Development of High-Specificity Fluorescent Probes to Enable Cannabinoid Type 2 Receptor Studies in Living Cells

Sarott, Roman C.; Westphal, Matthias V.; Pfaff, Patrick; Korn, Claudia; Sykes, David A.; Gazzi, Thais; Brennecke, Benjamin; Atz, Kenneth; Weise, Marie; Mostinski, Yelena; Hompluem, Pattarin; Koers, Eline J.; Miljuš, Tamara; Roth, Nicolas; Asmelash, Hermon; Vong, Man; Piovesan, Jacopo; Guba, Wolfgang; Rufer, Arne C.; Kusznir, Eric A.; Huber, Sylwia; Raposo, Catarina; Zirwes, Elisabeth A.; Osterwald, Anja; Pavlovic, Anto; Moes, Svenja; Beck, Jennifer; Benito-Cuesta, Irene; Grande, Teresa; Esteban, Samuel Ruiz de Martı N; Yeliseev, Alexei; Drawnel, Faye; Widmer, Gabriella; Holzer, Daniela; Wel, Tom van der; Mandhair, Harpreet; Yuan, Cheng-Yin; Drobyski, William R.; Saroz, Yurii; Grimsey, Natasha L.; Honer, Michael; Fingerle, Jürgen; Gawrisch, Klaus; Romero, Julián; Hillard, Cecilia J.; Varga, Zoltán V.; Stelt, Mario van der; Pacher, Pál; Gertsch, Jürg; McCormick, Peter J.; Ullmer, Christoph; Oddi, Sergio; Maccarrone, Mauro; Veprintsev, Dmitry B.; Nazaré, Marc; Grether, Uwe; Carreira, Erick M.

doi:10.1021/jacs.0c05587

Cited by 34 publications

(71 citation statements)

References 70 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Furthermore, they evaluated kinetic binding parameters in a novel TR‐FRET‐based assay, which is suitable for high‐throughput screening. Interestingly, the authors also synthesized a compound with an AttoThio12 dye, rendering it a partial agonist with high selectivity (K i CB 2 /CB 1 ratio=228), but lower efficacy [161] …”

Section: Fluorescent Ligands For Gpcrsmentioning

confidence: 99%

Lighting Up the Plasma Membrane: Development and Applications of Fluorescent Ligands for Transmembrane Proteins

Borgarelli

Klingl

Escamilla-Ayala

et al. 2021

Chemistry A European J

View full text Add to dashboard Cite

Despite the fact that transmembrane proteins represent the main therapeutic targets for decades, complete and in‐depth knowledge about their biochemical and pharmacological profiling is not fully available. In this regard, target‐tailored small‐molecule fluorescent ligands are a viable approach to fill in the missing pieces of the puzzle. Such tools, coupled with the ability of high‐precision optical techniques to image with an unprecedented resolution at a single‐molecule level, helped unraveling many of the conundrums related to plasma proteins’ life‐cycle and druggability. Herein, we review the recent progress made during the last two decades in fluorescent ligand design and potential applications in fluorescence microscopy of voltage‐gated ion channels, ligand‐gated ion channels and G‐coupled protein receptors.

show abstract

Section: Fluorescent Ligands For Gpcrsmentioning

confidence: 99%

Lighting Up the Plasma Membrane: Development and Applications of Fluorescent Ligands for Transmembrane Proteins

Borgarelli

Klingl

Escamilla-Ayala

et al. 2021

Chemistry A European J

View full text Add to dashboard Cite

show abstract

“…Further establishing upon the scaffold, the fluorophore was altered to DY-480XL, Alexa647, and AttoThio12, while the terminal azido remained consistent. [141,142] From these compounds, the DY-480XL (64) and AttoThio12 (65) dyes demonstrated high affinity and selectivity to CB 2 R. Both probes were suggested to be alternatives to radioligands as per a TR-FRET assay. Notably, 65 could be detected in 5xFAD mice, expressing mCB 2 R AD microglia, and labelled endogenous hCB 2 R MDA-MB-231 breast cancer cells by flow cytometry.…”

Section: Bifunctional Fluorescent Probesmentioning

confidence: 99%

Imaging Cannabinoid Receptors: A Brief Collection of Covalent and Fluorescent Probes for CB

Hamilton¹,

Payne²,

Mocerino³

et al. 2021

Aust. J. Chem.

View full text Add to dashboard Cite

There has been an expanding public interest towards the notion that modulation of the sophisticated endocannabinoid system can lead to various therapeutic benefits that are yet to be fully explored. In recent years, the drug discovery paradigm in this field has been largely based on the development of selective CB 2 receptor agonists, avoiding the unwanted CB 1 receptor-mediated psychoactive side effects. Mechanistically, target engagement studies are crucial for confirming the ligand-receptor interaction and the subsequent biological cascades that lead to the observed therapeutic effects. Concurrently, imaging techniques for visualisation of cannabinoid receptors are increasingly reported in the literature. Small molecule imaging tools ranging from phytocannabinoids such as tetrahydrocannabinol (THC) and cannabidiol (CBD) to the endocannabinoids as well as the purely synthetic cannabimimetics, have been explored to date with varying degrees of success. This Review will cover currently known photoactivatable, electrophilic, and fluorescent ligands for both the CB 1 and CB 2 receptors. Structural insights from techniques such as ligand-assisted protein structure (LAPS) and the discovery of novel allosteric modulators are significant additions for better understanding of the endocannabinoid system. There has also been a plethora of fluorescent conjugates that have been assessed for their binding to cannabinoid receptors as well as their potential for cellular imaging. More recently, bifunctional probes containing either fluorophores or electrophilic tags are becoming more prevalent in the literature. Collectively, these molecular tools are invaluable in demonstrating target engagement within the human endocannabinoid system.

show abstract

“…This probe allows monitoring the endogenous CB 2 expression and engagement in human cells [42] . Moreover, further extension of this work has led to the description of a set of highly specific azide‐bearing fluorescent probes based on the scaffold of the selective CB 2 ligand HU308 (Figure 7) that have been successfully used in a variety of applications and cell types [43] …”

Section: Affinity Probesmentioning

confidence: 99%

Chemistry for the Identification of Therapeutic Targets: Recent Advances and Future Directions

et al. 2021

View full text Add to dashboard Cite

The increasing availability of chemical probes for specific applications in chemical biology and medicinal chemistry is extensively facilitating our knowledge of biological systems. This understanding has fundamental implications in the development of new efficacious therapeutic strategies. Nowadays, successful drug discovery programs consider identification, engagement, and endogenous levels of the intended therapeutic target from the earliest preclinical stages. All these aspects are addressed with the use of chemical probes, compounds that rely on the use of a particular chemical reaction to give an answer to a specific biological question. In this review we will describe the underlying chemistry and the most recent advances experienced in the field together with their most important biomedical applications.

show abstract

Development of High-Specificity Fluorescent Probes to Enable Cannabinoid Type 2 Receptor Studies in Living Cells

Cited by 34 publications

References 70 publications

Lighting Up the Plasma Membrane: Development and Applications of Fluorescent Ligands for Transmembrane Proteins

Lighting Up the Plasma Membrane: Development and Applications of Fluorescent Ligands for Transmembrane Proteins

Imaging Cannabinoid Receptors: A Brief Collection of Covalent and Fluorescent Probes for CB

Chemistry for the Identification of Therapeutic Targets: Recent Advances and Future Directions

Contact Info

Product

Resources

About