Photoswitchable Inhibitors to Optically Control Specific Kinase Activity

ACS Chemical Biology, vol. 18, pp. 1378–1387

Abstract

Potent and selective small-molecule inhibitors are valuable tools to elucidate the functions of protein kinases within complex signaling networks. Incorporation of a photoswitchable moiety into the inhibitor scaffold offers the opportunity to steer inhibitor potency with temporal precision, while the challenge of selective inhibition can often be addressed by employing a chemical genetic approach, termed the analog-sensitive method. Here, we combine the perks of these two approaches and report photoswitchable azopyrazoles to target calcium-dependent protein kinase 1 (CDPK1) from Toxoplasma gondii, a kinase naturally susceptible to analog-sensitive kinase inhibitors due to its glycine gatekeeper residue. The most promising azopyrazoles display favorable photochemical properties, thermal stability, and a substantial difference in IC 50 values between both photostationary states. Consequently, the CDPK1 kinase reaction can be controlled dynamically and reversibly by applying light of different wavelengths. Inhibition of CDPK1 by the azopyrazoles drastically relies on the nature of the gatekeeper residue as a successive increase in gatekeeper size causes a concurrent loss of inhibitory activity. Furthermore, two photoswitchable inhibitors exhibit activity against T. gondii and Cryptosporidium parvum infection in a cell culture model, making them a promising addition to the toolbox for dissecting the role of CDPK1 in the infectious cycle with high temporal control. Overall, this work merges the benefits of the analog-sensitive approach and photopharmacology without compromising inhibitory potency and thus holds great promise for application to other protein kinases in the future.

Authors 11

1. Tim Aguirre

   Humboldt-Universität zu Berlin · Leibniz-Forschungsinstitut für Molekulare Pharmakologie

   Affiliation as printed

   Department of Chemistry, Humboldt-Universität zu Berlin, Brook-Taylor-Street 2, 12489 Berlin, Germany

   Leibniz-Forschungsinstitut für Molekulare Pharmakologie, Robert-Rössle-Straße 10, 13125 Berlin, Germany

2. Ellen Teichmann

   Humboldt-Universität zu Berlin

   Affiliation as printed

   Department of Chemistry, Humboldt-Universität zu Berlin, Brook-Taylor-Street 2, 12489 Berlin, Germany

3. Florian Römpp Aachen DWI - Leibniz Institute for Interactive Materials Institute of Technical and Macromolecular Chemistry

   Humboldt-Universität zu Berlin · DWI – Leibniz Institute for Interactive Materials · RWTH Aachen University

   Affiliation as printed

   DWI-Leibniz Institute for Interactive Materials, Forckenbeckstr. 50, 52074 Aachen, Germany

   Department of Chemistry, Humboldt-Universität zu Berlin, Brook-Taylor-Street 2, 12489 Berlin, Germany

   Institute of Technical and Macromolecular Chemistry, RWTH Aachen University, Worringer Weg 2, 52074 Aachen, Germany

4. Ruthey Vivier

   Oregon Health & Science University · Portland VA Medical Center · VA Portland Health Care System

   Affiliation as printed

   School of Medicine Division of Infectious Diseases, Oregon Health & Science University, Portland, 97239 Oregon, United States

   VA Portland Healthcare System, Portland, 97239 Oregon, United States

5. Cole Bryant

   Oregon Health & Science University · Portland VA Medical Center · VA Portland Health Care System

   Affiliation as printed

   School of Medicine Division of Infectious Diseases, Oregon Health & Science University, Portland, 97239 Oregon, United States

   VA Portland Healthcare System, Portland, 97239 Oregon, United States

6. Matthew A. Hulverson

   University of Washington

   Affiliation as printed

   Division of Allergy and Infectious Diseases, Center for Emerging and Re-emerging Infectious Diseases (CERID), Department of Medicine, University of Washington, 750 Republican Street, P.O. Box 358061, Seattle, 98109 Washington, United States

7. Wesley C. Van Voorhis

   University of Washington

   Affiliation as printed

   Division of Allergy and Infectious Diseases, Center for Emerging and Re-emerging Infectious Diseases (CERID), Department of Medicine, University of Washington, 750 Republican Street, P.O. Box 358061, Seattle, 98109 Washington, United States

8. Kayode K. Ojo

   University of Washington

   Affiliation as printed

   Division of Allergy and Infectious Diseases, Center for Emerging and Re-emerging Infectious Diseases (CERID), Department of Medicine, University of Washington, 750 Republican Street, P.O. Box 358061, Seattle, 98109 Washington, United States

9. Joseph Stone Doggett

   Oregon Health & Science University · Portland VA Medical Center · VA Portland Health Care System

   Affiliation as printed

   School of Medicine Division of Infectious Diseases, Oregon Health & Science University, Portland, 97239 Oregon, United States

   VA Portland Healthcare System, Portland, 97239 Oregon, United States

10. Dorothea Fiedler corresponding

    Humboldt-Universität zu Berlin · Leibniz-Forschungsinstitut für Molekulare Pharmakologie

    Affiliation as printed

    Department of Chemistry, Humboldt-Universität zu Berlin, Brook-Taylor-Street 2, 12489 Berlin, Germany

    Leibniz-Forschungsinstitut für Molekulare Pharmakologie, Robert-Rössle-Straße 10, 13125 Berlin, Germany

11. Stefan Hecht corresponding

    Humboldt-Universität zu Berlin

    Affiliation as printed

    Department of Chemistry, Humboldt-Universität zu Berlin, Brook-Taylor-Street 2, 12489 Berlin, Germany