PF-573228

For research use only.

Catalog No.S2013

50 publications

PF-573228 Chemical Structure

CAS No. 869288-64-2

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. PF-573228 induces apoptosis.

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Selleck's PF-573228 has been cited by 50 publications

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Biological Activity

Description 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. PF-573228 induces apoptosis.
Targets
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, PF 573228 (1 μM) with 80% inhibition of FAK phosphorylation fails to inhibit cell growth or apoptosis. Similar treatment of cells with PF-228 resulted in inhibition of serum or FN-directed migration and decreased focal adhesion turnover. [1]
Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
A431 NH7oZZlMcW6jc3WgZZN{[Xl? NXq1[nlThjFyIN88US=> NVnrcJJHTE2VTx?= MVXpcohq[mm2czDGRWsheGixc4Doc5J6dGG2aX;uJJdqfGhiSVO1NEBw\iBzMTDuUS=> NETiN3EyPzN7NUW5OC=>
REF52 NXnveVZwU2mwYYPlJIF{e2G7 MoPmglExKM7:TR?= NXn2fZZzTE2VTx?= MoLUbY5pcWKrdIOgeIhmKHCqb4PwbI9zgWyjdHnvckBw\iCIQVugWJlzOzl5IIfpeIghUUN3MDDv[kB,OTByIH7N NYDBfpNnOTd|OUW1PVQ>
PC3 M1Htb2tqdmG|ZTDhd5NigQ>? MWT+NVAh|ryP NG\YT3FFVVOR NEi3T2hqdmirYnn0d{B1cGVicHjvd5Bpd3K7bHH0bY9vKG:oIF\BT{BVgXJ|OUege4l1cCCLQ{WwJI9nKDFyMDDuUS=> NVnJZYlWOTd|OUW1PVQ>
SKOV-3 MnfuT4lv[XOnIHHzd4F6 MoH5glExKM7:TR?= NHfV[VJFVVOR NHPxOlRqdmirYnn0d{B1cGVicHjvd5Bpd3K7bHH0bY9vKG:oIF\BT{BVgXJ|OUege4l1cCCLQ{WwJI9nKDVyIH7N NVq3RZRXOTd|OUW1PVQ>
L3.6p1 Mny3T4lv[XOnIHHzd4F6 MXf+NVAh|ryP M1TwR2ROW09? MUPpcohq[mm2czD0bIUheGixc4Doc5J6dGG2aX;uJI9nKE[DSzDUfZI{QTdid3n0bEBKSzVyIH;mJFMxOCCwTR?= Mni4NVc{QTV3OUS=
F-G MnXYT4lv[XOnIHHzd4F6 MVr+NVAh|ryP NHXYWVRFVVOR M3f4SolvcGmkaYTzJJRp\SCyaH;zdIhwenmuYYTpc44hd2ZiRlHLJHR6ejN7NzD3bZRpKEmFNUCgc4YhOzBibl2= MV[xO|M6PTV7NB?=
MDCK NV3hOoJjU2mwYYPlJIF{e2G7 NGPpS4Z,OTBizszN NGHFOZpFVVOR MnrDbY5pcWKrdIOgeIhmKHCqb4PwbI9zgWyjdHnvckBw\iCIQVugWJlzOzl5IIfpeIghUUN3MDDv[kA2ODBibl2= M4P4U|E4Ozl3NUm0
PC3 NYHz[XFyT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= NWWyfGJTOTBizszN MXTEUXNQ MVjzbYdvcW[rY3HueIx6KGmwaHnibZR{KGOnbHyg[5Jwf3SqLh?= M{nybFE4Ozl3NUm0
REF52 M2\ybmdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 NVq4WoZWOTBizszN NIKxbnZFVVOR NYfHNJYxe2mpbnnmbYNidnSueTDpcohq[mm2czDj[YxtKGe{b4f0bE4> MX[xO|M6PTV7NB?=
MDCK NV3MdXJHSXCxcITvd4l{KGG|c3H5 NXjxRW06OTBizszN M1fH[GROW09? MVzpcoR2[2W|IHHwc5B1d3Orcx?= NYnQfXlVOTd|OUW1PVQ>
REF52 M13zcGFxd3C2b4Ppd{Bie3OjeR?= NYHCNIJROTBizszN MXnEUXNQ NFmxVoJqdmS3Y3XzJIFxd3C2b4Ppdy=> MVyxO|M6PTV7NB?=
REF52 MnzoSpVv[3Srb36gZZN{[Xl? MYCxNEDPxE1? MmXrSG1UVw>? Mo[4Zoxw[2u|IIPldpVuKGGwZDDGUk1{fGmvdXzheIVlKG2rZ4LheIlwdg>? M2[4OFE4Ozl3NUm0
platelet MljSSpVv[3Srb36gZZN{[Xl? NFH1WpgyKM7:TR?= M1fZd2ROW09? MUjpcohq[mm2czDwcIF1\WyndDDh[4dz\WejdHnvckBidmRic4Dy[YFlcW6p MnnjNVk4OTZ6MEO=
platelet M{j6OWZ2dmO2aX;uJIF{e2G7 NXPUUJZvOSEQvF2= M{izTmROW09? NIi3cJlt\WGmczD0c{BqdmirYnn0bY9vKG:oIGDBT{BidmRiQVvU MlHlNVk4OTZ6MEO=
platelet MX7GeY5kfGmxbjDhd5NigQ>? NYfUdo5bOSEQvF2= M4fNNWROW09? NH;qVFhjdG:la4OgZ4Ft[2m3bTDtc4JqdGm8YYTpc44h[W6mIHTlcpNmKGe{YX71cIUhe2WlcnX0bY9v M{Gx[FE6PzF4OECz
4T1 MYLGeY5kfGmxbjDhd5NigQ>? M374UmROW09? NInMSpNi[m:uaYPo[ZMhfGinIHnueIVz[WO2aX;uJIJmfHenZX6g{tI{KGmwdHXndolvKGGwZDDU{tJTNUmL NV7jZ2tNOTl5NEC0N|M>
MCF7 NGfySXJMcW6jc3WgZZN{[Xl? MnjLglExKM7:TR?= NUDUdlh3TE2VTx?= NVnWdINucW6qaXLpeJMhfGinIIDoc5NxcG:{eXzheIlwdiCxZjDGRWshXHm{M{m3JJdqfGhiSVO1NEBw\iB2M{Cgcm0> NVfvTXgyOjB|NUS3PFA>
TamR NELxfIlMcW6jc3WgZZN{[Xl? M1XLSZ4yOCEQvF2= M4X4TmROW09? NFvpUpdqdmirYnn0d{B1cGVicHjvd5Bpd3K7bHH0bY9vKG:oIF\BT{BVgXJ|OUege4l1cCCLQ{WwJI9nKDVyIH7N NHf3NVUzODN3NEe4NC=>
FasR MlPvT4lv[XOnIHHzd4F6 NGTRWpB,OTBizszN NEPCR3VFVVOR MlHGbY5pcWKrdIOgeIhmKHCqb4PwbI9zgWyjdHnvckBw\iCIQVugWJlzOzl5IIfpeIghUUN3MDDv[kAyOzBibl2= NWrEcZhnOjB|NUS3PFA>
TamR MYXGeY5kfGmxbjDhd5NigQ>? M4LuWlEh|ryP NG\GO3NFVVOR M1;oSIlvcGmkaYTzJINmdGxibXnndoF1cW:w M2S0WVIxOzV2N{iw
FasR NYrzc2VUTnWwY4Tpc44h[XO|YYm= M1q2dFEh|ryP M3z5PWROW09? NIPCUHRqdmirYnn0d{Bk\WyuIH3p[5JifGmxbh?= MlraNlA{PTR5OEC=
endothelial cell MVzLbY5ie2ViYYPzZZk> NXnaXYNEPDBibl2= M3[wW2ROW09? NI\5eI5qdmirYnn0d{BJOk9{LXnu[JVk\WRicHjvd5Bpd3K7bHH0bY9vKG:oIF\BTy=> M3u1SVIyOjF{NECy
endothelial cell M{XDdGZ2dmO2aX;uJIF{e2G7 M2LTOlQxKG6P M4\YSWROW09? M3rZeIlvcGmkaYTzJGgzVzJvaX7keYNm\CC|dILld5Mh\mmkZYKg[o9zdWG2aX;u M3;iXVIyOjF{NECy
endothelial cell MlfORZBweHSxc3nzJIF{e2G7 NF;XNGE1OCCwTR?= NGTqbWhFVVOR NX74OVU1cW6qaXLpeJMh[XCxcITvd4l{ MY[yNVIyOjRyMh?=
GH3 Mk\jSpVv[3Srb36gZZN{[Xl? MU[zJO69VQ>? NF7MSVdFVVOR NHjHOpJqdmO{ZXHz[ZMhUUtqQ3GpJIFueGyrdIXk[S=> MoTQNlE6OjV3MUK=
GH3 MVXGeY5kfGmxbjDhd5NigQ>? NHP4U4g{KM7:TR?= NVXkTllTTE2VTx?= MWPlcohidmOnczDCT2NiNWOqYX7u[Ywh[WO2aY\peJk> NV[xR|JnOjF7MkW1NVI>
HUVEC M3r6WYN6fG:2b4jpZ4l1gSCjc4PhfS=> M3LE[54yOCEQvF2= NFjXTXBFVVOR MUDpcZBicXK|IHXu[I91cGWuaXHsJINmdGxidnnhZoltcXS7 NWTKPHZ[OjJyN{WwOVc>
HUVEC NUf2bWFqU2mwYYPlJIF{e2G7 MVu1JO69VQ>? MWLEUXNQ NIn1XGVqdmirYnn0d{BHSUtia3nuZZNmKGGldHn2bZR6 MmW0NlIxPzVyNUe=
HUVEC MlTtSpVv[3Srb36gZZN{[Xl? NF73R|A2KM7:TR?= NIXyW2NFVVOR MnHvbY5lfWOnczDj[YxtKGO7Y3zlJIFzemW|dB?= NGDKdY8zOjB5NUC1Oy=>
HUVEC M1zMc2Fxd3C2b4Ppd{Bie3OjeR?= MXS1JO69VQ>? MWLEUXNQ MYTpcoR2[2W|IHHwc5B1d3Orcx?= NHfZXlUzOjB5NUC1Oy=>
HUVEC NYnZTohjTnWwY4Tpc44h[XO|YYm= NYPY[Xc1PSEQvF2= NELnOGtFVVOR MYfpcZBm\GW|IHXu[I91cGWuaXHsJINmdGxibXnndoF1cW:wIHHu[EBidHSncoOgeIhmKGOnbHz1cIFzKGGldHnuJIN6fG:|a3Xs[ZRwdg>? MUWyNlA4PTB3Nx?=
HUVEC MVHGeY5kfGmxbjDhd5NigQ>? M1vqfVUh|ryP NYDSfY1ITE2VTx?= NY[0TYpS[myxY3vzJGhWXkWFIIPwdo92fGmwZzDvckBkd2yuYXflckBKKGenbIO= NIXEV2czOjB5NUC1Oy=>
human peripheral blood T cells M{\vXGtqdmG|ZTDhd5NigQ>? NWDOcYN6hjFyIN88US=> M4riPGROW09? NVLWbXlzcW6qaXLpeJMhe2m2ZT3zdIVkcW[rYzDwbI9{eGixconsZZRqd25ib3[gSmFM NF;p[|czOzl{OEG4PC=>
human peripheral blood T cells M3TtOWZ2dmO2aX;uJIF{e2G7 NInFSVV,OTBizszN MmLkSG1UVw>? MnfpbY1x[Wm{czDUR3IucW6mdXPl[EBVKGOnbHygcY9zeGixbH;nbYNidCClaHHu[4V{KGGwZDDhcJRmenNiYXP0bZZqfHlib3[gVohwSQ>? NUHEcGg3OjN7MkixPFg>
human peripheral blood T cells NU\xdpU3TnWwY4Tpc44h[XO|YYm= MojEglExKM7:TR?= M1nyTWROW09? M{HIWIlvcGmkaYTzJJBpd3OyaH;yfYxifGmxbjDv[kBbSVBvN{CgZY5lKEyDVB?= MYKyN|kzQDF6OB?=
human peripheral blood T cells NWO0NYRITnWwY4Tpc44h[XO|YYm= MnO2glExKM7:TR?= MUDEUXNQ NWXJU|hrcW2yYXnyd{BCdnSrZ3XuMYRmeGWwZHXueEBVKGOnbHygZ49vcnWpYYTpc44> MUSyN|kzQDF6OB?=

... Click to View More Cell Line Experimental Data
Assay
Methods Test Index PMID
Western blot
cyclin B1; 

PubMed: 30761269     


(D) On the third day, PF-573228-treated cells were harvested and subjected to Western blot analysis for cyclin B1. Cyclin B1 levels were much higher in A549 cells with 1 μM PF-573228 or without PF-573228 treatment than in the cells treated with higher concentrations of PF-573228. (E) After 10 μM PF-573228 treatment, cyclin B1 levels declined markedly in H460 cells. (F) PF-573228 administration slightly reduced cyclin B1 levels in H1299 cells. 

p-FAK / FAK ; 

PubMed: 30761269     


FAK expression levels and FAK activity, as measured by the phosphorylation of FAK at tyrosine 576 and 577, were quantified by Western blot analysis after treatment of lung cancer cells with PF-573228 for 3 days.

Lamin A / Lamin C; 

PubMed: 30761269     


(B) The cells treated with 10 μM PF-573228 exhibited a decrease in p-FAK levels. Lamin A and lamin C expression levels were much lower in A549 cells exposed to 10 μM PF-573228. 

30761269
Immunofluorescence
FAK / F-actin; 

PubMed: 30761269     


(B) The cells were stained with phalloidin to visualize F-actin (red) and a FAK antibody to visualize the FAK distribution (green). In cells without PF-573228 administration, FAK translocated to focal adhesions at the tips of actin stress fibers, and the focal adhesions were relatively large. When cells were exposed to 10 μM PF-573228, FAK translocation to focal adhesions was reduced, and the sizes of the focal adhesions were smaller. Nuclei in cells treated with PF-573228 were deformed, as visualized with DAPI staining, whereas most nuclei in cells without PF-573228 treatment were oval shaped. 

Emerin; 

PubMed: 30761269     


(A) After PF-573228 treatment of A549 cells, the cells were fixed and stained with phalloidin to label F-actin (red) and an antibody against emerin (green) to outline the nuclear shape. Cells treated with PF-573228 were extremely large and had deformed nuclei, whereas mostly oval-like nuclei were present in the cells without PF-573228 treatment. 

30761269
Growth inhibition assay
Cell viability; 

PubMed: 30761269     


Three different types of lung cancer cells, (A) A549 lung adenocarcinoma and (B) H460, and (C) H1299 large cell carcinoma, were selected for the PF-573228 administration regimen. Cell growth curves of the three lung cancer cell lines treated with various doses of PF-573228 for 4 days were established. The administration of PF-573228 at 10 μM to the lung cancer cells effectively suppressed cell growth in vitro, as proliferative activity totally ceased in the cells exposed to 10 μM PF-573228.

30761269
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 PF-573,228 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].

Protocol

Kinase Assay:

[1]
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Affinity determination:

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
Cell Research:

[1]
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  • Cell lines: REF52 or PC3 cells
  • Concentrations: ~10 μM
  • Incubation Time: 3 days
  • Method:

    Growth assays are performed by seeding 1 × 104 REF52 or PC3 cells/well of a 24-well plate in triplicate 24 h prior to daily treatment with the indicated concentrations of each inhibitor for 3 days. Subsequently, the cells are harvested and counted.


    (Only for Reference)
Animal Research:

[2]
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  • Animal Models: Ctrl-MT and MFCKO-MT mice
  • Dosages: 5 mg/kg
  • Administration: oral administration
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 26 mg/mL (52.9 mM)
Water Insoluble
Ethanol Insoluble
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
5% DMSO+2% Tween 80+30% PEG 300+ddH2O
For best results, use promptly after mixing. 		5mg/mL

* 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.

Chemical Information

Molecular Weight 491.49
Formula

C22H20F3N5O3S
CAS No. 869288-64-2
Storage powder
in 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

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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

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Frequently Asked Questions

  • Question 1:

    Would you please let me know the detail of how to dissolve PF-573228 (Catalog No.S2013) for in vivo study (oral administration)?

  • Answer:

    PF-573228 in 30% PEG400+0.5% Tween80+ 5% Propylene glycol at 30mg/ml is a suspension. If you will use the compound for oral gavage, this suspension is fine for it.

selleck catalog

FAK Signaling Pathway Map

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID