Fluorescent cell-based SGLT1 and GLUT5 transporter screening assay for GRAS compound identification

Overview

This offering provides a high-throughput, cell-based screening assay for identifying compounds that modulate the activity of human glucose transporters SGLT1 and GLUT5. The approach combines stable cell lines expressing these transporters with the fluorescent reporter substrate 1-NBD-Glucose, enabling detection of both inhibitors and activators in a standard microplate format. The assay is designed for screening Generally Recognized as Safe (GRAS) compounds, making it well suited for food, nutraceutical, and pharmaceutical ingredient discovery programs seeking to modulate intestinal sugar absorption or transport.

Technical specifications

• Cell-based platform: Stable cell lines expressing human SGLT1 (and potentially GLUT5), with CHO cells as a straightforward option and Caco2 human intestinal cells available for more physiologically relevant validation.
• Fluorescent detection: Uses 1-NBD-Glucose as a reporter probe for real-time measurement of transporter activity, eliminating the need for radiolabeled substrates in primary screening.
• High-throughput format: 96-well microplate configuration enabling efficient screening of large compound libraries.
• Validated performance: Previously validated against a library of over 2,000 compounds, successfully identifying multiple novel natural and synthetic SGLT1 inhibitors and activators across structurally diverse chemical classes.
• Benchmarking: Confirmed activity of known SGLT1 inhibitor phloridzin and identified novel natural inhibitor (+)-pteryxin, with orthogonal validation performed in Caco2 transwell models using radiolabeled substrate.
• Modeling capability: Reaction conditions can be modified to simulate physicochemical properties of the intestinal lumen, such as viscosity, to study environmental effects on transporter activity.
• Extensible scope: Methodology adaptable to GLUT5 screening if a suitable fluorescent probe becomes available.

Technology readiness level

The core assay concept has been validated through peer-reviewed publications demonstrating successful screening of a 2,000+ compound library and identification of novel SGLT1 modulators. The proposed project involves producing new stable transporter-expressing cell lines, developing the optimized fluorescent screening protocol, and executing GRAS compound screening campaigns. Project duration ranges from 1 to 2 years depending on scope, budget, and whether one or both transporters are targeted. The approach is positioned at a stage ready for collaborative development and pilot screening engagements with industry partners.