A high-throughput cell-based method to predict the unbound drug fraction in the brain

Area of Science:

• Pharmacology
• Neuroscience
• Biotechnology

Background:

• Optimizing drug efficacy in the brain necessitates understanding local unbound drug exposure.
• Current methods for measuring unbound drug fraction in the brain (fu,brain) require invasive brain tissue sampling.
• This limits early-stage drug discovery and optimization.

Purpose of the Study:

• To develop a novel, rapid, and high-throughput methodology for estimating fu,brain.
• To provide an alternative to animal-based tissue analysis for drug discovery.
• To enable earlier implementation of unbound drug exposure estimations in the drug development pipeline.

Main Methods:

• Utilized homogenates of cultured human embryonic kidney (HEK293) cells.
• Measured drug binding in a small-scale dialysis apparatus.
• Employed drug-cassette technology for simultaneous measurement of multiple compounds.

Main Results:

• The novel cell homogenate method accurately estimated fu,brain.
• The method demonstrated an average error of 1.9-fold compared to traditional methods.
• The approach proved to be a high-throughput and cost-effective alternative.

Conclusions:

• The proposed cell homogenate method is a viable, inexpensive, and accessible alternative for estimating fu,brain.
• This technique facilitates earlier and more frequent unbound drug exposure assessments in drug discovery.
• It supports faster molecular property modifications during early drug development phases.