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PF-573228 FAK inhibitor

Name: PF-573228
SKU: S2013
Availability: InStock
Rating: 5 (98 reviews)

Cat.No.S2013

PF-573228 is an ATP-competitive inhibitor of FAK with IC50 of 4 nM in a cell-free assay, ~50- to 250-fold selective for FAK than Pyk2, CDK1/7 and GSK-3β. This compound induces apoptosis.

Chemical Structure

Molecular Weight: 491.49

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

Quality Control

Batch: Purity: 99.92%

99.92

Related Targets	EGFR VEGFR JAK PDGFR FGFR Src FLT3 HER2 Bcr-Abl HIF BTK ALK Syk VDA
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Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Formulation	Activity Description	PMID
A431	Kinase assay	~10 μM	DMSO	inhibits FAK phosphorylation with IC50 of 11 nM	17395594
REF52	Kinase assay	~10 μM	DMSO	inhibits the phosphorylation of FAK Tyr397 with IC50 of ~100 nM	17395594
PC3	Kinase assay	~10 μM	DMSO	inhibits the phosphorylation of FAK Tyr397 with IC50 of 100 nM	17395594
SKOV-3	Kinase assay	~10 μM	DMSO	inhibits the phosphorylation of FAK Tyr397 with IC50 of 50 nM	17395594
L3.6p1	Kinase assay	~10 μM	DMSO	inhibits the phosphorylation of FAK Tyr397 with IC50 of 300 nM	17395594
F-G	Kinase assay	~10 μM	DMSO	inhibits the phosphorylation of FAK Tyr397 with IC50 of 30 nM	17395594
MDCK	Kinase assay	~10 μM	DMSO	inhibits the phosphorylation of FAK Tyr397 with IC50 of 500 nM	17395594
PC3	Growth inhibitory assay	10 μM	DMSO	significantly inhibits cell growth.	17395594
REF52	Growth inhibitory assay	10 μM	DMSO	significantly inhibits cell growth.	17395594
MDCK	Apoptosis assay	10 μM	DMSO	induces apoptosis	17395594
REF52	Apoptosis assay	10 μM	DMSO	induces apoptosis	17395594
REF52	Function assay	10 μM	DMSO	blocks serum and FN-stimulated migration	17395594
platelet	Function assay	1 μM	DMSO	inhibits platelet aggregation and spreading	19716803
platelet	Function assay	1 μM	DMSO	leads to inhibition of PAK and AKT	19716803
platelet	Function assay	1 μM	DMSO	blocks calcium mobilization and dense granule secretion	19716803
4T1	Function assay		DMSO	abolishes the interaction between β3 integrin and TβR-II	19740433
MCF7	Kinase assay	~10 μM	DMSO	inhibits the phosphorylation of FAK Tyr397 with IC50 of 430 nM	20354780
TamR	Kinase assay	~10 μM	DMSO	inhibits the phosphorylation of FAK Tyr397 with IC50 of 50 nM	20354780
FasR	Kinase assay	~10 μM	DMSO	inhibits the phosphorylation of FAK Tyr397 with IC50 of 130 nM	20354780
TamR	Function assay	1 μM	DMSO	inhibits cell migration	20354780
FasR	Function assay	1 μM	DMSO	inhibits cell migration	20354780
endothelial cell	Kinase assay	40 nM	DMSO	inhibits H2O2-induced phosphorylation of FAK	21212402
endothelial cell	Function assay	40 nM	DMSO	inhibits H2O2-induced stress fiber formation	21212402
endothelial cell	Apoptosis assay	40 nM	DMSO	inhibits apoptosis	21212402
GH3	Function assay	3 μM	DMSO	increases IK(Ca) amplitude	21925512
GH3	Function assay	3 μM	DMSO	enhances BKCa-channel activity	21925512
HUVEC	cytotoxicity assay	~10 μM	DMSO	impairs endothelial cell viability	22075057
HUVEC	Kinase assay	5 μM	DMSO	inhibits FAK kinase activity	22075057
HUVEC	Function assay	5 μM	DMSO	induces cell cycle arrest	22075057
HUVEC	Apoptosis assay	5 μM	DMSO	induces apoptosis	22075057
HUVEC	Function assay	5 μM	DMSO	impedes endothelial cell migration and alters the cellular actin cytoskeleton	22075057
HUVEC	Function assay	5 μM	DMSO	blocks HUVEC sprouting on collagen I gels	22075057
human peripheral blood T cells	Kinase assay	~10 μM	DMSO	inhibits site-specific phosphorylation of FAK	23928188
human peripheral blood T cells	Function assay	~10 μM	DMSO	impairs TCR-induced T cell morphological changes and alters activity of RhoA	23928188
human peripheral blood T cells	Function assay	~10 μM	DMSO	inhibits phosphorylation of ZAP-70 and LAT	23928188
human peripheral blood T cells	Function assay	~10 μM	DMSO	impairs Antigen-dependent T cell conjugation	23928188
U-2 OS	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for U-2 OS cells	29435139
A673	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells	29435139
Saos-2	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Saos-2 cells	29435139
BT-37	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-37 cells	29435139
RD	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for RD cells	29435139
SK-N-SH	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-SH cells	29435139
MG 63 (6-TG R)	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for MG 63 (6-TG R) cells	29435139
NB1643	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB1643 cells	29435139
OHS-50	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for OHS-50 cells	29435139
SJ-GBM2	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SJ-GBM2 cells	29435139
SK-N-MC	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells	29435139
NB-EBc1	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB-EBc1 cells	29435139
LAN-5	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for LAN-5 cells	29435139
Click to View More Cell Line Experimental Data

Chemical Information, Storage & Stability

Molecular Weight	491.49	Formula	C₂₂H₂₀F₃N₅O₃S	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	869288-64-2	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
Synonyms	N/A			Smiles	CS(=O)(=O)C1=CC=CC(=C1)CNC2=NC(=NC=C2C(F)(F)F)NC3=CC4=C(C=C3)NC(=O)CC4

Solubility

In vitro
Batch:

DMSO : 26 mg/mL (52.9 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	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.
Clear solution	5%DMSO 1 Corn oil Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	1.640mg/ml (3.34mM)	Taking the 1 mL working solution as an example, add 50 μL of 32.7 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. 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.
Clear solution	5%DMSO 1 2%Tween80 2 30%PEG300 3 63%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	5.000mg/ml (10.17mM)	Taking the 1 mL working solution as an example, add 50 μL of 100 mg/ml clarified DMSO stock solution to 20 μL of Tween80, mix evenly to clarify it; add 300 μL PEG 300 to the above system, mix evenly to clarify; then continue to add 630 μL ddH2O to adjust the volume. to 1 mL. The mixed solution should be used immediately for optimal results.

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	FAK ^[1] (Cell-free assay) 4 nM
In vitro	PF 573228 blocks the phosphorylation of FAK Tyr³⁹⁷ in REF52 cells, PC3 cells, SKOV-3 cells, L3.6p1 and F-G, MDCK cells with IC50 of 30-500 nM. However, this compound (1 μM) with 80% inhibition of FAK phosphorylation fails to inhibit cell growth or apoptosis. Similar treatment of cells with this chemical resulted in inhibition of serum or FN-directed migration and decreased focal adhesion turnover. ^[1]
Kinase Assay	Affinity determination
Kinase Assay	Purified activated FAK kinase domain (amino acids 410–689) is reacted with 50 μM ATP, and 10 μg/well of a random peptide polymer of Glu and Tyr (molar ratio of 4:1), poly(Glu/Tyr) in kinase buffer (50 mM HEPES, pH 7.5, 125 mM NaCl, 48 mM MgCl₂) for 15 min. Phosphorylation of poly(Glu/Tyr) is challenged with serially diluted compounds at 1/2-Log concentrations starting at a top concentration of 1 μM. Each concentration is run in triplicate. Phosphorylation of poly(Glu/Tyr) is detected with a general anti-phospho-tyrosine (PY20) antibody, followed by horseradish peroxidase-conjugated goat anti-mouse IgG antibody. The standard horseradish peroxidase substrate 3, 3
In vivo	Inhibition of FAK by PF-573,228 in Ctrl-MT mice leads to a significant suppression of mammary tumorigenesis as well as lung metastasis. In contrast, treatment of MFCKO-MT mice with this compound did not affect the initiation of mammary tumors in these mice, as would be expected due to the absence of FAK in mammary epithelial cells of these mice ^[2].
References	[1] http://www.ncbi.nlm.nih.gov/pubmed?term=17395594 [2] https://pubmed.ncbi.nlm.nih.gov/21471206/

Applications

Methods	Biomarkers	PMID
Western blot	Lamin A / Lamin C p-FAK / FAK cyclin B1	30761269
Immunofluorescence	Emerin FAK / F-actin	30761269
Growth inhibition assay	Cell viability	30761269

Tech Support

Handling Instructions

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Frequently Asked Questions

Question 1:
Would you please let me know the detail of how to dissolve it for in vivo study (oral administration)? This compound is referred to as Catalog No.S2013.

Answer:
A suspension of this compound in 30% PEG400+0.5% Tween80+ 5% Propylene glycol at 30mg/ml is fine for oral gavage.

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