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Small-molecule factor D inhibitors targeting the alternative complement pathway

Nature Chemical Biology volume 12, pages 1105–1110 (2016)Cite this article

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Abstract

Complement is a key component of the innate immune system, recognizing pathogens and promoting their elimination. Complement component 3 (C3) is the central component of the system. Activation of C3 can be initiated by three distinct routes—the classical, the lectin and the alternative pathways—with the alternative pathway also acting as an amplification loop for the other two pathways. The protease factor D (FD) is essential for this amplification process, which, when dysregulated, predisposes individuals to diverse disorders including age-related macular degeneration and paroxysmal nocturnal hemoglobinuria (PNH). Here we describe the identification of potent and selective small-molecule inhibitors of FD. These inhibitors efficiently block alternative pathway (AP) activation and prevent both C3 deposition onto, and lysis of, PNH erythrocytes. Their oral administration inhibited lipopolysaccharide-induced AP activation in FD-humanized mice. These data demonstrate the feasibility of inhibiting the AP with small-molecule antagonists and support the development of FD inhibitors for the treatment of complement-mediated diseases.

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Figure 1: Structure-based evolution of reversible small-molecule human FD inhibitors.
Figure 2: Identification and characterization of potent and selective human FD inhibitor 6.
Figure 3: Generation of human factor D knock-in mice.
Figure 4: Inhibitor (6) blocks LPS-induced systemic and ocular AP activation in C57Bl/6 mice expressing human FD.
Figure 5: Inhibitors of FD prevent C3 deposition on and hemolysis of PNH erythrocytes.

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Acknowledgements

We thank T. Zoller for the preparation of the proline-based compound library, J. Wirsching, T. Doll and A. Isken for their assistance in generating the human FD knock-in mice, N. Buchanan for coordinating the maintenance of the mouse colony, F. Zink for assistance in preparing FD and KLK7 inhibitor complex crystals, S. Kapps for preparation of 15N-labeled human FD, C. Towler for small molecule crystallization experiments, C. Dentel and F. Tritsch for chemical syntheses, U. Argikar, A. Brown, S. Bailey and G. Marsh for pharmacokinetic and bioanalytical support, A. De Erkenez for performing human whole blood assays, and L. Ferrara for performing the AP hemolytic assay.

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Authors and Affiliations

  1. Novartis Institutes for BioMedical Research, Novartis Pharma AG, Novartis Campus, Basel, Switzerland

    Jürgen Maibaum, Anna Vulpetti, Nils Ostermann, Simon Rüdisser, Edwige Lorthiois, Paul Erbel, Bernd Kinzel, Samuel Barbieri, Julia Wagner, Corinne Durand, Kamal Fettis, Solene Dussauge, Nicola Hughes, Ulrich Hommel, Bernd Gerhartz, Frederic Cumin, Stefanie Flohr, Anna Schubart & Jörg Eder

  2. Novartis Institutes for BioMedical Research, Cambridge, Massachusetts, USA

    Sha-Mei Liao, Omar Delgado, Ty Gould, Bruce Jaffee & Karen Anderson

  3. Evonik Japan Co., Shinjuku-ku, Tokyo, Japan

    Stefan Randl

  4. Pharma Research and Early Development, Roche Innovation Center Basel, Basel, Switzerland

    Fabrice A Kolb

  5. Drug Discovery Department, Actelion Pharmaceuticals Ltd., Allschwil, Switzerland

    Aengus Mac Sweeney

  6. Institute of Infection and Immunity, School of Medicine, Cardiff University, Henry Wellcome Building, Heath Park, Cardiff, UK

    Richard Harrison

  7. Department of Clinical Medicine and Surgery, Division of Hematology, University of Naples, Naples, Italy

    Antonio Maria Risitano

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  1. Jürgen Maibaum
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Contributions

N.O. and A.M.S. solved the structures by X-ray crystallography. S. Randl, U.H., A.V., E.L. and J.M. designed the FD structure-based inhibitors. S.D., C.D. and K.F. synthesized compounds. S. Rüdisser and P.E. performed NMR experiments for screening and binding affinity measurements. N.H. and P.E. conducted SPR experiments. F.C. generated the biochemical assay data. A.S. and J.W. performed the PNH surrogate assay and analyzed the data. A.M.R. performed the assays using erthrocytes from PNH patient and analyzed the data. S.B., B.K. and F.A.K. generated the human FD knock-in mice. B.G. and J.E. produced recombinant proteins. T.G. designed and interpreted pharmacokinetic studies. O.D. and S.-M.L. performed the in vivo studies in mice. S.F. and B.J. conceived experiments and supervised part of the work. J.M., J.E., A.S., R.H. and K.A. conceived experiments, supervised the work, and wrote the manuscript.

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Correspondence to Jürgen Maibaum or Karen Anderson.

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Competing interests

All authors, with the exception of A.M.R., are former or current employees of Novartis and may hold Novartis stock. Experiments reported herein conducted by A.M.R. were funded by Novartis.

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Supplementary Results, Supplementary Tables 1–4 and Supplementary Figures 1–6. (PDF 1383 kb)

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Synthetic Procedures. (PDF 733 kb)

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Maibaum, J., Liao, SM., Vulpetti, A. et al. Small-molecule factor D inhibitors targeting the alternative complement pathway. Nat Chem Biol 12, 1105–1110 (2016). https://doi.org/10.1038/nchembio.2208

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