title: Generation and screening of oxime libraries addressing the neuronal GABA transporter GAT1.
abstract: The objective of the present study was to transfer the concept of library screening by MS binding assays, so far applied to pseudostatic hydrazine libraries, to static oxime libraries to screen for new potent inhibitors of mGAT1, the most abundant GABA transporter in the central nervous system that represents a validated drug target for the treatment of epilepsy. Library generation was performed by reaction of guvacine derivatives possessing a hydroxylamine functionality with various sets of four aldehydes. After dilution, the libraries were screened by competitive MS binding assays. Deconvolution experiments allowed hits in the most active libraries to be identified, and they were resynthesized for biological evaluation. That way a series of compounds was identified that displayed binding affinities ≥8.00 (pKi ) at mGAT1, one of which was found to be the most potent mGAT1 inhibitor known to date in a functional GABA uptake assay with a pIC50 value of 8.27 ± 0.03.
authors.affiliation: Department of Pharmacy, Center for Drug Research, Ludwig Maximilians University of Munich, Butenandtstrasse 5-13, 81377 Munich (Germany).
authors.forename: Felix T
authors.initials: FT
authors.lastname: Kern
authors.forename: Klaus T
authors.initials: KT
authors.lastname: Wanner
mesh_terms: D020650:Combinatorial Chemistry Techniques; D050485:GABA Plasma Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D058805:GABA Uptake Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D010091:Oximes / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D013329:Structure-Activity Relationship; D005680:gamma-Aminobutyric Acid / Q000737:chemistry / Q000378:metabolism
chemical_list: D050485:GABA Plasma Membrane Transport Proteins; D058805:GABA Uptake Inhibitors; D010091:Oximes; D005680:gamma-Aminobutyric Acid
keywords: binding assays; dynamic combinatorial chemistry; mass spectrometry; membrane proteins; oximes