A Highly Selective and Versatile Probe Platform for Visualization of Monoacylglycerol Lipase

Angewandte Chemie International Edition, vol. 64, pp. e202413405

Abstract

Monoacylglycerol lipase (MAGL) is a key enzyme for signal termination in the endocannabinoid system (ECS). MAGL inhibition results in indirect activation of the cannabinoid receptors, which offers unique advantages for the treatment of, e.g., multiple sclerosis, epilepsy, and other neurological disorders. Molecular imaging techniques are valuable tools to overcome the current poor understanding of MAGL's distribution and role in patho- and physiological processes within ECS signaling. Herein, we report the design, synthesis, and validation of highly selective versatile fluorescent and click-chemistry probes for MAGL. Structure-based design combined with a reverse-design approach allowed the development of a structural unit that selectively and effectively recognizes MAGL while offering a versatile platform to attach different fluorophores and further reporter units. In this way, labeled probes with sub-nanomolar potency carrying diverse fluorescent dyes were obtained. Probe affinity and selectivity remained invariant to changes in the fluorophore subunit, showing the remarkable robustness of this platform in delivering tailor-made probes. Highly consistent inhibition across species supports pharmacological model translatability. Extensive profiling and validation in various cellular systems shows the ability of these highly potent and selective probes to elucidate the complex role of MAGL in ECS cellular signaling, inflammatory processes, and disease progression.

Authors 19

1. Axel Hentsch

   Leibniz-Forschungsinstitut für Molekulare Pharmakologie

   Affiliation as printed

   Leibniz Forschungsinstitut für Molekulare Pharmakologie (FMP) Robert-Roessle-Strasse 10 13125 Berlin GER

   These authors contributed equally to this work

2. Mónica Guberman

   Leibniz-Forschungsinstitut für Molekulare Pharmakologie

   Affiliation as printed

   Leibniz Forschungsinstitut für Molekulare Pharmakologie (FMP) Robert-Roessle-Strasse 10 13125 Berlin GER

   These authors contributed equally to this work

3. Silke Radetzki

   Leibniz-Forschungsinstitut für Molekulare Pharmakologie

   Affiliation as printed

   Leibniz Forschungsinstitut für Molekulare Pharmakologie (FMP) Robert-Roessle-Strasse 10 13125 Berlin GER

4. Mirjam C. W. Huizenga Aachen Leiden Institute of Chemistry

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry (LIC) Universiteit Leiden Einsteinweg 55 2333 CC Leiden NL

   Leiden Institute of Chemistry (LIC), Universiteit Leiden, Einsteinweg 55, 2333 CC Leiden, NL

5. Jérôme Paul

   Leibniz-Forschungsinstitut für Molekulare Pharmakologie

   Affiliation as printed

   Leibniz Forschungsinstitut für Molekulare Pharmakologie (FMP) Robert-Roessle-Strasse 10 13125 Berlin GER

6. Jörg Benz

   Roche (Switzerland)

   Affiliation as printed

   Pharma Research & Early Development (pRED) F. Hoffmann-La Roche Ltd Grenzacherstrasse 124 CH-4070 Basel CH

   Pharma Research & Early Development (pRED), F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, CH-4070 Basel, CH

7. Bernd Kuhn

   Roche (Switzerland)

   Affiliation as printed

   Pharma Research & Early Development (pRED) F. Hoffmann-La Roche Ltd Grenzacherstrasse 124 CH-4070 Basel CH

   Pharma Research & Early Development (pRED), F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, CH-4070 Basel, CH

8. Dominik Heer

   Roche (Switzerland)

   Affiliation as printed

   Pharma Research & Early Development (pRED) F. Hoffmann-La Roche Ltd Grenzacherstrasse 124 CH-4070 Basel CH

   Pharma Research & Early Development (pRED), F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, CH-4070 Basel, CH

9. Andreas Topp

   Roche (Switzerland)

   Affiliation as printed

   Pharma Research & Early Development (pRED) F. Hoffmann-La Roche Ltd Grenzacherstrasse 124 CH-4070 Basel CH

   Pharma Research & Early Development (pRED), F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, CH-4070 Basel, CH

10. Ludivine Esteves Gloria

    Roche (Switzerland)

    Affiliation as printed

    Pharma Research & Early Development (pRED) F. Hoffmann-La Roche Ltd Grenzacherstrasse 124 CH-4070 Basel CH

    Pharma Research & Early Development (pRED), F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, CH-4070 Basel, CH

11. Remo Hochstrasser

    Roche (Switzerland)

    Affiliation as printed

    Pharma Research & Early Development (pRED) F. Hoffmann-La Roche Ltd Grenzacherstrasse 124 CH-4070 Basel CH

    Pharma Research & Early Development (pRED), F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, CH-4070 Basel, CH

12. Matthias Wittwer

    Roche (Switzerland)

    Affiliation as printed

    Pharma Research & Early Development (pRED) F. Hoffmann-La Roche Ltd Grenzacherstrasse 124 CH-4070 Basel CH

    Pharma Research & Early Development (pRED), F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, CH-4070 Basel, CH

13. Jens Peter von Kries

    Leibniz-Forschungsinstitut für Molekulare Pharmakologie

    Affiliation as printed

    Leibniz Forschungsinstitut für Molekulare Pharmakologie (FMP) Robert-Roessle-Strasse 10 13125 Berlin GER

14. Ludovic Collin

    Roche (Switzerland)

    Affiliation as printed

    Pharma Research & Early Development (pRED) F. Hoffmann-La Roche Ltd Grenzacherstrasse 124 CH-4070 Basel CH

    Pharma Research & Early Development (pRED), F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, CH-4070 Basel, CH

15. Julie Blaising

    Roche (Switzerland)

    Affiliation as printed

    Pharma Research & Early Development (pRED) F. Hoffmann-La Roche Ltd Grenzacherstrasse 124 CH-4070 Basel CH

    Pharma Research & Early Development (pRED), F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, CH-4070 Basel, CH

16. Mario van der Stelt Aachen Leiden Institute of Chemistry

    Leiden University

    Affiliation as printed

    Leiden Institute of Chemistry (LIC) Universiteit Leiden Einsteinweg 55 2333 CC Leiden NL

    Leiden Institute of Chemistry (LIC), Universiteit Leiden, Einsteinweg 55, 2333 CC Leiden, NL

17. Noa Lipstein

    Leibniz-Forschungsinstitut für Molekulare Pharmakologie

    Affiliation as printed

    Leibniz Forschungsinstitut für Molekulare Pharmakologie (FMP) Robert-Roessle-Strasse 10 13125 Berlin GER

18. Uwe Grether corresponding

    Roche (Switzerland)

    Affiliation as printed

    Pharma Research & Early Development (pRED) F. Hoffmann-La Roche Ltd Grenzacherstrasse 124 CH-4070 Basel CH

    Pharma Research & Early Development (pRED), F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, CH-4070 Basel, CH

19. Marc Nazaré corresponding

    Leibniz-Forschungsinstitut für Molekulare Pharmakologie

    Affiliation as printed

    Leibniz Forschungsinstitut für Molekulare Pharmakologie (FMP) Robert-Roessle-Strasse 10 13125 Berlin GER