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(-)-Stepholidine is a potent pan-dopamine receptor antagonist of both G protein- and β-arrestin-mediated signaling

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Abstract

Rationale

(−)-Stepholidine is a tetrahydroberberine alkaloid that is known to interact with dopamine receptors and has also been proposed as a novel antipsychotic agent. Its suggested novelty lies in the fact that it has been proposed to have D1-like receptor agonist and D2-like receptor antagonist properties. Thus, it might be effective in treating both positive and negative (cognition) symptoms of schizophrenia. However, its activity on specific dopamine receptor subtypes has not been clarified, especially with respect to its ability to activate D1-like receptors.

Objectives

We wished to examine the affinity and functional activity of (−)-stepholidine at each of the human dopamine receptor subtypes expressed in a defined cellular environment.

Methods

D1–D5 dopamine receptors were stably expressed in cell lines and their interactions with (−)-stepholidine were examined using radioligand binding and various functional signaling assays. Radioligand binding assays were also performed using bovine striatal membranes.

Results

(−)-Stepholidine exhibited high (nM) affinity for D1 and D5 receptors, somewhat lower (two- to four-fold) affinity for D2 and D3 receptors, and low micromolar affinity for D4 receptors. Functionally, (−)-stepholidine was ineffective in activating G protein-mediated signaling of D1-like and D2 receptors and was also ineffective in stimulating β-arrestin recruitment to any dopamine receptor subtype. It did, however, antagonize all of these responses. It also antagonized D1–D2 heteromer-mediated Ca2+ mobilization. Radioligand binding assays of D1-like receptors in brain membranes also indicated that (−)-stepholidine binds to the D1 receptor with antagonist-like properties.

Conclusions

(−)-Stepholidine is a pan-dopamine receptor antagonist and its in vivo effects are largely mediated through dopamine receptor blockade with potential cross-talk to other receptors or signaling proteins.

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Acknowledgments

This work was supported, in part, by the Intramural Research Program of the National Institute of Neurological Disorders and Stroke (NINDS). All authors declare no conflicts of interest. The authors would like to thank Dr. Peter W. Dematteo (NIDA) for optical rotation analysis and NMR confirmation of (−)stepholidine batches, and Mr. Bryce Adams for technical assistance.

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Authors and Affiliations

  1. Molecular Neuropharmacology Section, National Institute of Neurological Disorders and Stroke, National Institutes of Health, 5625 Fishers Lane, Room 4S-04, Bethesda, MD, 20892-9405, USA

    Julie A. Meade, R. Benjamin Free, Nicole R. Miller, Lani S. Chun, Trevor B. Doyle, Amy E. Moritz, Jennie L. Conroy & David R. Sibley

  2. Cell, Molecular, Developmental Biology & Biophysics Program, Johns Hopkins University, Baltimore, MD, USA

    Lani S. Chun

  3. Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, West Lafayette, IN, USA

    Trevor B. Doyle & Val J. Watts

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  1. Julie A. Meade
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  2. R. Benjamin Free
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  3. Nicole R. Miller
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  4. Lani S. Chun
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  5. Trevor B. Doyle
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  9. David R. Sibley
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Correspondence to David R. Sibley.

Additional information

Julie A. Meade and R. Benjamin Free contributed equally to this work.

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Meade, J.A., Free, R.B., Miller, N.R. et al. (-)-Stepholidine is a potent pan-dopamine receptor antagonist of both G protein- and β-arrestin-mediated signaling. Psychopharmacology 232, 917–930 (2015). https://doi.org/10.1007/s00213-014-3726-8

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