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Deschloroclozapine, a potent and selective chemogenetic actuator enables rapid neuronal and behavioral modulations in mice and monkeys

Nature Neuroscience volume 23, pages 1157–1167 (2020)Cite this article

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Abstract

The chemogenetic technology designer receptors exclusively activated by designer drugs (DREADDs) afford remotely reversible control of cellular signaling, neuronal activity and behavior. Although the combination of muscarinic-based DREADDs with clozapine-N-oxide (CNO) has been widely used, sluggish kinetics, metabolic liabilities and potential off-target effects of CNO represent areas for improvement. Here, we provide a new high-affinity and selective agonist deschloroclozapine (DCZ) for muscarinic-based DREADDs. Positron emission tomography revealed that DCZ selectively bound to and occupied DREADDs in both mice and monkeys. Systemic delivery of low doses of DCZ (1 or 3 μg per kg) enhanced neuronal activity via hM3Dq within minutes in mice and monkeys. Intramuscular injections of DCZ (100 μg per kg) reversibly induced spatial working memory deficits in monkeys expressing hM4Di in the prefrontal cortex. DCZ represents a potent, selective, metabolically stable and fast-acting DREADD agonist with utility in both mice and nonhuman primates for a variety of applications.

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Fig. 1: DCZ selectively binds to DREADDs.
Fig. 2: [11C]DCZ PET visualizes DREADD expression and measures the agonist dose–occupancy relationship.
Fig. 3: Time–concentration profiles of DCZ and its metabolites in monkeys and mice.
Fig. 4: In vitro potency and in vivo efficacy of DCZ for DREADDs.
Fig. 5: DCZ rapidly drives activation of hM3Dq-expressing neuronal population in monkey.
Fig. 6: DCZ selectively induces metabolic changes in an hM3Dq-expressing region in a dose-dependent manner.
Fig. 7: DCZ selectively and rapidly induces spatial working-memory deficit in monkeys expressing hM4Di in the PFC.

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Data availability

The data that support the findings of this study are available from the corresponding authors upon reasonable request.

Code availability

The code to generate the results and the figures of this study are available from the corresponding authors upon reasonable request.

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Acknowledgements

We thank R. Suma, J. Kamei, R. Yamaguchi, Y. Matsuda, Y. Sugii, A. Maruyama, T. Okauchi, T. Kokufuta, Y. Iwasawa, T. Watanabe, A. Tanizawa, S. Shibata, N. Nitta, Y. Ozawa, M. Fujiwara, M. Nakano, T. Minamihisamatsu, S. Uchida and S. Sasaki for their technical assistance. We also thank S. Hiura for 3D printing of the grids. This study was supported by the following grants and organizations: MEXT/JSPS KAKENHI grant numbers JP15H05917, JP15K12772 and JP18H04037 (to T.M.), JP16H02454 (to M. Takada), JP19K08138 (to Y.N.), and JP18H05018, JP19K07811 and JP20H04596 (to N.M.); AMED grant numbers JP20dm0107146 (to T.M.), JP19dm0207003 (to M. Takada), JP20dm0107094 and JP18dm0207007 (to T.S.), JP20dm0307021 (to K.-i.I.), JP20dm0307007 (to T.H.), and JP20dm0207072 (to M.H.); JST PRESTO grant number JPMJPR1683 (to K.-i.I.); QST President’s Strategic Grant (Creative Research) (to N.M.); the Cooperative Research Program at PRI, Kyoto University; the National Bio-Resource Project ‘Japanese Monkeys’ of MEXT, Japan; and U24DK116195, the NIMH Psychoactive Drug Screening Program and the Michael Hooker Distinguished Professorship to B.L.R.

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Author notes

  1. These authors contributed equally: Yuji Nagai and Naohisa Miyakawa.

Authors and Affiliations

  1. Department of Functional Brain Imaging, National Institute of Radiological Sciences, National Institutes for Quantum and Radiological Science and Technology, Chiba, Japan

    Yuji Nagai, Naohisa Miyakawa, Hiroyuki Takuwa, Yukiko Hori, Kei Oyama, Bin Ji, Manami Takahashi, Takuya Urushihata, Toshiyuki Hirabayashi, Atsushi Fujimoto, Koki Mimura, Chie Seki, Maiko Ono, Masafumi Shimojo, Tetsuya Suhara, Makoto Higuchi & Takafumi Minamimoto

  2. Department of Pharmacology, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC, USA

    Xi-Ping Huang, Samuel T. Slocum, Jeffrey F. DiBerto, Justin G. English & Bryan L. Roth

  3. Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    Xi-Ping Huang & Bryan L. Roth

  4. Mount Sinai Center for Therapeutics Discovery, Departments of Pharmacological Sciences and Oncological Sciences, Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA

    Yan Xiong, Jing Liu & Jian Jin

  5. Systems Neuroscience Section, Primate Research Institute, Kyoto University, Inuyama, Japan

    Ken-ichi Inoue & Masahiko Takada

  6. PRESTO, Japan Science and Technology Agency, Kawaguchi, Saitama, Japan

    Ken-ichi Inoue

  7. Department of Radiopharmaceuticals Development, National Institute of Radiological Sciences, National Institutes for Quantum and Radiological Science and Technology, Chiba, Japan

    Katsushi Kumata & Ming-Rong Zhang

  8. Department of Neurology, Keio University School of Medicine, Tokyo, Japan

    Yutaka Tomita & Jin Nakahara

  9. National Institute of Mental Health Psychoactive Drug Screening Program (NIMH PDSP), Department of Pharmacology, University of North Carolina at Chapel Hill Medical School, Chapel Hill, NC, USA

    Bryan L. Roth

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Contributions

Conceptualization: T.M. Formal analysis: Y.N., N.M., K.O., K.M. and T.M. Investigation: Y.N., N.M., H.T., Y.H., K.O., B.J., M. Takahashi, X.-P.H., S.T.S., J.F.D., T.U., A.F., J.G.E., K.K., C.S., M.O. and M.S. Resources: B.J., Y.X., J.L., K.-i.I., Y.T., J.N., M. Takada and J.J. Writing (original draft): Y.N., N.M. and T.M. Visualization: Y.N., N.M., H.T., Y.H., K.O., B.J., K.M. and T.M. Supervision: M.-R.Z., T.S., M. Takada, M.H., J.J., B.L.R. and T.M. Project administration: B.L.R. and T.M. Funding acquisition: Y.N., N.M., T.H., K.I., M. Takada, T.S., M.H., J.J., B.L.R., and T.M. Writing (review and editing): all authors.

Corresponding authors

Correspondence to Bryan L. Roth or Takafumi Minamimoto.

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Competing interests

Y.N., N.M., B.J., T.S., M.H. and T.M. are named as inventors on a patent application (PCT/JP2019/024834; status: patent pending) claiming subject matter related to the results described in this paper. The remaining authors declare no competing interests.

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Peer review information Nature Neuroscience thanks Mikhail Shapiro and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Nagai, Y., Miyakawa, N., Takuwa, H. et al. Deschloroclozapine, a potent and selective chemogenetic actuator enables rapid neuronal and behavioral modulations in mice and monkeys. Nat Neurosci 23, 1157–1167 (2020). https://doi.org/10.1038/s41593-020-0661-3

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