PF-00562271 Besylate

Catalog No.S2672 Synonyms: PF-562271 Besylate

For research use only.

PF-00562271 Besylate (PF-562271) is the benzenesulfonate salt of PF-562271, which is a potent, ATP-competitive, reversible inhibitor of FAK with IC50 of 1.5 nM, ~10-fold less potent for Pyk2 than FAK and >100-fold selectivity against other protein kinases, except for some CDKs. Phase 1.

PF-00562271 Besylate Chemical Structure

CAS No. 939791-38-5

Selleck's PF-00562271 Besylate has been cited by 22 publications

Purity & Quality Control

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Biological Activity

Description PF-00562271 Besylate (PF-562271) is the benzenesulfonate salt of PF-562271, which is a potent, ATP-competitive, reversible inhibitor of FAK with IC50 of 1.5 nM, ~10-fold less potent for Pyk2 than FAK and >100-fold selectivity against other protein kinases, except for some CDKs. Phase 1.
Targets
FAK [1]
(Cell-free assay)
PYK2 [1]
(Cell-free assay)
CDK2/CyclinE [1]
(Cell-free assay)
CDK3/CyclinE [1]
(Cell-free assay)
CDK1/CyclinB [1]
(Cell-free assay)
Click to View More Targets
1.5 nM 13 nM 30 nM 47 nM 58 nM
In vitro

PF-562271 Besylate shows the selective inhibitory effects on FAK and Pyk2 tyrosine kinase activity with IC50 of 1.5 nM and 14 nM, respectively. And in cell-based assays, the IC50 of PF-562271 is shown to be 5 nM for FAK, which is more selective compared to other kinase targets. [1] In 2 dimensional (2D) cultures, PF-562271 results in a dose-dependent cell proliferation inhibition in FAK WT, FAK−/− and FAK kinase-deficient (KD) cells with IC50 of 3.3 μM, 2.08 μM and 2.01 μM, respectively. [2]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
MV-4-11 cell NXf4[4Z6T3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= MVfJcohq[mm2aX;uJI9nKGi3bXHuJG1XNTRvMUGgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfUwhUUN3ME2wMlI4PjZizszN MoPtV2FPT0WU
human SW982 cell NHPoZWdIem:5dHigbY5pcWKrdHnvckBie3OjeR?= MnnUTY5pcWKrdHnvckBw\iCqdX3hckBUXzl6MjDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyPUCuN|I5OiEQvF2= NGXJfG1USU6JRWK=
human KM12 cell M4PuPGdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 M3Ozb2lvcGmkaYTpc44hd2ZiaIXtZY4hU01zMjDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyPUCuN|g2PTdizszN NWLYb2dsW0GQR1XS
human COLO-205 cell MWjHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? MXHJcohq[mm2aX;uJI9nKGi3bXHuJGNQVE9vMkC1JINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZktKEmFNUC9NE41QDZ3ODFOwG0> NYK5U3pmW0GQR1XS
human COLO-829 cell NETZ[YhIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NVPae3ZPUW6qaXLpeIlwdiCxZjDoeY1idiCFT1zPMVgzQSClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LDDJR|UxRTBwN{[xO|Yh|ryP MoDDV2FPT0WU
human MG-63 cell MY\Hdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? NXKxb49LUW6qaXLpeIlwdiCxZjDoeY1idiCPRz22N{Bk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVAvQDB4M{eg{txO NUTYNHZ{W0GQR1XS
human IGROV-1 cell Mn\tS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? MULJcohq[mm2aX;uJI9nKGi3bXHuJGlIWk:YLUGgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfUwhUUN3ME2wMlgyODN6IN88US=> M{PobXNCVkeHUh?=
human NCI-H650 cell NWPOepJpT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= Mo\GTY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSE[1NEBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVAvQDNzNUSg{txO NVTXPXpwW0GQR1XS
human RT-112 cell M4\VPWdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MX7Jcohq[mm2aX;uJI9nKGi3bXHuJHJVNTFzMjDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyPUCuPVg1PiEQvF2= MnLtV2FPT0WU
human BCPAP cell Mn\4S5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? NGfwZmhKdmirYnn0bY9vKG:oIHj1cYFvKEKFUFHQJINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZktKEmFNUC9NU4xOTJ6ODFOwG0> NHjFeZBUSU6JRWK=
ALL-PO cell M13QXmdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 NV7sfGhkUW6qaXLpeIlwdiCxZjDoeY1idiCDTFytVG8h[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigSxiSVO1NF0yNjBzNUi0JO69VQ>? NHTJb|NUSU6JRWK=
human KYSE-270 cell MYfHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? Ml3mTY5pcWKrdHnvckBw\iCqdX3hckBMYVOHLUK3NEBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVEvODR5MUSg{txO MljCV2FPT0WU
human 8305C cell M1K1bmdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 M13XTmlvcGmkaYTpc44hd2ZiaIXtZY4hQDNyNVOgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfUwhUUN3ME2xMlA6QTB2IN88US=> MnvtV2FPT0WU
NCI-H810 cell NUXYe3FST3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= M1;UdGlvcGmkaYTpc44hd2ZiaIXtZY4hVkOLLVi4NVAh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigSxiSVO1NF0yNjFyN{e2JO69VQ>? MnPZV2FPT0WU
human CAL-33 cell NXfjfFBST3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= MVfJcohq[mm2aX;uJI9nKGi3bXHuJGNCVC1|MzDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyPUGuNVI6OzhizszN MUXTRW5ITVJ?
human AN3-CA cell MYHHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? NEjIUGhKdmirYnn0bY9vKG:oIHj1cYFvKEGQMz3DRUBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVEvOjF6Nkeg{txO NYD1cmlyW0GQR1XS
human NKM-1 cell MWPHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? NUTweIV[UW6qaXLpeIlwdiCxZjDoeY1idiCQS12tNUBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVEvOjd3ME[g{txO MmPMV2FPT0WU
human BPH-1 cell NIHPZ4hIem:5dHigbY5pcWKrdHnvckBie3OjeR?= MoTVTY5pcWKrdHnvckBw\iCqdX3hckBDWEhvMTDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyPUGuNlg4PjZizszN NF3sbXdUSU6JRWK=
human MES-SA cell NHvDe41Iem:5dHigbY5pcWKrdHnvckBie3OjeR?= NGDN[nVKdmirYnn0bY9vKG:oIHj1cYFvKE2HUz3TRUBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVEvOzB4OEKg{txO M3\qXXNCVkeHUh?=
human CAL-62 cell M4n3UWdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 M1zSXWlvcGmkaYTpc44hd2ZiaIXtZY4hS0GOLU[yJINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZktKEmFNUC9NU4{OTlyOTFOwG0> M3[1cnNCVkeHUh?=
human KYSE-150 cell NX3zSm9IT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= MnvXTY5pcWKrdHnvckBw\iCqdX3hckBMYVOHLUG1NEBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVEvOzV{M{[g{txO NY\m[lJ1W0GQR1XS
human SK-UT-1 cell NIfTRZJIem:5dHigbY5pcWKrdHnvckBie3OjeR?= MlHWTY5pcWKrdHnvckBw\iCqdX3hckBUUy2XVD2xJINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZktKEmFNUC9NU41PDZ2NzFOwG0> MW\TRW5ITVJ?
human HUTU-80 cell NIn0WHVIem:5dHigbY5pcWKrdHnvckBie3OjeR?= MluxTY5pcWKrdHnvckBw\iCqdX3hckBJXVSXLUiwJINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZktKEmFNUC9NU41PDh6NjFOwG0> MmiwV2FPT0WU
human SIG-M5 cell NVntNGtiT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= M2nvcGlvcGmkaYTpc44hd2ZiaIXtZY4hW0mJLV21JINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZktKEmFNUC9NU41QDR6NzFOwG0> M2r2XHNCVkeHUh?=
human AGS cell MX7Hdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? NUXZXVA6UW6qaXLpeIlwdiCxZjDoeY1idiCDR2OgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfUwhUUN3ME2xMlUzOTJ2IN88US=> M{HrUnNCVkeHUh?=
human ST486 cell M3fqTGdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 NXT4NmV2UW6qaXLpeIlwdiCxZjDoeY1idiCVVES4OkBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVEvPTN{N{ig{txO MVrTRW5ITVJ?
human HSC-2 cell M3W4NWdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 NWrtOFBuUW6qaXLpeIlwdiCxZjDoeY1idiCKU1OtNkBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVEvPTN7NTFOwG0> M3rtRXNCVkeHUh?=
human BC-1 cell NV\6[JpRT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= MVfJcohq[mm2aX;uJI9nKGi3bXHuJGJENTFiY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeTygTWM2OD1zLk[xOlY1KM7:TR?= Mn7yV2FPT0WU
human CGTH-W-1 cell NXXlOZJ4T3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= NYTTXVhzUW6qaXLpeIlwdiCxZjDoeY1idiCFR2TIMXcuOSClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LDDJR|UxRTFwNkG2O|kh|ryP M2fVdXNCVkeHUh?=
human MZ1-PC cell MWnHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? NI\vdGVKdmirYnn0bY9vKG:oIHj1cYFvKE2cMT3QR{Bk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVEvPjJ|MUKg{txO NW\ydYE3W0GQR1XS
human SW1710 cell NHPCbVRIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NV;ITIN7UW6qaXLpeIlwdiCxZjDoeY1idiCVV{G3NVAh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigSxiSVO1NF0yNjZ{NkK4JO69VQ>? NV7ZXIZyW0GQR1XS
human EW-13 cell NUj6OVlkT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= MYTJcohq[mm2aX;uJI9nKGi3bXHuJGVYNTF|IHPlcIwh\3Kxd4ToJIlvKGFiY3XscEB3cWGkaXzpeJkh[XO|YYmsJGlEPTB;MT62N|Q3PiEQvF2= NW\h[|hiW0GQR1XS
human U251 cell NIP0eFdIem:5dHigbY5pcWKrdHnvckBie3OjeR?= MW\Jcohq[mm2aX;uJI9nKGi3bXHuJHUzPTFiY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeTygTWM2OD1zLke0NFMyKM7:TR?= M4DiOnNCVkeHUh?=
human NCI-H460 cell M{TKR2dzd3e2aDDpcohq[mm2aX;uJIF{e2G7 M3jqTWlvcGmkaYTpc44hd2ZiaIXtZY4hVkOLLVi0OlAh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigSxiSVO1NF0zNjB2OEO5JO69VQ>? MnXKV2FPT0WU
human DU-4475 cell NFXnW4FIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NYXFS5RFUW6qaXLpeIlwdiCxZjDoeY1idiCGVT20OFc2KGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XluIFnDOVA:Oi5zNEe1PUDPxE1? NEizV29USU6JRWK=
human MFE-296 cell NEjzVYhIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NVrzTZBLUW6qaXLpeIlwdiCxZjDoeY1idiCPRlWtNlk3KGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XluIFnDOVA:Oi52N{e5NkDPxE1? NXP0XlNyW0GQR1XS
human DU-145 cell M1HFfWdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 NXrISFV7UW6qaXLpeIlwdiCxZjDoeY1idiCGVT2xOFUh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigSxiSVO1NF0zNjR7MUG4JO69VQ>? NYr3VnNHW0GQR1XS
human MDA-MB-231 cell M2j0Xmdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MoHRTY5pcWKrdHnvckBw\iCqdX3hckBOTEFvTVKtNlMyKGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XluIFnDOVA:Oi52OUW3NkDPxE1? NXTFUZVZW0GQR1XS
human SNU-387 cell MWfHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? M1zybmlvcGmkaYTpc44hd2ZiaIXtZY4hW06XLUO4O{Bk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVIvPTJ6MjFOwG0> M4\KbHNCVkeHUh?=
In vivo In several human s.c. xenograft models, PF-562271 exhibits dose-dependent tumor growth inhibition, and produces maximum tumor inhibition for PC-3M, BT474, BxPc3, and LoVo ranging from 78% to 94% inhibition at doses of 25 to 50 mg/kg twice daily, without weight loss, morbidity, or death. [1] PF-562271 (25 mg/kg by p.o.) leads to a significant decrease in tumor progression in both subcutaneous and bone metastasis PC3M-luc-C6 xenograft models. [3] In a Huh7.5 hepatocellular carcinoma xenograft model, combination therapy of sunitinib and PF-562271 targets angiogenesis and tumor aggressiveness, and produces more significant anti-tumor effect than single agent by blocking tumor growth and impacting the ability of the tumor to recover upon withdrawal of the therapy. [4]

Protocol (from reference)

Kinase Assay:

[1]

  • Recombinant kinase assay and enzyme kinetics :

    Briefly, purified-activated FAK kinase domain (amino acid 410–689) is reacted with 50 μM ATP and 10 μg per well of a random peptide polymer of Glu and Tyr, p(Glu/Tyr), in kinase buffer [50 mM HEPES (pH 7.5), 125 mM NaCl, and 48 mM MgCl2] for 15 minutes. Phosphorylation of p(Glu/Tyr) is challenged with serially diluted PF-562271 at 1/2-Log concentrations starting at a top concentration of 1 μM. Each concentration is tested in triplicate. Phosphorylation of p(Glu/Tyr) is detected with a general antiphospho-tyrosine (PY20) antibody followed by horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG antibody. HRP substrate is added, and absorbance readings at 450 nm are obtained after addition of stop solution (2 M H2SO4). IC50 values are determined using the Hill-Slope Model. Broad kinase selectivity profiling is performed in house and by using the KinaseProfiler Selectivity Screening Service available through UpState Biotechnology.

Cell Research:

[2]

  • Cell lines: Squamous cell carcinoma (SCC)
  • Concentrations: 0 to 1 μM
  • Incubation Time: 72 hours
  • Method:

    Cells are plated for 48 hours before addition of PF-562271. After 3 days cells are fixed by addition of ice cold 25% trichloroacetic acid (TCA) solution prior to staining with Sulforhodamine B (SRB) dye solution. Plates are washed with 1% glacial acetic acid, air-dried and resuspended in 10 mM Tris buffer, pH 10.5 before reading absorbance at 540 nm. Curve fitting and generation of IC50 values is carried out using GraphPad Prism 4 software from six replicates.

Animal Research:

[1]

  • Animal Models: PC-3M, BT474, BxPc3, LoVo, U87MG, H125 and H460 cells are injected s.c. into the right flank of athymic female mice .
  • Dosages: ≤100 mg/kg
  • Administration: Administered via p.o.

Solubility (25°C)

In vitro

DMSO 14 mg/mL warmed
(21.03 mM)
Water Insoluble
Ethanol Insoluble

In vivo

Add solvents to the product individually and in order
(Data is from Selleck tests instead of citations):
4% DMSO+30% PEG 300+ddH2O
For best results, use promptly after mixing.

3mg/mL

Chemical Information

Molecular Weight 665.66
Formula

C21H20F3N7O3S.C6H6O3S

CAS No. 939791-38-5
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles CN(C1=C(C=CC=N1)CNC2=NC(=NC=C2C(F)(F)F)NC3=CC4=C(C=C3)NC(=O)C4)S(=O)(=O)C.C1=CC=C(C=C1)S(=O)(=O)O

In vivo Formulation Calculator (Clear solution)

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

mg/kg g μL

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.

Molarity Calculator

Mass Concentration Volume Molecular Weight

Clinical Trial Information

NCT Number Recruitment Interventions Conditions Sponsor/Collaborators Start Date Phases
NCT00666926 Completed Drug: PF00562271 Head and Neck Neoplasm|Prostatic Neoplasm|Pancreatic Neoplasm Verastem Inc. December 2005 Phase 1

(data from https://clinicaltrials.gov, updated on 2022-01-17)

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.

Handling Instructions

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Frequently Asked Questions

Question 1:
We are planning both in vitro and in vivo experiments and want to know how to reconstitute the drug for these purposes?

Answer:
PF-00562271 has poor solubility in DMSO and water. Its solubility in DMSO is only 0.4mg/ml. In a previous literature report (http://www.ncbi.nlm.nih.gov/pubmed/18339875), the author used 5% Gelucire to formulate the compound. You can also consider other co-solvents such as PEG400, CMC, Tween80, and Captisol.

Question 2:
Can you provide with a few common vehicles for PF-00562271, S2672 for use as oral gavage?

Answer:
S2672 PF-00562271 can be dissolved in 0.5% CMC Na at 30 mg/ml as a suspension. If 4% DMSO can be used in your experiment, it will help dissolving the suspension more homogeneously.

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