title: Azidobupramine, an Antidepressant-Derived Bifunctional Neurotransmitter Transporter Ligand Allowing Covalent Labeling and Attachment of Fluorophores.
abstract: The aim of this study was to design, synthesize and validate a multifunctional antidepressant probe that is modified at two distinct positions. The purpose of these modifications was to allow covalent linkage of the probe to interaction partners, and decoration of probe-target complexes with fluorescent reporter molecules. The strategy for the design of such a probe (i.e., azidobupramine) was guided by the need for the introduction of additional functional groups, conveying the required properties while keeping the additional moieties as small as possible. This should minimize the risk of changing antidepressant-like properties of the new probe azidobupramine. To control for this, we evaluated the binding parameters of azidobupramine to known target sites such as the transporters for serotonin (SERT), norepinephrine (NET), and dopamine (DAT). The binding affinities of azidobupramine to SERT, NET, and DAT were in the range of structurally related and clinically active antidepressants. Furthermore, we successfully visualized azidobupramine-SERT complexes not only in SERT-enriched protein material but also in living cells stably overexpressing SERT. To our knowledge, azidobupramine is the first structural analogue of a tricyclic antidepressant that can be covalently linked to target structures and further attached to reporter molecules while preserving antidepressant-like properties and avoiding radioactive isotopes.
authors: Max Planck Institute of Psychiatry, Clinical Department, Munich, Germany.
authors: Thomas
authors: T
authors: Kirmeier
authors: Max Planck Institute of Psychiatry, Department of Translational Research in Psychiatry, Munich, Germany.
authors: Ranganath
authors: R
authors: Gopalakrishnan
authors: Max Planck Institute of Psychiatry, Department of Translational Research in Psychiatry, Munich, Germany.
authors: Vanessa
authors: V
authors: Gormanns
authors: Max Planck Institute of Psychiatry, Department of Translational Research in Psychiatry, Munich, Germany.
authors: Anna M
authors: AM
authors: Werner
authors: Max Planck Institute of Psychiatry, Department of Translational Research in Psychiatry, Munich, Germany.
authors: Serena
authors: S
authors: Cuboni
authors: Technical University Munich, IAS, CIPSM, Department of Chemistry, Garching, Germany.
authors: Georg C
authors: GC
authors: Rudolf
authors: Department Pharmazie Zentrum für Pharmaforschung, Ludwig-Maximilians-Universität München, Munich, Germany.
authors: Georg
authors: G
authors: Höfner
authors: Department Pharmazie Zentrum für Pharmaforschung, Ludwig-Maximilians-Universität München, Munich, Germany.
authors: Klaus T
authors: KT
authors: Wanner
authors: Technical University Munich, IAS, CIPSM, Department of Chemistry, Garching, Germany.
authors: Stephan A
authors: SA
authors: Sieber
authors: Max Planck Institute of Psychiatry, Department of Translational Research in Psychiatry, Munich, Germany.
authors: Ulrike
authors: U
authors: Schmidt
authors: Max Planck Institute of Psychiatry, Clinical Department, Munich, Germany.
authors: Florian
authors: F
authors: Holsboer
authors: Max Planck Institute of Psychiatry, Department of Translational Research in Psychiatry, Munich, Germany.
authors: Theo
authors: T
authors: Rein
authors: Max Planck Institute of Psychiatry, Department of Translational Research in Psychiatry, Munich, Germany.
authors: Felix
authors: F
authors: Hausch
mesh_terms: D000588:Amines / Q000737:chemistry; D000929:Antidepressive Agents, Tricyclic / Q000138:chemical synthesis / Q000737:chemistry*; D001381:Azepines / Q000138:chemical synthesis / Q000737:chemistry*; D001665:Binding Sites; D002460:Cell Line; D004298:Dopamine / Q000378:metabolism; D050483:Dopamine Plasma Membrane Transport Proteins / Q000737:chemistry / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D015870:Gene Expression; D006801:Humans; D007700:Kinetics; D008024:Ligands; D015335:Molecular Probes / Q000138:chemical synthesis / Q000737:chemistry*; D009638:Norepinephrine / Q000378:metabolism; D050484:Norepinephrine Plasma Membrane Transport Proteins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D012701:Serotonin / Q000378:metabolism; D050486:Serotonin Plasma Membrane Transport Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000588:Amines; D000929:Antidepressive Agents, Tricyclic; D001381:Azepines; D050483:Dopamine Plasma Membrane Transport Proteins; D005456:Fluorescent Dyes; D008024:Ligands; D015335:Molecular Probes; D050484:Norepinephrine Plasma Membrane Transport Proteins; C493227:SLC6A2 protein, human; C493230:SLC6A4 protein, human; D050486:Serotonin Plasma Membrane Transport Proteins; D012701:Serotonin; D004298:Dopamine; D009638:Norepinephrine