A high throughput solubility assay for drug discovery using microscale shake-flask and rapid UHPLC-UV-CLND...

Area of Science:

• Medicinal Chemistry
• Drug Discovery
• Analytical Chemistry

Background:

• Solubility is a critical physicochemical property for drug discovery, impacting assay reliability and therapeutic efficacy.
• Accurate and rapid solubility data is essential for medicinal chemists to optimize lead compounds.
• Existing methods may lack the speed or predictive power needed for efficient lead optimization.

Purpose of the Study:

• To develop and validate a high-throughput solubility assay for rapid quantification of lead compounds.
• To establish a reliable method for generating accurate and predictive solubility data in early drug discovery.
• To optimize assay parameters for speed and data quality.

Main Methods:

• Development of a high-throughput solubility assay utilizing a 24-hour shake-flask format.
• Implementation of a single calibration Ultra-High Performance Liquid Chromatography-Ultraviolet-Charged Aerosol Detection (UHPLC-UV-CLND) method for quantification.
• Evaluation of various parameters including filtration membranes, equilibrium times, DMSO concentrations, and buffer conditions.

Main Results:

• The developed assay demonstrated accuracy, reproducibility, and predictive capability using a set of 46 model compounds.
• Optimization of quantification method, filtration, and incubation conditions ensured assay robustness.
• Comparison of thermodynamic solubility with high-throughput assay results validated the method's performance.

Conclusions:

• The developed high-throughput solubility assay provides rapid, accurate, and predictive data crucial for lead compound optimization.
• This assay facilitates efficient drug design by enabling faster iteration cycles for improved compound properties.
• The method is suitable for routine use in drug discovery programs requiring reliable solubility assessments.