Gram scale preparation of clozapine N-oxide (CNO), a synthetic small molecule actuator for muscarinic acetylcholine DREADDs

Phillip L Van Der Peet, Christian Andre Gunawan, Alaa Abdul Ridha, Sherie Ma, Daniel J Scott, Andrew L Gundlach, Ross Bathgate, Jonathan M. White, Spencer John Williams

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Chemogenetics uses engineered proteins that are controlled by small molecule actuators, allowing in vivo functional studies of proteins with temporal and dose control, and include Designer Receptors Exclusively Activated by Designer Drugs (DREADDs). One major class of DREADDs are mutated muscarinic receptors that are unresponsive to acetylcholine, and are activated by administration of clozapine N-oxide (CNO). However, CNO is available in only small amounts and large scale studies involving animals and multiple cohorts are prohibitively expensive for many investigators. The precursor, clozapine, is also expensive when purchased from specialist suppliers. Here we report: • A simple extraction method of clozapine from commercial tablets;• A simple preparation of CNO from clozapine, and for the first time its single-crystal X-ray structure; and• That the CNO prepared by this method specifically activates the DREADD receptor hM3Dq in vivo.This method provides large quantities of CNO suitable for large-scale DREADD applications that is identical to commercial material.

Original languageEnglish
Pages (from-to)257-267
Number of pages11
JournalMethodsX
Volume5
DOIs
Publication statusPublished - 1 Jan 2018
Externally publishedYes

Keywords

  • Chemical synthesis
  • Chemical-genetics
  • G-protein-coupled receptor (GPCR)

Cite this

Van Der Peet, P. L., Gunawan, C. A., Abdul Ridha, A., Ma, S., Scott, D. J., Gundlach, A. L., ... Williams, S. J. (2018). Gram scale preparation of clozapine N-oxide (CNO), a synthetic small molecule actuator for muscarinic acetylcholine DREADDs. MethodsX, 5, 257-267. https://doi.org/10.1016/j.mex.2018.03.003
Van Der Peet, Phillip L ; Gunawan, Christian Andre ; Abdul Ridha, Alaa ; Ma, Sherie ; Scott, Daniel J ; Gundlach, Andrew L ; Bathgate, Ross ; White, Jonathan M. ; Williams, Spencer John. / Gram scale preparation of clozapine N-oxide (CNO), a synthetic small molecule actuator for muscarinic acetylcholine DREADDs. In: MethodsX. 2018 ; Vol. 5. pp. 257-267.
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Van Der Peet, PL, Gunawan, CA, Abdul Ridha, A, Ma, S, Scott, DJ, Gundlach, AL, Bathgate, R, White, JM & Williams, SJ 2018, 'Gram scale preparation of clozapine N-oxide (CNO), a synthetic small molecule actuator for muscarinic acetylcholine DREADDs' MethodsX, vol. 5, pp. 257-267. https://doi.org/10.1016/j.mex.2018.03.003

Gram scale preparation of clozapine N-oxide (CNO), a synthetic small molecule actuator for muscarinic acetylcholine DREADDs. / Van Der Peet, Phillip L; Gunawan, Christian Andre; Abdul Ridha, Alaa; Ma, Sherie; Scott, Daniel J; Gundlach, Andrew L; Bathgate, Ross; White, Jonathan M.; Williams, Spencer John.

In: MethodsX, Vol. 5, 01.01.2018, p. 257-267.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Van Der Peet, Phillip L

AU - Gunawan, Christian Andre

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AU - Scott, Daniel J

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AU - Bathgate, Ross

AU - White, Jonathan M.

AU - Williams, Spencer John

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