Abstract
The recently identified mas-related-gene (MRG) family of receptors, located primarily in sensory neurons of the dorsal root ganglion, has been implicated in the perception of pain. Thus, antagonists of this class of receptors have been postulated to be useful analgesics. Toward this end, we developed a cell-based beta-lactamase (BLA) reporter gene assay to identify small molecule antagonists of the human MRG-X1 receptor from a library of compounds. Single-cell clones expressing functional receptors were selected using the BLA reporter gene technology. The EC50 for the MRG agonist peptide, BAM15, appeared to be comparable between the BLA assay and the intracellular Ca2+ transient assays in these cells. Ultra high-throughput screening of approximately 1 million compounds in a 1.8-microl cell-based BLA reporter gene assay was conducted in a 3456-well plate format. Compounds exhibiting potential antagonist profile in the BLA assay were confirmed in the second messenger Ca2+ transient assay. A cell-based receptor trafficking assay was used to further validate the mechanism of action of these compounds. Several classes of compounds, particularly the 2,3-disubstituted azabicyclo-octanes, appear to be relatively potent antagonists at t...Continue Reading
References
Jul 1, 1983·Peptides·R Quirion, A S Weiss
Feb 1, 1994·Journal of Neuroendocrinology·C J BoersmaF W Van Leeuwen
May 20, 1993·Nature·B R ConklinH R Bourne
Oct 3, 1996·Nature·O B GoodmanJ L Benovic
Jan 24, 1998·Science·G ZlokarnikR Y Tsien
Aug 27, 1999·Trends in Pharmacological Sciences·A MarcheseB F O'Dowd
Mar 17, 2000·Science·J Drews
Mar 16, 2001·Trends in Neurosciences·O CivelliR K Reinscheid
Sep 12, 2001·Cell·X DongD J Anderson
Feb 19, 2002·Nature Neuroscience·Paola M C LemboSultan Ahmad
Sep 5, 2002·Nature Reviews. Drug Discovery·Andrew L Hopkins, Colin R Groom
Feb 28, 2003·Analytical Biochemistry·Peter HodderBerta Strulovici
May 17, 2003·Nature Reviews. Drug Discovery·Konrad H BleicherAlexander I Alanine
Apr 20, 2004·Assay and Drug Development Technologies·Robert H OakleyCarson R Loomis
May 14, 2004·Journal of Biomolecular Screening·Oleg KornienkoBerta Strulovici
Aug 7, 2004·Journal of Biomolecular Screening·Peter HodderBerta Strulovici
Citations
Sep 14, 2011·Histochemistry and Cell Biology·Leela R AvulaJean-Pierre Timmermans
Mar 19, 2013·Histochemistry and Cell Biology·Leela Rani AvulaJean-Pierre Timmermans
Oct 12, 2010·Journal of Biomedicine & Biotechnology·Seena K AjitEdward J Kaftan
Mar 19, 2013·PloS One·Hans Jürgen SolinskiAndreas Breit
Dec 1, 2015·European Journal of Pharmacology·Wolfgang SchröderJuan Fernando Herrero
Oct 19, 2006·Current Opinion in Biotechnology·Moo-Yeal Lee, Jonathan S Dordick
Dec 18, 2013·Pain·Shao-Qiu HeYun Guan
May 29, 2014·Pharmacological Reviews·Hans Jürgen SolinskiAndreas Breit
May 23, 2007·Journal of Biomolecular Screening·Tina M HallisTammy Turek-Etienne
Feb 24, 2019·Stem Cell Research & Therapy·Mingjun TangJian Ge
Sep 24, 2014·Pharmacological Reviews·Michael BaderRobson A Santos
Feb 23, 2017·Proceedings of the National Academy of Sciences of the United States of America·Zhe LiXinzhong Dong
Jul 1, 2019·Itch·Pang-Yen TsengXinzhong Dong
Apr 1, 2021·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Hayley McMillanIkhlas El Karim
Jan 17, 2009·Journal of Medicinal Chemistry·Berthold WroblowskiWilliam J Coates