Abstract

Chemokines and their G-protein-coupled receptors play a diverse role in immune defence by controlling the migration, activation and survival of immune cells1. They are also involved in viral entry, tumour growth and metastasis and hence are important drug targets in a wide range of diseases2,3. Despite very significant efforts by the pharmaceutical industry to develop drugs, with over 50 small-molecule drugs directed at the family entering clinical development, only two compounds have reached the market: maraviroc (CCR5) for HIV infection and plerixafor (CXCR4) for stem-cell mobilization4. The high failure rate may in part be due to limited understanding of the mechanism of action of chemokine antagonists and an inability to optimize compounds in the absence of structural information5. CC chemokine receptor type 9 (CCR9) activation by CCL25 plays a key role in leukocyte recruitment to the gut and represents a therapeutic target in inflammatory bowel disease6. The selective CCR9 antagonist vercirnon progressed to phase 3 clinical trials in Crohn’s disease but efficacy was limited, with the need for very high doses to block receptor activation6. Here we report the crystal structure of the CCR9 receptor in complex with vercirnon at 2.8 Å resolution. Remarkably, vercirnon binds to the intracellular side of the receptor, exerting allosteric antagonism and preventing G-protein coupling. This binding site explains the need for relatively lipophilic ligands and describes another example of an allosteric site on G-protein-coupled receptors7 that can be targeted for drug design, not only at CCR9, but potentially extending to other chemokine receptors.

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Acknowledgements

We thank D. Axford, R. Owen and D. Sherrell at I24, Diamond Light Source, Oxford, UK, for technical support. We thank colleagues at Heptares Therapeutics for suggestions and comments, and G. Brown and S. Bucknell for assistance in radioligand preparation.

Author information

Author notes

    • Christine Oswald
    • , Mathieu Rappas
    •  & James Kean

    These authors contributed equally to this work.

Affiliations

  1. Heptares Therapeutics Ltd, BioPark, Broadwater Road, Welwyn Garden City, Hertfordshire AL7 3AX, UK

    • Christine Oswald
    • , Mathieu Rappas
    • , James Kean
    • , Andrew S. Doré
    • , James C. Errey
    • , Kirstie Bennett
    • , Francesca Deflorian
    • , John A. Christopher
    • , Ali Jazayeri
    • , Jonathan S. Mason
    • , Miles Congreve
    • , Robert M. Cooke
    •  & Fiona H. Marshall

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Contributions

J.K. and A.J. devised and performed the conformational thermostabilization and mutagenesis of the receptor, characterized expression constructs and performed radioligand binding analysis of mutants. Computational analysis of the structure and modelling was performed by F.D. and J.S.M. A.S.D. established the platform/protocols for LCP crystallization and solved the structure. J.C.E. supported expression and scouted purification of the final StaR. M.R. designed and characterized all constructs, collected and processed X-ray diffraction data and solved the structure. C.O. optimized purification, performed LCP crystallization, harvested crystals, collected and processed X-ray diffraction data, and solved and refined the structure. K.B. performed and analysed the pharmacology data. J.A.C. and M.C. identified and sourced the chemical compound(s) used in the study. Project management was performed by J.A.C., R.M.C. and F.H.M. The manuscript was prepared by A.S.D., C.O., F.D., M.C. and F.H.M. All authors contributed to the final editing and approval of the manuscript.

Competing interests

The authors are employees of Heptares Therapeutics and are shareholders of Sosei Group, the parent company of Heptares. Heptares is a drug discovery and development company working in the field of G-protein-coupled receptor structure-based drug design.

Corresponding author

Correspondence to Fiona H. Marshall.

Extended data

Supplementary information

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  1. 1.

    Supplementary Figure

    This file contains Supplementary Figure 1, preparation of [3H]vercirnon and vercirnon.

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DOI

https://doi.org/10.1038/nature20606

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