title: Stereochemistry and molecular pharmacology of (S)-thio-ATPA, a new potent and selective GluR5 agonist.
abstract: (RS)-2-Amino-3-(5-tert-butyl-3-hydroxy-4-isothiazolyl)propionic acid (thio-ATPA), a 3-isothiazolol analogue of (RS)-2-amino-3-(5-tert-butyl-3-hydroxy-4-isoxazolyl)propionic acid (ATPA), has previously been shown to be a relatively weak AMPA receptor agonist at native (S)-glutamic acid ((S)-Glu) receptors (EC(50)=14 microM), comparable in potency with ATPA (EC(50)=34 microM). Recent findings, that (S)-ATPA is a potent (EC(50)=0.48 microM) and selective agonist at homomerically expressed ionotropic GluR5, prompted us to resolve thio-ATPA using chiral chromatography and pharmacologically characterize the two enantiomers at native as well as cloned ionotropic glutamate receptors. The enantiomers, (S)- and (R)-thio-ATPA, were obtained in high enantiomeric excess, and their absolute stereochemistry established by an X-ray crystallographic analysis. Electrophysiologically, the two enantiomers were evaluated in the rat cortical wedge preparation, and the S-enantiomer was found to be an AMPA receptor agonist (EC(50)=8.7 microM) twice as potent as the racemate, whereas the R-enantiomer was devoid of activity. In accordance with this, (S)-thio-ATPA proved to be an agonist at homomerically expressed recombinant AMPA receptors (GluR1o, GluR3o, and GluR4o) with EC(50) values of 5, 32 and 20 microM, respectively, producing maximal steady state currents of 78--168% of those maximally evoked by kainic acid, and 120-1600% of those maximally evoked by (S)-ATPA. At homomerically expressed GluR5, (S)-thio-ATPA was found to be a potent agonist (EC(50)=0.10 microM), thus being approximately five times more potent than (S)-ATPA. (R)-Thio-ATPA induced saturating currents with an estimated EC(50) value of 10 microM, most likely due to a contamination with (S)-thio-ATPA. At heteromerically expressed GluR6+KA2 receptors, (S)-thio-ATPA showed relatively weak agonistic properties (EC(50)=4.9 microM). Thus, (S)-thio-ATPA has been shown to be a very potent agonist at GluR5, and may be a valuable tool for the investigation of desensitization properties of AMPA receptors.
authors.affiliation: NeuroScience PharmaBiotec Research Center, Department of Medicinal Chemistry, The Royal Danish School of Pharmacy, 2 Universitetsparken, DK-2100 Copenhagen, Denmark.
authors.forename: T B
authors.initials: TB
authors.lastname: Stensbøl
authors.forename: H S
authors.initials: HS
authors.lastname: Jensen
authors.forename: B
authors.initials: B
authors.lastname: Nielsen
authors.forename: T N
authors.initials: TN
authors.lastname: Johansen
authors.forename: J
authors.initials: J
authors.lastname: Egebjerg
authors.forename: K
authors.initials: K
authors.lastname: Frydenvang
authors.forename: P
authors.initials: P
authors.lastname: Krogsgaard-Larsen
mesh_terms: D000409:Alanine / Q000031:analogs & derivatives / Q000737:chemistry* / Q000494:pharmacology*; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D018690:Excitatory Amino Acid Agonists / Q000494:pharmacology*; D066298:In Vitro Techniques; D007700:Kinetics; D008564:Membrane Potentials / Q000187:drug effects; D008968:Molecular Conformation; D009865:Oocytes / Q000378:metabolism; D018408:Patch-Clamp Techniques; D051381:Rats; D018091:Receptors, AMPA / Q000819:agonists; D013237:Stereoisomerism; D013844:Thiazoles / Q000737:chemistry* / Q000494:pharmacology*; D014158:Transcription, Genetic; D014982:Xenopus laevis
chemical_list: C428244:2-amino-3-(5-tert-butyl-3-hydroxy-4-isothiazolyl)propionic acid; D018690:Excitatory Amino Acid Agonists; D018091:Receptors, AMPA; D013844:Thiazoles; D000409:Alanine