Rational Design of an Isoform-Specific Fluorogenic Probe for Human UGT2B7: Enabling Functional Analysis and Inhibitor...

Area of Science:

• Biochemistry
• Pharmacology
• Chemical Biology

Background:

• UDP-glucuronosyltransferase 2B7 (UGT2B7) is crucial for drug metabolism and xenobiotic detoxification.
• Existing methods for monitoring UGT2B7 activity are limited, hindering research and drug development.

Purpose of the Study:

• To develop a novel, sensitive, and specific fluorogenic probe for in situ detection of UGT2B7 activity.
• To utilize the probe for functional imaging and screening of UGT2B7 inhibitors.

Main Methods:

• Integrated strategy combining structure-based virtual screening and biochemical validation.
• Design and synthesis of 1,8-naphthalimide derivatives based on the PET mechanism.
• Development of UPro2B7, a UGT2B7-activatable fluorogenic probe.

Main Results:

• UPro2B7 exhibits high isoform specificity and sensitivity for UGT2B7.
• Enabled effective functional imaging of endogenous UGT2B7 with high spatial resolution and ER colocalization.
• Identified Licoagrochalcone A (Lic A) and Glycycoumarin (Gly) as potent UGT2B7 inhibitors, validated in various biological systems.

Conclusions:

• UPro2B7 is a practical and reliable tool for in situ imaging and inhibitor assessment of UGT2B7 activity.
• The developed integrated strategy is effective for engineering isoform-specific activatable fluorogenic probes.
• This work provides a superior alternative to conventional substrates for screening UGT2B7 inhibitors.