High-Throughput Screening for Allosteric Modulators of GPCRs

2015 ◽  
pp. 223-240 ◽  
Author(s):  
Robert L. Bertekap ◽  
Neil T. Burford ◽  
Zhuyin Li ◽  
Andrew Alt
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Allosteric Modulators

scholarly journals Discovery of Selective Cannabinoid CB2 Receptor Agonists by High-Throughput Screening

2017 ◽  
Vol 23 (4) ◽  
pp. 375-383 ◽  
Author(s):  
Lisa M. Ogawa ◽  
Neil T. Burford ◽  
Yu-Hsien Liao ◽  
Caitlin E. Scott ◽  
Ashley M. Hine ◽  
...  
Keyword(s):  
Side Effects ◽  
High Throughput ◽  
High Throughput Screening ◽  
Immune Modulation ◽  
Cannabinoid Receptors ◽  
Endocannabinoid System ◽  
Allosteric Modulators ◽  
Peripheral Tissues ◽  
Selective Agonists

The endocannabinoid system (ECS) plays a diverse role in human physiology ranging from the regulation of mood and appetite to immune modulation and the response to pain. Drug development that targets the cannabinoid receptors (CB1 and CB2) has been explored; however, success in the clinic has been limited by the psychoactive side effects associated with modulation of the neuronally expressed CB1 that are enriched in the CNS. CB2, however, are expressed in peripheral tissues, primarily in immune cells, and thus development of CB2-selective drugs holds the potential to modulate pain among other indications without eliciting anxiety and other undesirable side effects associated with CB1 activation. As part of a collaborative effort among industry and academic laboratories, we performed a high-throughput screen designed to discover selective agonists or positive allosteric modulators (PAMs) of CB2. Although no CB2 PAMs were identified, 167 CB2 agonists were discovered here, and further characterization of four select compounds revealed two with high selectivity for CB2 versus CB1. These results broaden drug discovery efforts aimed at the ECS and may lead to the development of novel therapies for immune modulation and pain management with improved side effect profiles.

scholarly journals High-Throughput Screening for Positive Allosteric Modulators Identified Potential Therapeutics against Acetylcholinesterase Inhibition

2015 ◽  
Vol 20 (9) ◽  
pp. 1142-1149 ◽  
Author(s):  
Richard R. Chapleau ◽  
Craig A. McElroy ◽  
Christopher D. Ruark ◽  
Emily J. Fleming ◽  
Amy B. Ghering ◽  
...  
Keyword(s):  
Side Effects ◽  
High Throughput ◽  
High Throughput Screening ◽  
Standard Of Care ◽  
Acetylcholinesterase Inhibition ◽  
Therapeutic Window ◽  
Ache Inhibitor ◽  
Allosteric Modulators ◽  
Current Standard ◽  
Positive Allosteric Modulators

The current standard of care for treatment of organophosphate (OP) poisoning includes pretreatment with the weak reversible acetylcholinesterase (AChE) inhibitor pyridostigmine bromide. Because this drug is an AChE inhibitor, similar side effects exist as with OP poisoning. In an attempt to provide a therapeutic capable of mitigating AChE inhibition without such side effects, high-throughput screening was performed to identify a compound capable of increasing the catalytic activity of AChE. Herein, two such novel positive allosteric modulators (PAMs) of AChE are presented. These PAMs increase AChE activity threefold, but they fail to upshift the apparent IC50 of a variety of OPs. Further development and optimization of these compounds may lead to pre- and/or postexposure therapeutics with broad-spectrum efficacy against pesticide and nerve agent poisoning. In addition, they could be used to complement the current therapeutic standard of care to increase the activity of uninhibited AChE, potentially increasing the efficacy of current therapeutics in addition to altering the therapeutic window.

scholarly journals An Integrated Approach for Screening and Identification of Positive Allosteric Modulators of N-Methyl-D-Aspartate Receptors

2016 ◽  
Vol 21 (5) ◽  
pp. 468-479 ◽  
Author(s):  
Enrique Jambrina ◽  
Rok Cerne ◽  
Emery Smith ◽  
Louis Scampavia ◽  
Maria Cuadrado ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Therapeutic Interventions ◽  
Calcium Flux ◽  
Electrophysiological Recording ◽  
Small Subset ◽  
Allosteric Modulators ◽  
Chemical Library ◽  
Subtype Selectivity ◽  
Positive Allosteric Modulators

N-methyl-D-aspartate receptors (NMDARs) are ionotropic glutamate receptors that play an important role in synaptic plasticity and learning and memory formation. Malfunctioning of NMDARs, in particular the reduction in NMDAR activity, is thought to be implicated in major neurological disorders. NMDAR positive allosteric modulators (PAMs) represent potential therapeutic interventions for restoring normal NMDAR function. We report a novel screening approach for identification and characterization of NMDAR-PAMs. The approach combines high-throughput fluorescence imaging with automated electrophysiological recording of glutamate-evoked responses in HEK-293 cells expressing NR1/NR2A NMDAR subunits. Initial high-throughput screening (HTS) of a chemical library containing >810,000 compounds using a calcium flux assay in 1536-well plate format identified a total of 864 NMDAR-PAMs. Concentration response determination in both calcium flux and automated electrophysiological assays found several novel chemical series with EC50 values between 0.49 and 10 µM. A small subset (six series) was selected and analyzed for pharmacological properties, subtype selectivity, mode of action, and activity at native NMDARs. Our approach demonstrates the successful application of HTS functional assays that led to identification of NMDAR-PAMs providing the foundation for further medicinal chemistry work that may lead to novel therapies for treatment of cognitive impairment associated with Alzheimer’s disease and schizophrenia.

ChemInform Abstract: Synthesis and SAR of Novel, 4-(Phenylsulfamoyl)phenylacetamide mGlu4 Positive Allosteric Modulators (PAMs) Identified by Functional High-Throughput Screening (HTS).

ChemInform ◽  
2010 ◽  
Vol 42 (1) ◽  
pp. no-no
Author(s):  
Darren W. Engers ◽  
Patrick R. Gentry ◽  
Richard Williams ◽  
Julie D. Bolinger ◽  
C. David Weaver ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Allosteric Modulators ◽  
Positive Allosteric Modulators

New positive allosteric modulators of the metabotropic glutamate receptor 2 (mGluR2). Identification and synthesis of N-propyl-5-substituted isoquinolones

MedChemComm ◽  
2011 ◽  
Vol 2 (2) ◽  
pp. 132-139 ◽  
Author(s):  
Andrés A. Trabanco ◽  
Guillaume Duvey ◽  
José María Cid ◽  
Gregor J. Macdonald ◽  
Philippe Cluzeau ◽  
...  
Keyword(s):  
Glutamate Receptor ◽  
High Throughput ◽  
High Throughput Screening ◽  
Metabotropic Glutamate Receptor ◽  
Allosteric Modulators ◽  
Metabotropic Glutamate ◽  
Positive Allosteric Modulators

N-propyl-5-substituted isoquinolones were identified as mGluR2 PAMs via high-throughput screening (HTS). Initial SAR exploration led to the identification of compound 20.

scholarly journals High‐throughput screening for novel allosteric modulators of the D1 dopamine receptor (662.4)

2014 ◽  
Vol 28 (S1) ◽  
Author(s):  
Jennie Conroy ◽  
R. Free ◽  
Trevor Doyle ◽  
Noel Southall ◽  
Lisa Hazelwood ◽  
...  
Keyword(s):  
Dopamine Receptor ◽  
High Throughput ◽  
High Throughput Screening ◽  
Allosteric Modulators ◽  
D1 Dopamine Receptor

scholarly journals High‐throughput screening for identification of novel allosteric modulators of the D 3 dopamine receptor

2015 ◽  
Vol 29 (S1) ◽  
Author(s):  
A Moritz ◽  
R. Free ◽  
W. Weiner ◽  
M. Bachani ◽  
J. Conroy ◽  
...  
Keyword(s):  
Dopamine Receptor ◽  
High Throughput ◽  
High Throughput Screening ◽  
Allosteric Modulators

scholarly journals Use of FLIPR Membrane Potential Dyes for Validation of High-Throughput Screening with the FLIPR and µARCS Technologies: Identification of Ion Channel Modulators Acting on the GABAA Receptor

2008 ◽  
Vol 13 (3) ◽  
pp. 218-228 ◽  
Author(s):  
Christoph Joesch ◽  
Emelie Guevarra ◽  
Serge P. Parel ◽  
Andreas Bergner ◽  
Peter Zbinden ◽  
...  
Keyword(s):  
Membrane Potential ◽  
High Throughput ◽  
High Throughput Screening ◽  
Constitutive Expression ◽  
Initial Assessment ◽  
Blue Dye ◽  
Imaging Plate ◽  
Allosteric Modulators ◽  
Positive Allosteric Modulators ◽  
Red Dye

Fluorometric imaging plate reader (FLIPR) membrane potential dyes (FMP-Red-Dye and FMP-Blue-Dye) were evaluated for the detection of compounds acting either as positive allosteric modulators or agonists on the GABAA receptor (GABAAR). A stable HEK293 cell line with constitutive expression of the rat GABA AR α1, β2, and γ2 genes was used to establish a functional high-throughput screening (HTS) assay based on measurement of the membrane potential change in living cells. The assay was validated with the FLIPR technology for identification of agonists and positive allosteric modulators using GABA and diazepam as model compounds. The FMP-Red-Dye showed better performance than the FMP-Blue-Dye, and the effects induced by GABA and diazepam were comparable to electrophysiology data. Subsequently, the assay was also validated with an ultra-HTS approach known as microarrayed compound screening (µARCS). The LOPAC library was used in a test screen for an initial assessment of the technology. Finally, the FLIPR and µARCS technologies were tested with a larger screening campaign. A focused library of 3520 putative positive modulators was tested with the FLIPR assay, and a diverse subset of 84,480 compounds was selected for screening with the µARCS technology. All hits were subjected to verification using the FLIPR technology, and confirmed hits were subsequently evaluated by EC50 determination. Finally, selected hits were further confirmed with electrophysiology testing. ( Journal of Biomolecular Screening 2008:218-228)

scholarly journals Synthesis and SAR of novel, 4-(phenylsulfamoyl)phenylacetamide mGlu4 positive allosteric modulators (PAMs) identified by functional high-throughput screening (HTS)

2010 ◽  
Vol 20 (17) ◽  
pp. 5175-5178 ◽  
Author(s):  
Darren W. Engers ◽  
Patrick R. Gentry ◽  
Richard Williams ◽  
Julie D. Bolinger ◽  
C. David Weaver ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Allosteric Modulators ◽  
Positive Allosteric Modulators
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