title: A high throughput solubility assay for drug discovery using microscale shake-flask and rapid UHPLC-UV-CLND quantification.
abstract: The rapid determination of key physical properties of lead compounds is essential to the drug discovery process. Solubility is one of the most important properties since good solubility is needed not only for obtaining reliable in vitro and in vivo assay results in early discovery but also to ensure sufficient concentration of the drug being in circulation to get the desired therapeutic exposure at the target of interest. In order for medicinal chemists to tune solubility of lead compounds, a rapid assay is needed to provide solubility data that is accurate and predictive so that it can be reliably used for designing the next generation of compounds with improved properties. To ensure speed and data quality, we developed a high throughput solubility assay that utilizes a single calibration UHPLC-UV-CLND method and a 24h shake-flask format for rapid quantification. A set of 46 model compounds was used to demonstrate that the method is accurate, reproducible and predictive. Here we present development of the assay, including evaluation of quantification method, filtration membranes, equilibrium times, DMSO concentrations, and buffer conditions. A comparison of thermodynamic solubility results to our high throughput 24h shake-flask solubility assay results is also discussed.
authors: Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080-4990, USA. Electronic address: Lin.baiwei@gene.com.
authors: Baiwei
authors: B
authors: Lin
authors: Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080-4990, USA.
authors: Joseph H
authors: JH
authors: Pease
mesh_terms: D001681:Biological Assay / Q000379:methods; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D055808:Drug Discovery / Q000379:methods*; D008163:Luminescent Measurements / Q000379:methods*; D009584:Nitrogen / Q000737:chemistry*; D004364:Pharmaceutical Preparations / Q000737:chemistry*; D012995:Solubility; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D013816:Thermodynamics
chemical_list: D004364:Pharmaceutical Preparations; D009584:Nitrogen; D004121:Dimethyl Sulfoxide
keywords: CLND—chemiluminescence nitrogen detector; HPLC hyphenated technique; Physicochemical properties; Rapid quantitative analysis; Shake-flask solubility assay