Development of a high throughput equilibrium solubility assay using miniaturized shake‐flask method in early drug discovery

Retinoic-acid-orphan-receptor-C inhibition suppresses Th17 cells and induces thymic aberrations

“…Microsomal intrinsic clearance was determined by the rate of drug depletion after incubation at 1 μM in male Sprague-Dawley rat liver microsomes (32), plasma protein binding was determined using rapid equilibrium dialysis (33), and solubility was measured according to previously described protocols (34). TR-FRET assay.…”

Physicochemical Profiling in Drug Research and Development

“…The throughput is just medium, as 20 compounds/week can be measured. The MSF method was further developed for HT measurements by Zhou et al [92] where a 96-well plate is used as the source of the samples and DMSO stocks were evaporated via a GeneVac evaporator.…”

Recent Advances in Physicochemical and ADMET Profiling in Drug Discovery

“…For instance, kinetic solubility, albeit simpler and feasible in HT format, fails to properly project the solubility impact on in vivo exposure. Other approaches involving (0.5 -5%) DMSO in the streamlined HT-solubility process may also lead to substantially overestimated equilibrium solubility [48]. In contrast, kinetic solubility derived at the reduced incubation time (e.g.…”

“…In 2007, Zhou et al [48] reported a true HT and miniaturized equilibrium solubility approach utilizing mini-prep vials and fast HPLC. This novel approach addressed most of the caveats encountered by the kinetic or semiequilibrium solubility approaches, thereby exhibiting an excellent agreement with the conventional shake-flask approach for not only commercial drugs but also exigent NCEs in early drug discovery.…”

“…The conventional thermodynamic solubility protocol starting with solid material, also referred to as saturation shake-flask method, reflects not only the molecular interactions of NCEs in solution phase but also the solid packing insights (polymorph state) [5] [45] [46]. The assay typically involves a long (24 -72 h) agitated incubation of NCE powders in a designated buffer at a specific temperature, followed by phase separation (of saturated solution from solute) and quantification [47] [48]. This method is considered the gold standard for solubility determination by industry and US FDA, and is widely utilized to project the in vivo GI absorption [49] and bioavailability of NCEs together with in vitro permeability and metabolic clearance data.…”