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Gazzi_pyridine CB2R probes.pdf (1.17 MB)
Drug Derived Fluorescent Probes for the Specific Visualization of Cannabinoid Type 2 Receptor - A Toolbox Approach
Preprints are manuscripts made publicly available before they have been submitted for formal peer review and publication. They might contain new research findings or data. Preprints can be a draft or final version of an author's research but must not have been accepted for publication at the time of submission.
submitted on 12.11.2019, 08:54 and posted on 22.11.2019, 01:43by Gazzi, Thais, Benjamin Brennecke, Kenneth Atz, Claudia Korn, David Sykes, Roman C. Sarott, Matthias V. Westphal, Patrick Pfaff, Marie Weise, Yelena Mostinski, Bradley L. Hoare, Tamara Miljuš, Maira Mexi, Wolfgang Guba, André Anker, Arne C. Rufer, Eric A. Kusznir, Sylwia Huber, Catarina Raposo, Elisabeth A. Zirwes, Anja Osterwald, Anto Pavlovic, Svenja Moes, Jennifer Beck, Irene Benito-Cuesta, Teresa Grande, Faye Drawnel, Gabriella Widmer, Daniela Holzer, Tom van der Wel, Harpreet Mandhair, Yurii Saroz, Natasha Grimsey, Michael Honer, Jürgen Fingerle, Klaus Gawrisch, Julian Romero, Cecilia J. Hillard, Peter J. McCormick, Zoltan V. Varga, Mario van der Stelt, Pal Pacher, Jürg Gertsch, Christoph Ullmer, Sergio Oddi, Mauro Maccarrone, Dmitry B. Veprintsev, Erick M. Carreira, Uwe Grether, Marc Nazare
Cannabinoid type 2 receptor (CB2R) is
a fundamental part of the endocannabinoid signaling system (eCB system), and is
known to play an important role in tissue injury, inflammation, cancer and
pain. In stark contrast to its significance, the underlying signaling
mechanisms and tissue expression profiles are poorly understood. Due to its low
expression in healthy tissue and lack of reliable chemical tools, CB2R
visualization in live cells remains uncharted. Here we report the development
of a drug derived toolbox of highly potent, CB2R-selective
fluorescent probes based on reverse design. Extensive validation in several
applications such as CB2R detection in flow cytometry and
time-resolved imaging, and the development of a novel fluorescent-based TR-FRET
assay to generate kinetic and equilibrium binding data demonstrate the high
versatility of our toolbox. These probes are the first to preserve affinity and
efficacy in both human and mouse CB2R, a crucial aspect for
preclinical translatability, and to enable imaging of CB2R internalization in
living cells using confocal microscopy.