OSU-03012 (AR-12)

Catalog No.S1106 Batch:S110601

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Technical Data

Formula

C26H19F3N4O

Molecular Weight 460.45 CAS No. 742112-33-0
Solubility (25°C)* In vitro DMSO 92 mg/mL (199.8 mM)
Ethanol 8 mg/mL (17.37 mM)
Water Insoluble
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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Biological Activity

Description OSU-03012 (AR-12) is a potent inhibitor of recombinant PDK-1(phosphoinositide-dependent kinase 1) with IC50 of 5 μM in a cell-free assay and 2-fold increase in potency over OSU-02067.
Targets
PDPK1 [1]
(Cell-free assay)
5 μM
In vitro

OSU-03012 induces apoptotic death in PC-3 cells with IC50 of 5 µM and reduces the activity of immunoprecipitated p70S6K. 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. [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 -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]

Protocol (from reference)

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.

Cell Assay:

[1]

  • Cell lines

    PC-3 cells

  • Concentrations

    0-10 μM

  • Incubation Time

    ~72 hours

  • Method

    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.

Animal Study:

[4]

  • Animal Models

    Huh7 tumor xenografts in male BALB/c nude mice

  • Dosages

    100-200 mg/kg

  • Administration

    Daily by gavage

Customer Product Validation

Data from [Data independently produced by J Biol Chem, 2014, 289(38), 26155-66]

Data from [Data independently produced by PLoS One, 2013, 8, e75100]

Data from [Anticancer Drugs, 2013, 24(7), 690-8]

Data from [Data independently produced by , , Cancer Medicine, 2017, 6(11):2646-2659]

Selleck's OSU-03012 (AR-12) has been cited by 51 publications

NAC1 confines virus-specific memory formation of CD4+ T cells through the ROCK1-mediated pathway [ J Med Virol, 2023, 95(7):e28957] PubMed: 37465969
Salmonella effector SopF regulates PANoptosis of intestinal epithelial cells to aggravate systemic infection [ Gut Microbes, 2023, 15(1):2180315] PubMed: 36803521
AR12 increases BAG3 expression which is essential for Tau and APP degradation via LC3-associated phagocytosis and macroautophagy [ Aging (Albany NY), 2022, 14(20):8221-8242] PubMed: 36227739
Pulmonary fibrosis distal airway epithelia are dynamically and structurally dysfunctional [ Nat Commun, 2021, 12(1):4566] PubMed: 34315881
Development of potent dual PDK1/AurA kinase inhibitors for cancer therapy: Lead-optimization, structural insights, and ADME-Tox profile [ Eur J Med Chem, 2021, 226:113895] PubMed: 34624821
Decanoic Acid Stimulates Autophagy in D. discoideum [ Cells, 2021, 10(11)2946] PubMed: 34831171
Inhibition of heat shock proteins increases autophagosome formation, and reduces the expression of APP, Tau, SOD1 G93A and TDP-43 [ Aging (Albany NY), 2021, 13(13):17097-17117] PubMed: 34252884
OSU-03012 Disrupts Akt Signaling and Prevents Endometrial Carcinoma Progression in vitro and in vivo [ Drug Des Devel Ther, 2021, 15:1797-1810] PubMed: 33958857
Metabolic Modifier Screen Reveals Secondary Targets of Protein Kinase Inhibitors Within Nucleotide Metabolism [ Cell Chem Biol, 2020, 20;27(2):197-205.e6] PubMed: 31734178
A facile and sensitive method of quantifying glutaminase binding to its inhibitor CB-839 in tissues [ J Genet Genomics, 2020, 47(7):389-395] PubMed: 33004309

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