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Inhibition of the Vesicular Glutamate Transporter (VGLUT) with Congo Red Analogs: New Binding Insights

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Abstract

The vesicular glutamate transporter (VGLUT) facilitates the uptake of glutamate (Glu) into neuronal vesicles. VGLUT has not yet been fully characterized pharmacologically but a body of work established that certain azo-dyes bearing two Glu isosteres via a linker were potent inhibitors. However, the distance between the isostere groups that convey potent inhibition has not been delineated. This report describes the synthesis and pharmacologic assessment of Congo Red analogs that contain one or two glutamate isostere or mimic groups; the latter varied in the interatomic distance and spacer properties to probe strategic binding interactions within VGLUT. The more potent inhibitors had two glutamate isosteres symmetrically linked to a central aromatic group and showed IC50 values ~ 0.3–2.0 μM at VGLUT. These compounds contained phenyl, diphenyl ether (PhOPh) or 1,2-diphenylethane as the linker connecting 4-aminonaphthalene sulfonic acid groups. A homology model for VGLUT2 using d-galactonate transporter (DgoT) to dock and identify R88, H199 and F219 as key protein interactions with Trypan Blue, Congo Red and selected potent analogs prepared and tested in this report.

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Abbreviations

Glu:

Glutamate

VGLUT:

Vesicular glutamate transporter

DgoT:

d-galactonate transporter

TB:

Trypan Blue

CR:

Congo Red

4-ANS:

4-Aminonaphthalene sulfonic acid

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Acknowledgements

The authors wish to thank Dr David Holley for assistance with the molecular modeling and the University of Montana’s Center for Structural & Functional Neuroscience. Research in the authors laboratories is funded by a grant to The Center for Biomolecular and Structural Dynamics from the National Institutes of Health, P20GM103546.

Funding

Financial support for University of Montana Molecular Computational and Magnetic Resonance Core Facilities utilized in this study was provided by NIH NIGMS P20GM103546 (S. Sprang, PI) and gratefully acknowledged.

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Author notes

  1. David M. Hitt

    Present address: Department of Chemistry, Carroll College, 1601 N Benton Ave., Helena, MT, 59625, USA

  2. Jeffery D. Zwicker

    Present address: Deciphera Pharmaceuticals, 643 Massachusetts St, Lawrence, KS, 66044, USA

Authors and Affiliations

  1. Department of Biomedical and Pharmaceutical Sciences, Center for Structural and Functional Neuroscience, College of Health Professions and Biomedical Sciences, University of Montana, Missoula, MT, 59812, USA

    David M. Hitt, Jeffery D. Zwicker, Chih-Kai Chao, Sarjubhai A. Patel, John M. Gerdes, Richard J. Bridges & Charles M. Thompson

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Contributions

The manuscript was written by C.M.T. with contributions by all authors. DH, JZ, SP and C-KC conducted the investigation, methodology and formal analyses. Conceptualization was conducted by JG, RB and CT. All authors have given approval to the final version of the manuscript.

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Correspondence to Charles M. Thompson.

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Hitt, D.M., Zwicker, J.D., Chao, CK. et al. Inhibition of the Vesicular Glutamate Transporter (VGLUT) with Congo Red Analogs: New Binding Insights. Neurochem Res 46, 494–503 (2021). https://doi.org/10.1007/s11064-020-03182-0

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