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OSU-03012 (AR-12) PDPK1 inhibitor

Name: OSU-03012 (AR-12)
SKU: S1106
Availability: InStock
Rating: 5 (51 reviews)

Cat.No.S1106

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, showing a 2-fold increase in potency over OSU-02067.

Chemical Structure

Molecular Weight: 460.45

Jump to Mechanism of Action Cell Culture & Working Concentration Solubility Chemical Information, Storage & Stability Specificity

Quality Control

Batch: Purity: 99.14%

99.14

Related Targets	PI3K Akt mTOR AMPK GSK-3 ATM/ATR DNA-PK PTEN PP2A PDK PI4K PIKfyve MELK
Related Products	GSK2334470 BX517 PS-48 PS210 PDPK1 Antibody [K10G23] Phospho-PKA C (Thr197) Antibody [L13C4] PS47 Indoprofen

Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Incubation Time	Activity Description
mouse RAW264.7 cells	Function assay	8 h	Antimicrobial activity against Salmonella enterica serovar Typhimurium ATCC 14028 infected in mouse RAW264.7 cells assessed as inhibition of intracellular bacterial growth after 8 hrs, IC50=0.2 μM
mouse RAW264.7 cells	Cytotoxic assay	24 h	Cytotoxicity against mouse RAW264.7 cells assessed as cell viability after 24 hrs by MTT assay, IC50=10 μM
Click to View More Cell Line Experimental Data

Chemical Information, Storage & Stability

Molecular Weight	460.45	Formula	C₂₆H₁₉F₃N₄O	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	742112-33-0	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
Synonyms	N/A			Smiles	C1=CC=C2C(=C1)C=CC3=C2C=CC(=C3)C4=CC(=NN4C5=CC=C(C=C5)NC(=O)CN)C(F)(F)F

Solubility

In vitro
Batch:

DMSO : 11 mg/mL (23.88 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
=	×	×

Dilution Calculator Molecular Weight Calculator

In vivo Batch:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	0.5% methylcellulose 1 0.2% Tween 80 Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	10.000mg/ml (21.72mM)	Taking the 1 mL working solution as an example, take 10 mg of this product, add it to 1 ml of 0.5% methylcellulose+0.2% Tween 80 clear solution, and mix evenly to make it a uniform suspension. The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

Dosage: mg/kg Average weight of animals: g Dosing volume per animal:μL Number of animals:

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Mechanism of Action

Targets/IC₅₀/Ki	PDPK1 ^[1] (Cell-free assay) 5 μM
In vitro	OSU-03012 (AR-12) induces apoptotic death in PC-3 cells with IC50 of 5 µM and reduces the activity of immunoprecipitated p70^S6K. It completely suppresses 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] This compound promotes cell killing to a greater extent in glioma cells than in nontransformed astrocytes. It 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. Its lethality as a single agent or when combined with signaling modulators is not modified in cells lacking expression of BIM or of BAX/BAK. It promotes the release of cathepsin B from the lysosomal compartment and that of AIF from mitochondria. The lethality of this compound 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] It 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] It inhibits cell growth of hepatocellular carcinoma cell lines including Huh7, Hep3B and HepG2 cells with IC50 values below 1 μM. This compound 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 its treatment. ^[4] A recent study shows that it could enhance the susceptibility of (Bcr)-Abl mutant cell lines to -induced apoptosis. ^[5]
Kinase Assay	PDK-1 Kinase Assay
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 (AR-12), 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.
In vivo	OSU-03012 (AR-12) suppresses tumor growth by 57.59% and increases cleaved LC3 in Huh7 tumor xenografts at 200 mg/kg. ^[4] This compound 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] It is well tolerated and inhibits the growth of HMS-97 schwannoma xenografts by 55% after oral administration. ^[7]
References	[1] https://pubmed.ncbi.nlm.nih.gov/15205346/ [2] https://pubmed.ncbi.nlm.nih.gov/16622074/ [3] https://pubmed.ncbi.nlm.nih.gov/17673571/ [4] https://pubmed.ncbi.nlm.nih.gov/19010909/ [5] https://pubmed.ncbi.nlm.nih.gov/15665113/ [6] https://pubmed.ncbi.nlm.nih.gov/17595327/ [7] https://pubmed.ncbi.nlm.nih.gov/19359162/

Applications

Methods	Biomarkers	Images	PMID
Western blot	p-Akt / Akt		18413750
Growth inhibition assay	Cell viability		18413750

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