High-throughput screening of LogD by using a sample pooling approach based on the traditional shake flask method

Area of Science:

• Pharmacokinetics and Drug Metabolism
• Analytical Chemistry
• Medicinal Chemistry

Background:

• Accurate determination of lipophilicity (LogD or LogP) is crucial for drug discovery.
• Traditional methods for LogD screening can be time-consuming and sample-intensive.
• High-throughput screening (HTS) requires efficient and reliable analytical techniques.

Purpose of the Study:

• To develop and validate a new high-throughput method for LogD/LogP screening.
• To enable simultaneous measurement of multiple compounds using sample pooling.
• To reduce the number of samples required for bioanalysis in early drug discovery.

Main Methods:

• Integration of the shake flask method with rapid generic LC-MS/MS bioanalysis.
• Implementation of a sample pooling strategy for simultaneous compound analysis.
• Validation using structurally diverse compounds with a wide LogD range (-0.04 to 6.01), including drug standards and new chemical entities.

Main Results:

• High correlation (RMSE = 0.21, R² = 0.9879) between single and pooled compound LogD measurements.
• Demonstrated accuracy for simultaneous measurement of at least 37 compounds.
• Established tolerance for up to 0.5% DMSO in the assay, with significant reduction in bioanalysis samples compared to conventional methods.

Conclusions:

• The developed method offers a robust and efficient approach for high-throughput LogD/LogP screening.
• Sample pooling significantly enhances throughput and reduces analytical burden in drug discovery.
• This advancement facilitates faster assessment of lipophilicity for drug candidates, streamlining the discovery process.