2-amino-N-{4-[5-(phenanthren-2-yl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]phenyl}acetamide

• ChemBase ID: 72514
• Molecular Formular: C26H19F3N4O
• Molecular Mass: 460.4504696
• Monoisotopic Mass: 460.15109591
• SMILES: c12c(ccc3c1ccc(c3)c1cc(nn1c1ccc(cc1)NC(=O)CN)C(F)(F)F)cccc2
• Canonical SMILES: NCC(=O)Nc1ccc(cc1)n1nc(cc1c1ccc2c(c1)ccc1c2cccc1)C(F)(F)F
• InChI: InChI=1S/C26H19F3N4O/c27-26(28,29)24-14-23(33(32-24)20-10-8-19(9-11-20)31-25(34)15-30)18-7-12-22-17(13-18)6-5-16-3-1-2-4-21(16)22/h1-14H,15,30H2,(H,31,34)
• InChIKey: YULUCECVQOCQFQ-UHFFFAOYSA-N

NAMES AND DATABASE IDS

Names

• IUPAC name: 2-amino-N-{4-[5-(phenanthren-2-yl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]phenyl}acetamide
• IUPAC Traditional name: 2-amino-N-{4-[5-(phenanthren-2-yl)-3-(trifluoromethyl)pyrazol-1-yl]phenyl}acetamide
• Synonyms: AR-12; OSU-03012

Database IDs

• CAS Number: 742112-33-0
• PubChem SID: 162037439
• PubChem CID: 10027278

CALCULATED PROPERTIES

• Acid pKa: 14.522442
• H Acceptors: 3
• H Donor: 2
• LogD (pH = 5.5): 2.8094633
• LogD (pH = 7.4): 4.5038776
• Log P: 5.1825237
• Molar Refractivity: 126.1582 cm^3
• Polarizability: 50.648296 Å^3
• Polar Surface Area: 72.94 Å^2
• Rotatable Bonds: 5
• Lipinski's Rule of Five: false

PROPERTIES

Physical Property

• Solubility: DMSO

Safety Information

• Storage Condition: -20°C

Pharmacology Properties

• Target: PDK

Product Information

• Salt Data: Free Base

DETAILS

Selleck Chemicals - S1106

Research Area

• Description: Cancer

Biological Activity

• Description: OSU-03012 is a potent inhibitor of recombinant PDK-1 with IC50 of 5 μM.
• Targets: PDK-1
• IC50: 5 μM ^[1]
• In Vitro: OSU-03012 induces apoptotic death in PC-3 cells with IC50 of 5 μM and reduces the activity of immunoprecipitated p70^S6K. OSU-03012 completely suppress cell growth in a diverse range of tumor cell lines at concentrations of 3–5 μm, as compared with the concentration of at least 50 μm required for celecoxib. ^[1] OSU-03012 promotes cell killing to a greater extent in glioma cells than in nontransformed astrocytes. OSU-03012 causes a dose-dependent induction of cell death that is not altered by p53 mutation, expression of ERBB1 VIII, or loss of phosphatase and tensin function due to a homolog deletion on chromosome 10. OSU-03012 and ionizing radiation cause an additive, caspase-independent elevation in cell killing. OSU-03012 lethality as a single agent or when combined with signaling modulators is not modified in cells lacking expression of BIM or of BAX/BAK. OSU-03012 promotes the release of cathepsin B from the lysosomal compartment and that of AIF from mitochondria. The lethality of OSU-03012 is attenuated in protein kinase R-like endoplasmic reticulum kinase-/- cells, which correlated with the reduced cleavage of BID and suppression of cathepsin B and AIF release into the cytosol. ^[2] OSU-03012 inhibits thyroid cancer cell (NPA, WRO, and ARO cells) proliferation, migration and induces apoptosis, which results in an increase of cells in the S phase without an increase of cells in G2. OSU-03012 is an ATP-competitive inhibitor of PAK activity and suppresses the phosphorylation of AKT in thyroid cancer cells. ^[3] OSU-03012 inhibits cell growth of hepatocellular carcinoma cell lines including Huh7, Hep3B and HepG2 cells with IC50 values below 1 μM. OSU-03012 does not suppress PDK1 or AKT activity or induce cellular apoptosis but induces autophagy in Huh7 cells. Moreover, accumulation of reactive oxygen species (ROS) is detected after OSU-03012 treatment. ^[4] A recent study shows that OSU-03012 could enhance the susceptibility of (Bcr)-Abl mutant cell lines to imatinib-induced apoptosis. ^[5]
• In Vivo: OSU-03012 suppresses tumor growth by 57.59% and increases cleaved LC3 in Huh7 tumor xenografts at 200 mg/kg. ^[4] OSU-03012 remarkably decreases expression of EGFR protein in the tumors by 48% compared with vehicle controls and also prevents YB-1 from binding to the EGFR promoter in MDA-MB-435/LCC6 xenografts. ^[6] OSU-03012 is well tolerated and inhibits the growth of HMS-97 schwannoma xenografts by 55% after oral administration. ^[7]
• Features: OSU-03012 is a derivative of celecoxib and shows ten-fold greater antitumor activity than celecoxib but it lacks its COX2 inhibitory activity.

Protocol

• Protocol: Kinase Assay ^[1]
• PDK-1 Kinase Assay: This in vitro assay is performed using a PDK-1 kinase assay kit. This cell-free assay is based on the ability of recombinant PDK-1, in the presence of DMSO vehicle or OSU-03012, to activate its downstream serum- and glucocorticoid-regulated kinase which, in turn, phosphorylates the Akt/serum- and glucocorticoid-regulated kinase-specific peptide substrate RPRAATF with [γ-^32P]ATP. The ^32P-phosphorylated peptide substrate is then separated from the residual [γ-^32P]-ATP by using P81 phosphocellulose paper and quantitated in a scintillation counter after three washes with 0.75% phosphoric acid.
• Protocol: Cell Assay ^[1]
• Cell Lines: PC-3 cells
• Concentrations: 0-10 μM
• Incubation Time: ~72 hours
• Methods: The effect of OSU-03012 on PC-3 cell viability is assessed by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay in six replicates. Cells are grown in 10% FBS- supplemented RPMI 1640 in 96-well, flat-bottomed plates for 24 hours. They are exposed to various concentrations of OSU-03012 (0-10 μM) dissolved in DMSO (final concentration ≤0.1%) in 1% serum-containing RPMI 1640 for different time intervals (~72 hours). Controls receive DMSO vehicle at a concentration equal to that in OSU-03012-treated cells. The medium is removed and replaced by 200 μL of 0.5 mg/mL 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide in 10% FBS-containing RPMI 1640. The cells are incubated in the CO₂ incubator at 37 °C for 2 hours. Supernatants are removed from the wells, and the reduced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide dye is solubilized in 200 μL DMSO per well. Absorbance at 570 nm is determined by using a plate reader.
• Protocol: Animal Study ^[4]
• Animal Models: Huh7 tumor xenografts in male BALB/c nude mice
• Formulation: Dissolved in 0.5% methylcellulose, 0.1% Tween 80
• Doses: 100-200 mg/kg
• Administration: Daily by gavage

References

• References: [1] Zhu J, et al. Cancer Res, 2004, 64(12), 4309-4318.
• References: [2] Yacoub A, et al. Mol Pharmacol, 2006, 70(2), 589-603.
• References: [3] Porchia LM, et al. Mol Pharmacol, 2007, 72(5), 1124-1131.
• References: [4] Gao M, et al. Cancer Res, 2008, 68(22), 9348-9357.
• References: [5] Tseng PH, et al. Blood, 2005, 105(10), 4021-4027.
• References: [6] To K, et al. Mol Pharmacol, 2007, 72(3), 641-652.
• References: [7] Lee TX, et al. Eur J Cancer, 2009, 45(9), 1709-1720.

REFERENCES

• Gao M, et al. Cancer Res, 2008, 68(22), 9348-9357.
• To K, et al. Mol Pharmacol, 2007, 72(3), 641-652.
• Zhu J, et al. Cancer Res, 2004, 64(12), 4309-4318.
• Yacoub A, et al. Mol Pharmacol, 2006, 70(2), 589-603.
• Porchia LM, et al. Mol Pharmacol, 2007, 72(5), 1124-1131.
• Tseng PH, et al. Blood, 2005, 105(10), 4021-4027.
• Lee TX, et al. Eur J Cancer, 2009, 45(9), 1709-1720.