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Inhibiting a promiscuous GPCR: iterative discovery of bitter taste receptor ligands

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Abstract

The human GPCR family comprises circa 800 members, activated by hundreds of thousands of compounds. Bitter taste receptors, TAS2Rs, constitute a large and distinct subfamily, expressed orally and extra-orally and involved in physiological and pathological conditions. TAS2R14 is the most promiscuous member, with over 150 agonists and 3 antagonists known prior to this study. Due to the scarcity of inhibitors and to the importance of chemical probes for exploring TAS2R14 functions, we aimed to discover new ligands for this receptor, with emphasis on antagonists. To cope with the lack of experimental structure of the receptor, we used a mixed experimental/computational methodology which iteratively improved the performance of the predicted structure. The increasing number of active compounds, obtained here through experimental screening of FDA-approved drug library, and through chemically synthesized flufenamic acid derivatives, enabled the refinement of the binding pocket, which in turn improved the structure-based virtual screening reliability. This mixed approach led to the identification of 10 new antagonists and 200 new agonists of TAS2R14, illustrating the untapped potential of rigorous medicinal chemistry for TAS2Rs. 9% of the ~ 1800 pharmaceutical drugs here tested activate TAS2R14, nine of them at sub-micromolar concentrations. The iterative framework suggested residues involved in the activation process, is suitable for expanding bitter and bitter-masking chemical space, and is applicable to other promiscuous GPCRs lacking experimental structures.

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Data availability

The updated TAS2R14 ligands dataset and the TAS2R14 models generated within this study are included within the Supplementary Information and will be incorporated within the next release of BitterDB https://bitterdb.agri.huji.ac.il/dbbitter.php.

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Acknowledgements

MYN, LP, EM, and FF are members of COST action ERNEST (CA18133).

Funding

This research was partially supported by the German Research Foundation Grants Gm 13/12 (to PG and MYN), and ISF 494/16 (to MYN). Lady Davis to FF and Excellence Fellowship from the Hebrew University Center for Nanoscience and Nanotechnology to FF, EM, and LP are gratefully acknowledged.

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Authors and Affiliations

  1. The Institute of Biochemistry, Food Science and Nutrition, Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel

    Fabrizio Fierro, Lior Peri, Tamir Dingjan, Eitan Margulis & Masha Y Niv

  2. Department of Chemistry and Pharmacy, Medicinal Chemistry, Friedrich-Alexander-Universität Erlangen-Nürnberg, Nikolaus-Fiebiger-Str. 10, 91058, Erlangen, Germany

    Harald Hübner, Alina Tabor-Schkade, Lukas Waterloo, Stefan Löber, Tara Pfeiffer, Dorothee Weikert & Peter Gmeiner

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  1. Fabrizio Fierro
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  2. Lior Peri
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  4. Alina Tabor-Schkade
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  10. Eitan Margulis
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  11. Peter Gmeiner
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  12. Masha Y Niv
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Contributions

Study conception and design: MYN and PG. Computational methodologies: FF, TD, EM. Experimental test for compounds suggested by the computational methodology: LP. Chemical synthesis of flufenamic acid derivatives: LW. Activation essays of the flufenamic acid derivatives: HH, DW. Screening of the FDA approved drugs library: ATS, DW. Analysis and interpretation of results: FF, LP, MYN, HH. Draft manuscript preparation: FF. Draft manuscript review and editing: FF, LP, TP, SL, MYN, PG.

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Correspondence to Peter Gmeiner or Masha Y Niv.

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Fierro, F., Peri, L., Hübner, H. et al. Inhibiting a promiscuous GPCR: iterative discovery of bitter taste receptor ligands. Cell. Mol. Life Sci. 80, 114 (2023). https://doi.org/10.1007/s00018-023-04765-0

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