title: Evidence for the presence of three distinct binding sites for the thioflavin T class of Alzheimer's disease PET imaging agents on beta-amyloid peptide fibrils.
abstract: Imaging the progression of Alzheimer's disease would greatly facilitate the discovery of therapeutics, and a wide range of ligands are currently under development for the detection of beta-amyloid peptide (Abeta)-containing plaques by using positron emission tomography. Here we report an in-depth characterization of the binding of seven previously described ligands to in vitro generated Abeta-(1-40) polymers. All of the compounds were derived from the benzothiazole compound thioflavin T and include 2-[4'-(methylamino)phenyl]benzothiazole and 2-(4'-dimethylamino-)phenyl-imidazo[1,2-a]-pyridine derivatives, 2-[4'-(dimethylamino)phenyl]-6-iodobenzothiazole and 2-[4'-(4''-methylpiperazin-1-yl)phenyl]-6-iodobenzothiazole, and a benzofuran compound (5-bromo-2-(4-dimethylaminophenyl)benzofuran). By using a range of fluorescent and radioligand binding assays, we find that these compounds display a more complex binding pattern than described previously and are consistent with three classes of binding sites on the Abeta fibrils. All of the compounds bound with very high affinity (low nm K(d)) to a low capacity site (BS3) (1 ligand-binding site per approximately 300 Abeta-(1-40) monomers) consistent with the previously recognized binding site for these compounds on the fibrils. However, the compounds also bound with high affinity (K(d) approximately 100 nm) to either one of two additional binding sites on the Abeta-(1-40) polymer. The properties of these sites, BS1 and BS2, suggest they are adjacent or partially overlapping and have a higher capacity than BS3, occurring every approximately 35 or every approximately 4 monomers of Abeta-(1-40)-peptide, respectively. Compounds appear to display selectivity for BS2 based on the presence of a halogen substitution (2-[4'-(dimethylamino)phenyl]-6-iodobenzothiazole, 2-[4'-(4''-methylpiperazin-1-yl)phenyl]-6-iodobenzothiazole, and 5-bromo-2-(4-dimethylaminophenyl)benzofuran) on their aromatic ring system. The presence of additional ligand-binding sites presents potential new targets for ligand development and may allow a more complete modeling of the current positron emission tomography data.
authors.affiliation: Translational Medicine and Technology, GlaxoSmithKline Research and Development, Addenbrookes Hospital, Cambridge CB2 2GG, United Kingdom. andrew.2.lockhart@gsk.com
authors.forename: Andrew
authors.initials: A
authors.lastname: Lockhart
authors.forename: Liang
authors.initials: L
authors.lastname: Ye
authors.forename: Duncan B
authors.initials: DB
authors.lastname: Judd
authors.forename: Andy T
authors.initials: AT
authors.lastname: Merritt
authors.forename: Peter N
authors.initials: PN
authors.lastname: Lowe
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Morgenstern
authors.forename: Guizhu
authors.initials: G
authors.lastname: Hong
authors.forename: Antony D
authors.initials: AD
authors.lastname: Gee
authors.forename: John
authors.initials: J
authors.lastname: Brown
mesh_terms: D000544:Alzheimer Disease / Q000473:pathology*; D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000378:metabolism; D052160:Benzothiazoles; D001665:Binding Sites; D001667:Binding, Competitive; D002499:Centrifugation, Density Gradient; D004305:Dose-Response Relationship, Drug; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D007700:Kinetics; D008024:Ligands; D008954:Models, Biological; D008956:Models, Chemical; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D049268:Positron-Emission Tomography / Q000379:methods*; D011485:Protein Binding; D013050:Spectrometry, Fluorescence; D013844:Thiazoles / Q000737:chemistry*
chemical_list: D016229:Amyloid beta-Peptides; D052160:Benzothiazoles; D008024:Ligands; D010446:Peptide Fragments; D010455:Peptides; D013844:Thiazoles; C078592:amyloid beta-protein (1-40); C009462:thioflavin T