title: In vitro pharmacological and rat pharmacokinetic characterization of LY3020371, a potent and selective mGlu2/3receptor antagonist.
abstract: Metabotropic glutamate2/3(mGlu2/3) receptors are of considerable interest owing to their role in modulating glutamate transmission via presynaptic, postsynaptic and glial mechanisms. As part of our ongoing efforts to identify novel ligands for these receptors, we have discovered (1S,2R,3S,4S,5R,6R)-2-amino-3-[(3,4-difluorophenyl)sulfanylmethyl]-4-hydroxy-bicyclo[3.1.0]hexane-2,6-dicarboxylic acid; (LY3020371), a potent and selective orthosteric mGlu2/3receptor antagonist. In this account, we characterize the effects of LY3020371 in membranes and cells expressing human recombinant mGlu receptor subtypes as well as in native rodent and human brain tissue preparations, providing important translational information for this molecule. In membranes from cells expressing recombinant human mGlu2and mGlu3receptor subtypes, LY3020371.HCl competitively displaced binding of the mGlu2/3agonist ligand [3H]-459477 with high affinity (hmGlu2Ki= 5.26 nM; hmGlu3Ki = 2.50 nM). In cells expressing hmGlu2receptors, LY3020371.HCl potently blocked mGlu2/3agonist (DCG-IV)-inhibited, forskolin-stimulated cAMP formation (IC50= 16.2 nM), an effect that was similarly observed in hmGlu3-expressing cells (IC50= 6.21 nM). Evaluation of LY3020371 in cells expressing the other human mGlu receptor subtypes revealed high mGlu2/3receptor selectivity. In rat native tissue assays, LY3020371 demonstrated effective displacement of [3H]-459477 from frontal cortical membranes (Ki= 33 nM), and functional antagonist activity in cortical synaptosomes measuring both the reversal of agonist-suppressed second messenger production (IC50= 29 nM) and agonist-inhibited, K+-evoked glutamate release (IC50= 86 nM). Antagonism was fully recapitulated in both primary cultured cortical neurons where LY3020371 blocked agonist-suppressed spontaneous Ca2+oscillations (IC50= 34 nM) and in an intact hippocampal slice preparation (IC50= 46 nM). Functional antagonist activity was similarly demonstrated in synaptosomes prepared from epileptic human cortical or hippocampal tissues, suggesting a translation of the mGlu2/3antagonist pharmacology from rat to human. Intravenous dosing of LY3020371 in rats led to cerebrospinal fluid drug levels that are expected to effectively block mGlu2/3receptors in vivo. Taken together, these results establish LY3020371 as an important new pharmacological tool for studying mGlu2/3receptors in vitro and in vivo. This article is part of the Special Issue entitled 'Metabotropic Glutamate Receptors, 5 years on'.
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Witkin
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
authors.forename: Paul L
authors.initials: PL
authors.lastname: Ornstein
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
authors.forename: Charles H
authors.initials: CH
authors.lastname: Mitch
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
authors.forename: Renhua
authors.initials: R
authors.lastname: Li
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
authors.forename: Stephon C
authors.initials: SC
authors.lastname: Smith
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
authors.forename: Beverly A
authors.initials: BA
authors.lastname: Heinz
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
authors.forename: Xu-Shan
authors.initials: XS
authors.lastname: Wang
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
authors.forename: Chuanxi
authors.initials: C
authors.lastname: Xiang
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
authors.forename: Joan H
authors.initials: JH
authors.lastname: Carter
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
authors.forename: Wesley H
authors.initials: WH
authors.lastname: Anderson
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
authors.forename: Xia
authors.initials: X
authors.lastname: Li
authors.affiliation: Windlesham, Surrey, UK.
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Broad
authors.affiliation: Windlesham, Surrey, UK.
authors.forename: Francesca
authors.initials: F
authors.lastname: Pasqui
authors.affiliation: Windlesham, Surrey, UK.
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Fitzjohn
authors.affiliation: Windlesham, Surrey, UK.
authors.forename: Helen E
authors.initials: HE
authors.lastname: Sanger
authors.affiliation: Indiana University/Purdue University, USA.
authors.forename: Jodi L
authors.initials: JL
authors.lastname: Smith
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
authors.forename: John
authors.initials: J
authors.lastname: Catlow
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
authors.forename: Steven
authors.initials: S
authors.lastname: Swanson
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA. Electronic address: monn@lilly.com.
authors.forename: James A
authors.initials: JA
authors.lastname: Monn
mesh_terms: D000818:Animals; D002460:Cell Line; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism; D003510:Cyclohexanes / Q000737:chemistry / Q000493:pharmacokinetics*; D004305:Dose-Response Relationship, Drug; D018691:Excitatory Amino Acid Antagonists / Q000737:chemistry / Q000493:pharmacokinetics*; D006801:Humans; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D018094:Receptors, Metabotropic Glutamate / Q000037:antagonists & inhibitors* / Q000378:metabolism*
chemical_list: C000620392:2-amino-3-((3,4-difluorophenyl)sulfanylmethyl)-4-hydroxy-bicyclo(3.1.0)hexane-2,6-dicarboxylic acid hydrochloride; D003510:Cyclohexanes; D018691:Excitatory Amino Acid Antagonists; D018094:Receptors, Metabotropic Glutamate; C108822:metabotropic glutamate receptor 2; C102718:metabotropic glutamate receptor 3
keywords: LY3020371; Metabotropic glutamate receptors; mGlu(2); mGlu(3)