Brain pharmacokinetic studies

The study was approved by the Institutional Animal Care and Use of Committee of Daegu-Gyeongbuk Medical Innovation Foundation (DGMIF) and performed in accordance with protocols approved by the Institutional Animal Care and Use of Committee. Compound 9b and SAHA were intravenously administered to ICR Male mice, aged 7–8 weeks and weighing 25–30 g at 2 mg/kg with solution formulation of 10% DMSO, 70% PEG400, and 20% saline. Brain and plasma samples were collected at 0.5 and 1 h time points. Brain samples were homogenized at a 1:4 ratio of sample weight (g) to PBS volume (mL). Aliquots (20 μL) of brain homogenate were mixed with 180 μL of acetonitrile, vortexed and centrifuged at 15,000 rpm for 5 minutes at 4 °C. The resulting supernatants were used for LC-MS/MS analysis. Mice plasma samples were collected at 0.5 and 1 h time points. 180 μL of acetonitrile was added to 20 μL of plasma samples, vortexed and centrifuged at 15,000 rpm for 5 minutes at 4 °C. Concentrations of compound 9b and SAHA in mice brain and plasma were determined using an LC-MS/MS. The data found (ng/mL) was multiplied by its dilution factors to obtain the concentration (ng/mL) of compound 9b and SAHA in brain and plasma.