Fructose-based probe molecules for high-throughput screening of GLUT5 transporter inhibitors

Overview

This technology centers on fluorescently labeled fructose mimics that selectively bind to the GLUT5 transporter, enabling real-time monitoring of fructose uptake into cells. Because GLUT5 is overexpressed in certain cancers, these probes support both cancer detection research and the discovery of compounds that inhibit GLUT5 activity. The platform builds on a published assay (ACS Bio & Med Chem Au, 2023) that has been demonstrated using cultured mouse and human breast carcinoma cell lines known to express GLUT5. The current effort focuses on adapting the assay to 96-well plate formats and scalable high-throughput screening (HTS) workflows, with future plans for a dual-color assay that simultaneously monitors fructose and glucose uptake via GLUT5 and GLUT1 respectively.

Technical specifications

Key features:

• Fluorescently labeled fructose mimics with selectivity for the GLUT5 transporter
• Fluorescent readout compatible with standard plate readers for rapid, quantitative measurement of uptake
• Validated assay using EMT6 mouse breast carcinoma and MCT7 human breast cancer cell lines
• Positive and negative controls established for inhibitor screening
• Secondary validation using commercially available [14C]-labeled fructose
• A [18F]-labeled probe in preparation for clinical evaluation in breast cancer detection via PET
• Plans for complementary fluorescent glucose probes to enable a two-color assay tracking both GLUT5 and GLUT1 activity
• Potential partnership with the Centre for High-Throughput Chemical Biology at Simon Fraser University for access to automated HTS infrastructure

Technology readiness level

The assay has been validated in published work using 12-well culture plates with a small library of potential GLUT5 substrates. Preliminary efforts to translate the method to 96-well plates are underway, with remaining work focused on standardizing cell culturing conditions across well surfaces and configurations. Known GLUT5 inhibitors will be used to optimize reproducibility for HTS compatibility. The [18F]-labeled probe is advancing toward clinical evaluation for breast cancer PET imaging. The technology is at an early-to-mid development stage, with the core concept validated but full HTS scalability and dual-transporter capability still requiring further development and resource investment.