. 3-Phenyl-2-propyn-1-yloxy and -1-ylthio analogues substituted with halogen in the meta position showed high functional potency, efficacy, and selectivity toward the M1 receptor subtype. A quite unique functional M1 receptor selectivity was observed for compounds 8b, 8d, 8f, 9b, 9d, and 9f. Bioavailability studies in rats indicated an oral bioavailability of about 20-30%, with the N-oxide as the only detected metabolite.
ESTHER : Jeppesen_1999_J.Med.Chem_42_1999
PubMedSearch : Jeppesen_1999_J.Med.Chem_42_1999
PubMedID: 10354408

Title : Poster: New potent muscarinic 1,2,5-thiadiazole-azacycles and dimers hereof -
Author(s) : Jeppesen L , Sauerberg P , Olesen PH , Sheardown MJ , Thomsen C , Rasmussen T , Fink-Jensen A , Christensen MS , Ward JS , Whitesitt C , Calligaro DO , Bymaster FP , DeLapp NW , Felder CC , Shannon HE
Ref : Life Sciences , 64 :555 , 1999

Title : Muscarinic receptor-activated arachidonic acid release is enhanced by cytokines in A2058 human melanoma cells -
Author(s) : Felder CC , Joon JH , Briley EM , Wood MW
Ref : Journal de Physiologie (Paris) , 92 :431 , 1998

Title : Cytosolic phospholipase A2 (cPLA2) distribution in murine brain and functional studies indicate that cPLA2 does not participate in muscarinic receptor-mediated signaling in neurons -
Author(s) : Lamar T , Lautens LL , Chiou XG , Sharp JD , Young III WS , Sprague DL , Ross LS , Felder CC
Ref : Journal de Physiologie (Paris) , 92 :454 , 1998

Title : Muscarinic receptor activated Ca2+ channels in non-excitable cells - Singer-Lahat_1996_Prog.Brain.Res_109_195
Author(s) : Singer-Lahat D , Rojas E , Felder CC
Ref : Prog Brain Res , 109 :195 , 1996
Abstract : We have provided preliminary characterization of a single channel Ca2+ conductance in CHO cells. We have demonstrated that the channel conducts Ca2+, is regulated by m5 receptors, is voltage-independent, has an extremely low conductance, and is second messenger-independent. This channel may be the receptor-operated channel required for downstream activation of several signaling events. It is not known what other cell types express the channel or if it is one of a larger group of related channels. It seems likely that Ca2+ influx-dependent signaling pathways, activated by the muscarinic m5 receptor, would utilize a plasma membrane resident Ca2+ channel to provide a steady source of Ca2+ from outside the cell. The transient nature of IP3-activated increases in intracellular Ca2+ make it an unlikely source of the sustained Ca2+ rise required for phospholipase regulation. This is especially surprising, since levels of intracellular Ca2+ achieved from the release of intracellular Ca2+ stores can be at least one order of magnitude higher than those achieved from extracellular influx (Berridge, 1993). The phospholipase A2 and phospholipase D involved in muscarinic receptor-mediated signaling have not been purified or cloned. It is possible that receptor-activated and Ca2+ influx-dependent phospholipases are integral membrane proteins located adjacent to both receptors and channels. The phospholipases may also translocate to the membrane following activation where they would gain access to the continuous Ca2+ flow. Purification and cloning of this and other related channels should provide better insight into their role in cell signaling.
ESTHER : Singer-Lahat_1996_Prog.Brain.Res_109_195
PubMedSearch : Singer-Lahat_1996_Prog.Brain.Res_109_195
PubMedID: 9009707

Title : Independent induction of morphological transformation of CHO cells by receptor-activated cyclic AMP synthesis or by receptor-operated calcium influx - Singer-Lahat_1996_Biochem.Pharmacol_51_495
Author(s) : Singer-Lahat D , Ma AL , Felder CC
Ref : Biochemical Pharmacology , 51 :495 , 1996
Abstract : Morphological transformation of Chinese hamster ovary (CHO) cells can be induced by exogenous addition of cyclic AMP (cAMP) or through the stimulation of G protein-coupled receptors ectopically expressed in these cells. The morphological transformation has been shown to represent a phenotypic suppression of CHO cell tumorigenic potential. Studies were undertaken to determine which receptor-activated signal transduction pathway initiates the progression from a tumorigenic to a non-tumorigenic phenotype. Stimulation of CHO cells expressing the dopamine D1 receptor (CHOD1) with a D1 selective agonist, SKF38393, resulted in an increase in cAMP accumulation which correlated with morphologic transformation. SKF38393 had no effect on intracellular calcium levels, arguing against a requirement for phospholipase C or calcium mobilization in the D1-stimulated morphology change. In contrast, stimulation of muscarinic m5 (CHOm5) or vasopressin V1a (CHOV1a) receptors expressed in CHO cells with carbachol or arginine vasopressin (AVP), respectively, did not result in an increase in intracellular calcium and a morphology change. The time course of carbachol-stimulated calcium influx correlated with the time course of morphological transformation, but not with carbachol-stimulated cAMP or inositol, 1,4,5-trisphosphate (IP3) accumulation. Furthermore, no increase in cAMP accumulation was observed in AVP-stimulated CHOV1a cells, suggesting a cAMP-independent stimulation of the transformation process. Carbachol-stimulated CHO cells expressing the m2 muscarinic receptor (CHOm2) failed to undergo a morphological transformation, yet released IP3. Therefore, phospholipase C-mediated signal transduction is not sufficient for the morphological transformation of CHO cells. It appears that receptor-stimulated morphologic transformation of CHO cells can be induced via two independent signaling pathways, mediated by adenylate cyclase or receptor-operated calcium channels.
ESTHER : Singer-Lahat_1996_Biochem.Pharmacol_51_495
PubMedSearch : Singer-Lahat_1996_Biochem.Pharmacol_51_495
PubMedID: 8619896

Title : Muscarinic acetylcholine receptor subtypes associated with release of Alzheimer amyloid precursor derivatives activate multiple signal transduction pathways - Felder_1993_Ann.N.Y.Acad.Sci_695_15
Author(s) : Felder CC , Ma AL , Briley EM , Axelrod J
Ref : Annals of the New York Academy of Sciences , 695 :15 , 1993
Abstract : Five subtypes of muscarinic acetylcholine receptors have been identified and designated m1-m5. The m1 and m3 receptors have recently been shown to stimulate APP processing. The m1 and m3 receptors couple to a variety of signal transduction pathways in both tissue slices and a variety of cell lines endogenously expressing either or both subtypes. In contrast, the m2 and m4 receptors have been primarily associated with inhibition of adenylate cyclase. We have transfected all five subtypes of muscarinic receptors into a variety of mammalian cell lines in order to investigate the signaling associated with single receptor subtypes. The m1, m3, or m5 receptors stimulate phospholipase A2, C, and D, adenylate cyclase, receptor-operated calcium channels, and tyrosine kinase activity simultaneously. The m2 or m4 receptor inhibits cAMP accumulation and augments a previously stimulated release of arachidonic acid and calcium influx.
ESTHER : Felder_1993_Ann.N.Y.Acad.Sci_695_15
PubMedSearch : Felder_1993_Ann.N.Y.Acad.Sci_695_15
PubMedID: 8239275

Title : Tumor-suppressor function of muscarinic acetylcholine receptors is associated with activation of receptor-operated calcium influx - Felder_1993_Proc.Natl.Acad.Sci.U.S.A_90_1706
Author(s) : Felder CC , MacArthur L , Ma AL , Gusovsky F , Kohn EC
Ref : Proc Natl Acad Sci U S A , 90 :1706 , 1993
Abstract : Several members of the family of guanine nucleotide-binding protein (G protein)-coupled receptors have recently been shown to induce agonist-dependent foci development in NIH 3T3 cells and tumors in nude mice. We selected the five subtypes of the muscarinic acetylcholine receptor family to investigate their role in tumor suppression. When transfected and expressed in CHO-K1 Chinese hamster ovary cells, m1, m3, and m5 muscarinic acetylcholine receptor activation resulted in a morphology change. Receptor activation did not slow or inhibit monolayer growth of CHOm5 cells in culture but markedly inhibited density-independent growth in soft agar and suppressed tumor formation in nude mice. Receptor-mediated tumor suppression was found to be agonist-dependent and reversible and was blocked with a muscarinic receptor antagonist. Of the five signaling pathways associated with the m1, m3, and m5 receptors, only receptor-operated, and inositol trisphosphate-independent, calcium influx was found to correlate with inhibition of tumorigenicity. These data suggest a pivotal role for inositol trisphosphate-independent receptor-regulated calcium homeostasis in CHO-K1 tumor suppression.
ESTHER : Felder_1993_Proc.Natl.Acad.Sci.U.S.A_90_1706
PubMedSearch : Felder_1993_Proc.Natl.Acad.Sci.U.S.A_90_1706
PubMedID: 7680475

Title : Receptor-coupled amyloid precursor protein processing - Nitsch_1993_Ann.N.Y.Acad.Sci_695_122
Author(s) : Nitsch RM , Slack BE , Farber SA , Borghesani PR , Schulz JG , Kim C , Felder CC , Growdon JH , Wurtman RJ
Ref : Annals of the New York Academy of Sciences , 695 :122 , 1993
Abstract : The family of beta-amyloid protein precursors (APP) can be processed via several alternative proteolytic pathways. Some generate potentially amyloidogenic APP derivatives, whereas others preclude the formation of such fragments. The cellular mechanisms regulating the relative activities of these pathways are thus important in determining the factors contributing to the formation of amyloidogenic APP derivatives. In order to investigate whether cell-surface receptor activity can regulate APP processing, HEK 293 cell lines stably expressing human muscarinic acetylcholine receptors (mAChR; subtypes m1, m2, m3, m4) were stimulated with the muscarinic agonist carbachol, and the release of APP derivatives was measured. Carbachol increased the release of large amino-terminal APP-fragments 4- to 6-fold in cell lines expressing the m1 or m3 receptors but not in those expressing m2 or m4 subtypes. This increase was blocked by various protein kinase inhibitors and mimicked by phorbol esters, indicating that it is mediated by protein kinase activation, presumably by protein kinase C (PKC). To determine whether additional cell-surface receptor types linked to this signal transduction pathway could also regulate APP processing, we stimulated differentiated PC-12 cells with bradykinin and found that this neuropeptide also increased the secretion of amino-terminal APP derivatives. We next investigated the possibility that neuronal depolarization might affect APP processing in mammalian brain. Electrically stimulated rat hippocampal slices released two times more amino-terminal APP derivatives than unstimulated control slices. This release increased with increasing stimulation frequencies in the physiological firing range of hippocampal pyramidal cells, and was blocked by tetrodotoxin. These results suggest that, in brain, APP processing is regulated by neuronal activity.
ESTHER : Nitsch_1993_Ann.N.Y.Acad.Sci_695_122
PubMedSearch : Nitsch_1993_Ann.N.Y.Acad.Sci_695_122
PubMedID: 8239269

Title : The antiproliferative and antimetastatic compound L651582 inhibits muscarinic acetylcholine receptor-stimulated calcium influx and arachidonic acid release - Felder_1991_J.Pharmacol.Exp.Ther_257_967
Author(s) : Felder CC , Ma AL , Liotta LA , Kohn EC
Ref : Journal of Pharmacology & Experimental Therapeutics , 257 :967 , 1991
Abstract : L651582, a carboxyamide-amino-imidazole, was shown previously to have antiproliferative and antimetastatic properties at low micromolar concentrations; yet little is known about its cellular mechanism(s) of action. L651582 was tested for its ability to block receptor-stimulated calcium influx, arachidonic acid release, inositol phosphate and cyclic AMP (cAMP) generation. These signal transduction pathways are activated by muscarinic receptors transfected and expressed in Chinese hamster ovary cells. L651582 blocked muscarinic m5 receptor-stimulated 45Ca++ influx and release of arachidonic acid at low micromolar concentrations. Muscarinic receptor-stimulated release of arachidonic acid was shown previously to be dependent on calcium influx and not intracellular calcium release suggesting L651582 may be useful as calcium channel blocker. At low micromolar concentrations, L651582 had little effect on muscarinic m5 receptor-stimulated release of inositol phosphates or cAMP accumulation. Moreover, L651582 had little effect on muscarinic m2 receptor-mediated inhibition of forskolin-stimulated cAMP accumulation. Above 10 microM, L651582 inhibited all second messenger pathways tested and inhibited cell growth, suggesting its action may be less specific and toxic at these concentrations.
ESTHER : Felder_1991_J.Pharmacol.Exp.Ther_257_967
PubMedSearch : Felder_1991_J.Pharmacol.Exp.Ther_257_967
PubMedID: 1646332

Title : Muscarinic receptors mediate the release of arachidonic acid from spinal cord and hippocampal neurons in primary culture - Kanterman_1990_Neurosci.Lett_118_235
Author(s) : Kanterman RY , Ma AL , Briley EM , Axelrod J , Felder CC
Ref : Neuroscience Letters , 118 :235 , 1990
Abstract : Muscarinic receptors are involved in CNS neurotransmissions and have been shown to transduce their message by modulating cAMP, calcium, inositol phosphates, and more recently, by liberating arachidonic acid via phospholipase A1. We have previously shown that the alpha 1-adrenergic and 5-HT2 serotonergic neurotransmitter receptors cause the release of arachidonic acid from spinal cord and hippocampal neurons, respectively, in primary culture. In this study, we demonstrated a muscarinic receptor-mediated release of arachidonic acid in these two neural segments which occurred independent of phosphatidylinositol-specific phospholipase C. This release of arachidonic acid was neuronal (not glial) in origin and exhibited M1 muscarinic receptor pharmacology.
ESTHER : Kanterman_1990_Neurosci.Lett_118_235
PubMedSearch : Kanterman_1990_Neurosci.Lett_118_235
PubMedID: 2125713

Title : Carbachol-induced reverse transformation of Chinese hamster ovary cells transfected with and expressing the m5 muscarinic acetylcholine receptor - Felder_1989_FEBS.Lett_245_75
Author(s) : Felder CC , Ma AL , Conklin BR
Ref : FEBS Letters , 245 :75 , 1989
Abstract : Reverse transformation was induced in Chinese hamster ovary (CHO) cells transfected with and stably expressing the m5 subtype of the muscarinic acetylcholine receptor when stimulated with the muscarinic agonist, carbachol. Atropine, a muscarinic antagonist, blocked the carbachol-stimulated reverse transformation. CHO cells not transfected with the muscarinic receptor did not change with added carbachol. PMA induced reverse transformation without increasing cAMP accumulation in CHO cells. Carbachol, prostaglandin E2, and cholecystokinin increased cAMP accumulation but only carbachol caused reverse transformation. Carbachol-stimulated cAMP accumulation occurred at a higher concentration (EC50 10 microM) than did carbachol-stimulated reverse transformation (EC50 63 nM). Muscarinic m5 acetylcholine receptor transfected into CHO cells can induce reverse transformation which may be independent of cAMP.
ESTHER : Felder_1989_FEBS.Lett_245_75
PubMedSearch : Felder_1989_FEBS.Lett_245_75
PubMedID: 2466702

Title : Poster: Carbachol stimulation causes inhibition of mitogenesis and cell elongation in CHO cells transfected with muscarinrc receptor genes -
Author(s) : Conklin BR , Brann MR , Buckley NJ , Bonner TI , Ma AL , Felder CC , Axelrod J
Ref : Trends in Pharmacological Sciences , Suppl :117 , 1989

Title : A transfected m1 muscarinic acetylcholine receptor stimulates adenylate cyclase via phosphatidylinositol hydrolysis - Felder_1989_J.Biol.Chem_264_20356
Author(s) : Felder CC , Kanterman RY , Ma AL , Axelrod J
Ref : Journal of Biological Chemistry , 264 :20356 , 1989
Abstract : The m1 muscarinic acetylcholine receptor gene was transfected into and stably expressed in A9 L cells. The muscarinic receptor agonist, carbachol, stimulated inositol phosphate generation, arachidonic acid release, and cAMP accumulation in these cells. Carbachol stimulated arachidonic acid and inositol phosphate release with similar potencies, while cAMP generation required a higher concentration. Studies were performed to determine if the carbachol-stimulated cAMP accumulation was due to direct coupling of the m1 muscarinic receptor to adenylate cyclase via a GTP binding protein or mediated by other second messengers. Carbachol failed to stimulate adenylate cyclase activity in A9 L cell membranes, whereas prostaglandin E2 did, suggesting indirect stimulation. The phorbol ester, phorbol 12-myristate 13-acetate (PMA), stimulated arachidonic acid release yet inhibited cAMP accumulation in response to carbachol. PMA also inhibited inositol phosphate release in response to carbachol, suggesting that activation of phospholipase C might be involved in cAMP accumulation. PMA did not inhibit prostaglandin E2-, cholera toxin-, or forskolin-stimulated cAMP accumulation. The phospholipase A2 inhibitor eicosatetraenoic acid and the cyclooxygenase inhibitors indomethacin and naproxen had no effect on carbachol-stimulated cAMP accumulation. Carbachol-stimulated cAMP accumulation was inhibited with TMB-8, an inhibitor of intracellular calcium release, and W7, a calmodulin antagonist. These observations suggest that carbachol-stimulated cAMP accumulation does not occur through direct m1 muscarinic receptor coupling or through the release of arachidonic acid and its metabolites, but is mediated through the activation of phospholipase C. The generation of cytosolic calcium via inositol 1,4,5-trisphosphate and subsequent activation of calmodulin by m1 muscarinic receptor stimulation of phospholipase C appears to generate the accumulation of cAMP.
ESTHER : Felder_1989_J.Biol.Chem_264_20356
PubMedSearch : Felder_1989_J.Biol.Chem_264_20356
PubMedID: 2555356