OSU-03012  (Synonyms: AR-12; PDK1 inhibitor AR-12)

PC-3 (p53^-/-) human androgen-nonresponsive prostate cancer cells are cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS) at 37°C in a humidified incubator containing 5% CO₂. The effect of Celecoxib and its derivatives (e.g., OSU-03012) (2.5 μM, 5 μM, 7.5 μM and 10 μM) on PC-3 cell viability is assessed by using the MTT assay in six replicates. Cells are grown in 10% FBS- supplemented RPMI 1640 in 96-well, flat-bottomed plates for 24 h, and are exposed to various concentrations of Celecoxib derivatives (e.g., OSU-03012) dissolved in DMSO (final concentration ≤0.1%) in 1% serum-containing RPMI 1640 for different time intervals. Controls receive DMSO vehicle at a concentration equal to that in drug-treated cells. The medium is removed, replaced by 200 μL of 0.5 mg/mL of MTT in 10% FBS-containing RPMI 1640, and cells are incubated in the CO₂ incubator at 37°C for 2 h. Supernatants are removed from the wells, and the reduced MTT dye is solubilized in 200 μL/well DMSO. Absorbance at 570 nm is determined on a plate reader^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Mice^[2]
SCID/Rag2m mice (6-8 weeks old, female) are subcutaneously injected with 1×10⁷ MDA-MB-435/LCC6 cells stably transfected with HER-2/neu. Each mouse is inoculated with the cells on the right and left sides of the lower back. A total of eight mice are injected, each harboring two tumors. After 6 weeks, the mice are randomly assigned into groups (vehicle, 0.5% methyl cellulose/0.1% Tween 80, or OSU-03012 at 200 mg/kg/day). Mice are dosed daily for 3 days with either the vehicle or OSU-03012 by oral gavage. On the fourth day, the study is terminated, mice are sacrificed, and the tumors are collected for chromatin immunoprecipitation (ChIP) and protein isolations.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Zhu J, et al. From the cyclooxygenase-2 inhibitor celecoxib to a novel class of 3-phosphoinositide-dependent protein kinase-1 inhibitors. Cancer Res. 2004 Jun 15;64(12):4309-18.  [Content Brief]

  [2]. To K, et al. The phosphoinositide-dependent kinase-1 inhibitor 2-amino-N-[4-[5-(2-phenanthrenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]phenyl]-acetamide (OSU-03012) prevents Y-box binding protein-1 from inducing epidermal growth factor receptor. Mol Pharmacol. 2007 Sep;72(3):641-52.  [Content Brief]

  [3]. Tina X Lee, et al.Growth Inhibitory and Anti-Tumour Activities of OSU-03012, a Novel PDK-1 Inhibitor, on Vestibular Schwannoma and Malignant Schwannoma Cells. Eur J Cancer  [Content Brief]