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A designer molecular chaperone against transmissible spongiform encephalopathy slows disease progression in mice and macaques

Nature Biomedical Engineering volume 3, pages 206–219 (2019)Cite this article

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Abstract

Transmissible spongiform encephalopathies (TSEs) are fatal neurodegenerative diseases that lack therapeutic solutions. Here, we show that the molecular chaperone (N,N′-([cyclohexylmethylene]di-4,1-phenylene)bis(2-[1-pyrrolidinyl]acetamide)), designed via docking simulations, molecular dynamics simulations and quantum chemical calculations, slows down the progress of TSEs. In vitro, the designer molecular chaperone stabilizes the normal cellular prion protein, eradicates prions in infected cells, prevents the formation of drug-resistant strains and directly inhibits the interaction between prions and abnormal aggregates, as shown via real-time quaking-induced conversion and in vitro conversion NMR. Weekly intraperitoneal injection of the chaperone in prion-infected mice prolonged their survival, and weekly intravenous administration of the compound in macaques infected with bovine TSE slowed down the development of neurological and psychological symptoms and reduced the concentration of disease-associated biomarkers in the animals’ cerebrospinal fluid. The de novo rational design of chaperone compounds could lead to therapeutics that can bind to different prion protein strains to ameliorate the pathology of TSEs.

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Fig. 1: Logical design of MC and its anti-prion activity.
Fig. 2: Inhibition of the RT-QuIC process.
Fig. 3: IVC-NMR and the detection of TICs.
Fig. 4: Anti-prion effects of MC.
Fig. 5: Treatment of prion-infected mice with MC.
Fig. 6: Treatment of BSE-infected macaques with MC.
Fig. 7: Digital pathological analyses of BSE-infected macaques with or without MC treatment.

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Code availability

The PAICS codes are available at www.paics.net/index_e.html.

Data availability

The authors declare that all data supporting the findings of this study are available within the paper and its Supplementary Information.

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Acknowledgements

We thank M. Fukushima at the Translational Research Informatics Center and H. Mizusawa at the National Center of Neurology and Psychiatry for fruitful discussion. We also thank R. Honda, T. Saeki, M. Horii and S. Hori for providing technical help. K.K. was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and by grants from the Ministry of Health, Labour and Welfare. The study was also supported by a grant from the Practical Research Project for Rare/Intractable Diseases of the Japan Agency for Medical Research and Development.

Author information

Author notes

  1. Keiichi Yamaguchi

    Present address: Institute for Protein Research, Osaka University, Osaka, Japan

  2. Fumiko Ono

    Present address: Faculty of Veterinary Medicine, Okayama University of Science, Imabari, Japan

  3. Hiroaki Shibata

    Present address: Center for Development of Advanced Medical Technology, Jichi Medical University, Shimotsuke, Japan

  4. Takeshi Ishikawa

    Present address: Graduate School of Science and Engineering, Kagoshima University, Kagoshima, Japan

  5. These authors contributed equally: Keiichi Yamaguchi, Yuji O. Kamatari, Fumiko Ono, Takayuki Fuse.

Authors and Affiliations

  1. Center for Emerging Infectious Diseases, Gifu University, Gifu, Japan

    Keiichi Yamaguchi, Yuji O. Kamatari, Abdelazim Elsayed Elhelaly, Mayuko Fukuoka, Tsutomu Kimura, Junji Hosokawa-Muto, Takeshi Ishikawa & Kazuo Kuwata

  2. United Graduate School of Drug Discovery and Medical Information Sciences, Gifu University, Gifu, Japan

    Keiichi Yamaguchi, Abdelazim Elsayed Elhelaly, Mayuko Fukuoka & Kazuo Kuwata

  3. Life Science Research Center, Gifu University, Gifu, Japan

    Yuji O. Kamatari

  4. Faculty of Animal Crisis Management, Chiba Institute of Science, Choshi, Japan

    Fumiko Ono

  5. Tsukuba Primate Research Center, National Institutes of Biomedical Innovation, Health and Nutrition, Tsukuba, Japan

    Hiroaki Shibata

  6. Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan

    Takayuki Fuse, Takeshi Ishikawa, Daisuke Ishibashi & Noriyuki Nishida

  7. Department of Food Hygiene and Control, Faculty of Veterinary Medicine, Suez Canal University, Ismalia, Egypt

    Abdelazim Elsayed Elhelaly

  8. Department of Chemistry, Faulty of Science Division II, Tokyo University of Science, Tokyo, Japan

    Tsutomu Kimura

  9. First Department of Forsenic Science, National Research Institute of Police Science, Kashiwa, Japan

    Junji Hosokawa-Muto

  10. Department of Pathology, National Institute of Infectious Disease, Tokyo, Japan

    Minoru Tobiume

  11. Department of Biostatistics, School of Public Health, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan

    Yoshinori Takeuchi & Yutaka Matsuyama

  12. Department of Gene and Development, Graduate School of Medicine, Gifu University, Gifu, Japan

    Kazuo Kuwata

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  1. Keiichi Yamaguchi
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Contributions

K.Y. prepared the 15N-labelled and non-labelled recombinant prions, and performed the IVC-NMR measurements and analysis. Y.O.K. measured and analysed the binding data using NMR and SPR. F.O. and H.S. conducted the in vivo treatment study. D.I. and M.T. conducted the pathological examinations for mice and macaques, respectively. T.F. and N.N. analysed Tau proteins in the central nervous system. T.K. established the method for the synthesis of the anti-prion compounds. J.H-M., A.E.E. and M.F. performed the in vitro, ex vivo and in vivo experiments. T.I. coded the FMO programme PAICS using the programming language C. Y.T. and Y.M. performed the statistical analysis. K.K. supervised the project, analysed the data and wrote the manuscript.

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Correspondence to Kazuo Kuwata.

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Reporting summary

Supplementary Video 1

Prion infected macaque (number 6, control) at 19.7 m.p.i.

Supplementary Video 2

Prion infected macaque (number 3, BOS) at 19.7 m.p.i.

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Yamaguchi, K., Kamatari, Y.O., Ono, F. et al. A designer molecular chaperone against transmissible spongiform encephalopathy slows disease progression in mice and macaques. Nat Biomed Eng 3, 206–219 (2019). https://doi.org/10.1038/s41551-019-0349-8

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