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TOPKi-NBD: a fluorescent small molecule for tumor imaging

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European Journal of Nuclear Medicine and Molecular Imaging Aims and scope Submit manuscript

Abstract

Purpose

OTS514 is a highly specific inhibitor targeting lymphokine-activated killer T cell-originated protein kinase (TOPK). A fluorescently labeled TOPK inhibitor could be used for tumor delineation or intraoperative imaging, potentially improving patient care.

Methods

Fluorescently labeled OTS514 was obtained by conjugating the fluorescent small molecule NBD to the TOPK inhibitor. HCT116 colorectal cancer cells were used to generate tumors in NSG mice for in vivo studies. Images were generated in vitro using confocal microscopy and ex vivo using an IVIS Spectrum.

Results

OTS514 was successfully conjugated to a fluorescent sensor and validated in vitro, in vivo, and ex vivo. The labeling reaction led to TOPKi-NBD with 67% yield and 97% purity after purification. We were able to test binding properties of TOPKi-NBD to its target, TOPK, and compared them to the precursor inhibitor. EC50s showed similar target affinities for TOPKi-NBD and the unlabeled OTS514. TOPKi-NBD showed specific tumor uptake after systemic administration and was microscopically detectable inside cancer cells ex vivo. Blocking controls performed with an excess of the unlabeled OTS514 confirmed specificity of the compound. Overall, the results represent a first step toward the development of a class of TOPK-specific fluorescent inhibitors for in vivo imaging and tumor delineation.

Conclusions

TOPK has the potential to be a new molecular target for cancer-specific imaging in a large variety of tumors. This could lead to broad applications in vitro and in vivo.

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Acknowledgments

The authors wish to acknowledge the support of Memorial Sloan Kettering Cancer Center’s Small Animal Imaging Core Facility, Radiochemistry & Molecular Imaging Probes Core Facility, Integrated Genomics Core Facility, Nuclear Magnetic Resonance Core Facility, Media Preparation Core Facility, and Molecular Cytology Core Facility. We wish to thank Dr. Pat Zanzonico and Ms. Valerie Longo for technical support with IVIS imaging.

Funding

This work was supported by National Institutes of Health grants R01 CA204441 (T.R.), P30 CA008748, and the Memorial Sloan Kettering Imaging and Radiation Sciences Program (T.R.).

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Authors and Affiliations

  1. Department of Radiology, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, NY, 10065, USA

    Giacomo Pirovano, Sheryl Roberts & Thomas Reiner

  2. Department of Radiology, Weill Cornell Medical College, 1300 York Avenue, New York, NY, 10065, USA

    Thomas Reiner

  3. Chemical Biology Program, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, NY, 10065, USA

    Thomas Reiner

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  1. Giacomo Pirovano
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Correspondence to Thomas Reiner.

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Conflict of interest

T.R. is shareholder of Summit Biomedical Imaging, LLC and paid consultant for Theragnostics, Inc. G.P. and S.R. declare no conflict of interest.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Pirovano, G., Roberts, S. & Reiner, T. TOPKi-NBD: a fluorescent small molecule for tumor imaging. Eur J Nucl Med Mol Imaging 47, 1003–1010 (2020). https://doi.org/10.1007/s00259-019-04608-w

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  • DOI: https://doi.org/10.1007/s00259-019-04608-w

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