¹⁸O-Enabled High-Throughput Acyl Glucuronide Stability Assay

Area of Science:

• Biochemistry
• Pharmacology
• Drug Metabolism

Background:

• Acyl glucuronides (AGs) are metabolites of carboxylic acid drugs.
• AGs can form covalent adducts, potentially leading to drug toxicity.
• Assessing AG chemical stability is vital for drug safety.

Purpose of the Study:

• To develop a high-throughput assay for acyl glucuronide stability.
• To eliminate the need for liquid chromatography (LC) method development.
• To facilitate early-stage drug liability assessment.

Main Methods:

• Utilized 18O-incorporation from [18O] water to monitor AG stability.
• Compared 18O-labeling rates with AG migration tendencies.
• Explored in situ AG generation using human hepatic microsomes.

Main Results:

• Faster 18O-incorporation correlated with shorter AG migration half-lives.
• Differential 18O-incorporation levels accurately predicted AG migration tendency.
• In situ generated AGs yielded results comparable to authentic standards.

Conclusions:

• The 18O-labeling assay provides a simplified, high-throughput method for AG stability assessment.
• This assay requires no LC method development or AG reference standards.
• It effectively facilitates early drug discovery by assessing acyl glucuronide liability.