High-Throughput Screening of DPPIV Inhibitors Antagonizing GLP-1 Degradation Using an Enzymatic Activated Fluorescent...

Area of Science:

• Biochemistry
• Enzymology
• Pharmacology

Background:

• Dipeptidyl peptidase IV (DPPIV) is an exopeptidase crucial in various physiological processes.
• DPPIV inhibitors are established treatments for diabetes mellitus (DM).
• There is a need for efficient methods to screen and identify novel DPPIV inhibitors.

Purpose of the Study:

• To design and validate a far-red fluorescent probe, DBX-AP, for real-time DPPIV activity monitoring.
• To develop a visual high-throughput screening (HTS) method for DPPIV inhibitors using DBX-AP.
• To identify novel DPPIV inhibitors from a compound library.

Main Methods:

• Molecular docking simulations and functional characteristics of DPPIV were used to design the DBX-AP probe.
• DBX-AP was employed to establish a visual HTS assay for DPPIV inhibitors.
• In vitro and in vivo studies were conducted to evaluate DPPIV activity and inhibitor efficacy in mice and gut microbiota.

Main Results:

• The DBX-AP probe enabled rapid, selective, and real-time monitoring of DPPIV activity.
• A HTS campaign identified three potent DPPIV inhibitors (K784-2660, 6484-0066, E699-0153).
• The identified inhibitors suppressed DPPIV activity in mouse ileum, reducing GLP-1 degradation, and in gut microbiota.

Conclusions:

• DBX-AP is a valuable tool for evaluating DPPIV activity and discovering novel DPPIV inhibitors.
• The developed HTS method offers a promising approach for identifying new therapeutic agents for diabetes mellitus.
• Targeting DPPIV in both host and gut microbiota presents a potential therapeutic strategy for DM.