Home Protein Tyrosine Kinase EGFR inhibitor - HER2 inhibitor - Apoptosis related activator Dacomitinib (PF-00299804)

Dacomitinib (PF-00299804)

Catalog No.S2727 Synonyms: PF299804,PF299

For research use only.

Dacomitinib (PF299804, PF299) is a potent, irreversible pan-ErbB inhibitor, mostly to EGFR with IC50 of 6 nM in a cell-free assay. Dacomitinib inhibits ERBB2 and ERBB4 with IC50 of 45.7 nM and 73.7 nM, respectively. Dacomitinib is effective against NSCLCs with EGFR or ERBB2 mutations (resistant to gefitinib) as well as those harboring the EGFR T790M mutation. Dacomitinib inhibits cell growth and induces apoptosis. Phase 2.

Dacomitinib (PF-00299804) Chemical Structure

CAS No. 1110813-31-4

Selleck's Dacomitinib (PF-00299804) has been cited by 75 publications

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Biological Activity

Description Dacomitinib (PF299804, PF299) is a potent, irreversible pan-ErbB inhibitor, mostly to EGFR with IC50 of 6 nM in a cell-free assay. Dacomitinib inhibits ERBB2 and ERBB4 with IC50 of 45.7 nM and 73.7 nM, respectively. Dacomitinib is effective against NSCLCs with EGFR or ERBB2 mutations (resistant to gefitinib) as well as those harboring the EGFR T790M mutation. Dacomitinib inhibits cell growth and induces apoptosis. Phase 2.
Targets
EGFR [1]
(Cell-free assay)		 ErbB2 [1]
(Cell-free assay)		 ErbB4 [1]
(Cell-free assay)
6.0 nM 45.7 nM 73.7 nM
In vitro

PF299804 is a specific inhibitor of the ERBB family of kinases. PF299804 inhibits EGFR signaling and induces apoptosis in the EGFR T790M-containing H3255 GR cell line. PF299804 is effective in gefitinib-sensitive and gefitinibresistant NSCLC cell lines. PF299804 inhibits the growth of H3255 and HCC827 cells engineered to express EGFR T790M. PF299804 inhibits EGFR phosphorylation in the presence of the T790M mutation. [1] PF-299804 is believed to irreversibly inhibit ERBB tyrosine kinase activity through binding at the ATP site and covalent modification of nucleophilic cysteine residues in the catalytic domains of ERBB family members. [2] PF299804 shows significant growth-inhibitory effects in HER2-amplified gastric cancer cells (SNU216, N87), and it has lower 50% inhibitory concentration values compared with other EGFR tyrosine kinase inhibitors, including gefitinib, lapatinib, BIBW-2992, and CI-1033. PF299804 induces apoptosis and G1 arrest and inhibits phosphorylation of receptors in the HER family and downstream signaling pathways including STAT3, AKT, and extracellular signal-regulated kinases (ERK) in HER2-amplified gastric cancer cells. PF299804 also blocks EGFR/HER2, HER2/HER3, and HER3/HER4 heterodimer formation as well as the association of HER3 with p85α in SNU216 cells. [3] A recent research uses forty-seven human breast cancer and immortalized breast epithelial lines to evaluate the inhibition effects of PF299804, the results indicate PF299804 preferentially inhibits growth of HER-2-amplified breast cancer cell lines than nonamplified lines (RR = 3.39, p < 0.0001). PF299804 reduces the phosphorylation of HER2, EGFR, HER4, AKT, and ERK in the majority of sensitive lines. PF299804 exerts its anti-proliferative effect through a combined G0/G1 arrest and an induction of apoptosis. [4]
Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
PC9 cells MXjGeY5kfGmxbjDhd5NigQ>? NIm4[W0zKGh? NYTJdXM4UW6qaXLpeIlwdiCxZjDFS2ZTKGW6b36gNVkh\GWuZYTpc44h[WO2aY\heIlv\yCvdYThcpQheGixc4Doc5J6dGG2aX;uJIlvKGi3bXHuJHBEQSClZXzsd{Bi\nSncjCyJIhzeyCkeTDmcJVwemW|Y3XuZ4Uh[XO|YYmsJGlEPTB;MD62N{BvVQ>? NYPaPGdtRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkO5N|A6QTRpPkKzPVMxQTl2PD;hQi=>
human LoVo cells NUDFeWtGTnWwY4Tpc44h[XO|YYm= NUfYSZNJOiCq MXzJcohq[mm2aX;uJI9nKHerbHSgeJlx\SCHR1\SJJBpd3OyaH;yfYxifGmxbjDpckBpfW2jbjDMc3ZwKGOnbHzzJIFnfGW{IEKgbJJ{KGK7IH\seY9z\XOlZX7j[UBie3OjeTygTWM2OD1yLkCxNUDPxE1? NEC4NWQ9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{M{mzNFk6PCd-MkO5N|A6QTR:L3G+
human NCI-H1975 cells Mnn4SpVv[3Srb36gZZN{[Xl? NEnaN5UzKGh? NVrmVVhLUW6qaXLpeIlwdiCxZjDFS2ZTKEx6NUjSM3Q6PzCPIHTveYJt\SCvdYThcpQheGixc4Doc5J6dGG2aX;uJIlvKGi3bXHuJG5EUS2KMUm3OUBk\WyuczDh[pRmeiB{IHjyd{BjgSCobIXvdoV{[2WwY3WgZZN{[XluIFnDOVA:OC5yNEKg{txO NHnpZo49[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{M{mzNFk6PCd-MkO5N|A6QTR:L3G+
human NCI-H1975 cells Mn\RVJJwdGmoZYLheIlwdiCjc4PhfS=> NE\4PGFCdnSrcILvcIln\XKjdHn2[UBi[3Srdnn0fUBi\2GrboP0JIh2dWGwIF7DTU1JOTl5NTDj[YxteyCjc4Pld5Nm\CCjczDndo94fGhiaX7obYJqfGmxbjygS2k2OD1yLkGyN|Mh|ryP M3XhNVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ4M{GwPFkxLz5{NkOxNFg6ODxxYU6=
Sf9 M2TWZWZ2dmO2aX;uJIF{e2G7 MoHjOkBucW6| NILXSYVKenKndnXyd4ljdGViaX7obYJqfGmxbjDv[kBIW1RvdHHn[4VlKEWUQlKxJEh2dmuwb4fuJI9zcWerbjmgLG1mfC14NkigeI8hSWyjLUGyNVEhemW|aXT1[ZMqKGW6cILld5Nm\CCrbjDiZYN2dG:4aYL1d{Bqdm[nY4Tl[EBU\jliaX7z[YN1KGOnbHzzJIF{e2W|c3XkJIF{KHKnZIXjeIlwdiCrbjDHcJUwXHm{IHPvdI9tgW2ncjDwbI9{eGixconsZZRqd25iYX\0[ZIhPiCvaX7zJIJ6KEWOSWPBMEBKSzVyIE2gNE4xODZizszNMi=> NXficm1rRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMke0PVExOjNpPkK3OFkyODJ|PD;hQi=>
NIH/3T3 MVTGeY5kfGmxbjDhd5NigQ>? NUTv[o9ZOiCqcoO= MoT6TZJz\X[ncoPpZoxmKGmwaHnibZRqd25ib3[g[pVtdCCuZX7neIghcHWvYX6gSXJDSjFiYYX0c5Bpd3OyaH;yfYxifGmxbjD0doFve2[nY4Tl[EBqdiCHR1[td5RqdXWuYYTl[EBud3W|ZTDOTWgwO1R|IHPlcIx{KGmwY4XiZZRm\CCob4KgNkBpenNiZn;scI94\WRiYomgd5RqdXWuYYTpc44hf2m2aDDFS2Yh\m:{IEGwJI1qdnNuIFnDOVAhRSByLkCwOkDPxE1w Mn3RQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjd2OUGwNlMoRjJ5NEmxNFI{RC:jPh?=
Sf9 M3XaeWZ2dmO2aX;uJIF{e2G7 MVm2JI1qdnN? MVzJdpJmfmW{c3nicIUhcW6qaXLpeIlwdiCxZjDHV3QufGGpZ3XkJGVTSkJ{IDj1cotvd3ewIH;ybYdqdiliKFns[U03PzVidH:gWoFtNTF{NU[gdoV{cWS3ZYOpJIV5eHKnc4Pl[EBqdiCkYXP1cI93cXK3czDpcoZm[3SnZDDT[lkhcW6|ZXP0JINmdGy|IHHzd4V{e2WmIHHzJJJm\HWldHnvckBqdiCJbIWvWJlzKGOxcH;sfY1meiCyaH;zdIhwenmuYYTpc44h[W[2ZYKgOkBucW6|IHL5JGVNUVODLDDJR|UxKD1iMD6wOFYh|ryPLh?= NIS4W|c9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{N{S5NVAzOyd-Mke0PVExOjN:L3G+
Sf9 NHHacoxHfW6ldHnvckBie3OjeR?= MlHlOkBucW6| NVvGboxrUXK{ZY\ldpNq[mynIHnubIljcXSrb36gc4YhT1OWLYTh[4dm\CCHUlLCOEApfW6tbn;3ckBwemmpaX6pJEhIdHlvMkW5JJRwKEeueT22PVAhemW|aXT1[ZMqKGW6cILld5Nm\CCrbjDiZYN2dG:4aYL1d{Bqdm[nY4Tl[EBU\jliaX7z[YN1KGOnbHzzJIF{e2W|c3XkJIF{KHKnZIXjeIlwdiCrbjDHcJUwXHm{IHPvdI9tgW2ncjDwbI9{eGixconsZZRqd25iYX\0[ZIhPiCvaX7zJIJ6KEWOSWPBMEBKSzVyIE2gNE4xPzRizszNMi=> MWW8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPzR7MUCyN{c,Ojd2OUGwNlM9N2F-
Sf9 MVPGeY5kfGmxbjDhd5NigQ>? MlOyN|AhdWmwcx?= NULkXFBpUXK{ZY\ldpNq[mynIHnubIljcXSrb36gc4YhcHWvYX6gdoVkd22kaX7hcpQhT1OWLYTh[4dm\CCMQVuzJIV5eHKnc4Pl[EBqdiCkYXP1cI93cXK3czDpcoZm[3SnZDDT[lkhcW6|ZXP0JINmdGy|IHHzd4V{e2WmIHHzJJJm\HWldHnvckBqdiCyb3z5[4x2fGGvaXOgZYNq\C22eYLvd4lv\SCyaH;zdIhwenmuYYTpc44h[W[2ZYKgN|AhdWmwczDifUBGVEmVQTygTWM2OCB;IEOuOVch|ryPLh?= MXu8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPzR7MUCyN{c,Ojd2OUGwNlM9N2F-
NIH/3T3 NWK3b5V7TnWwY4Tpc44h[XO|YYm= NIe3N4w{OCCvZz;r[y=> MVmyJIRigXN? M1;NN2lvKH[rdn:gbY5pcWKrdHnvckBw\iCodXzsJIxmdme2aDDoeY1idiCHUlLCNUBifXSxcHjvd5Bpd3K7bHH0bY9vKHS{YX7z[oVkfGWmIHnuJG5KUC9|VEOgZ4VtdHNiaX3wcIFvfGWmIHnuJI1wfXOnIHH0JFMxKG2pL3vnMEBxdyCzZDDmc5IhOiCmYYnzJI1m[XO3cnXkJFI1KGi{czDwc5N1KGyjc4Sg[I9{\SCkeTDX[ZN1\XKwIHLsc5Qh[W6jbInzbZM> NH7CUZI9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{N{S5NVAzOyd-Mke0PVExOjN:L3G+
insect cells NYH2T|I5TnWwY4Tpc44h[XO|YYm= MWDJcohq[mm2aX;uJI9nKEeVVD30ZYdo\WRiaIXtZY4hTUeIUjDjZZRidHm2aXOg[I9u[WmwIHX4dJJme3OnZDDpckBqdnOnY4SgZ4VtdHNuIFnDOVAhRSByLkCwOkDPxE1w MYW8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zQDd3NES3NUc,Ojh5NUS0O|E9N2F-
NCI-H1819 NVHZbW12SW62aYDyc4xq\mW{YYTpeoUh[XO|YYm= M1HmW|czKGi{cx?= NYL4SmdHSW62aYDyc4xq\mW{YYTpeoUh[WO2aY\peJkh[WejaX7zeEBpfW2jbjDOR2kuUDF6MUmgZ4VtdHNiZYjwdoV{e2mwZzD3bYxlKHS7cHWgTGVTOiCrbnP1ZoF1\WRiZn;yJFczKGi{czDifUBOXFNiYYPzZZktKEmFNUCgQUAxNjB{OTFOwG0v NUXBbGo6RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMki3OVQ1PzFpPkK4O|U1PDdzPD;hQi=>
insect cells NF[3V|FHfW6ldHnvckBie3OjeR?= M{fFN2lvcGmkaYTpc44hd2ZiR2PUMZRi\2enZDDoeY1idiCKRWKyJINifGGueYTpZ{Bld22jaX6g[ZhxemW|c3XkJIlvKGmwc3XjeEBk\WyuczygTWM2OCB;IECuNFQ2PyEQvF2u NI\mOlU9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{OEe1OFQ4OSd-Mki3OVQ1PzF:L3G+
NCI-H1975 NVHDToNXSW62aYDyc4xq\mW{YYTpeoUh[XO|YYm= M{fHfVczKGi{cx?= M1\U[2FvfGmycn;sbYZmemG2aY\lJIFkfGm4aYT5JIFo[Wmwc4SgbJVu[W5iTlPJMWgyQTd3IHPlcIx{KGW6cILld5NqdmdiRVfGVkBVPzlyTT;MPFU5WiCvdYThcpQhcW6ldXLheIVlKG[xcjC3NkBpenNiYomgUXRUKGG|c3H5MEBKSzVyIE2gNE41PCEQvF2u M2PJSlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ6N{W0OFcyLz5{OEe1OFQ4OTxxYU6=
Click to View More Cell Line Experimental Data
Assay
Methods Test Index PMID
Western blot pEGFR / EGFR / pERK / ERK / pAKT / AKT / p-mTOR / mTOR / pSTAT3 / STAT3 24853121
Immunofluorescence LC3 28366635
Growth inhibition assay Cell viability 28363995
In vivo Orally administered PF299804 effectively inhibits growth of HCC827 Del/T790M xenografts. [1] Low oral administration of PF-299804 (15mg/kg) causes significant antitumor activity, including marked tumor regressions in a variety of human tumor xenograft models that express and/ or overexpress ERBB family members or contain the double mutation (L858R/T790M) in ERBB1 (EGFR) associated with resistance to gefitinib and erlotinib. [2]

Protocol (from reference)

Kinase Assay:[1]
  • ELISA-Based ERBB Kinase Assay:

    The ERBB1, ERBB 2, and ERBB4 cytoplasmic fusion proteins are made by cloning the ERBB1 sequence (Met-668 to Ala-1211), ERBB2 (Ile-675 to Val-1256), and ERBB4 sequence (Gly-259 to Gly-690) into the baculoviral vector pFastBac using PCR. Proteins are expressed in baculovirusinfected Sf9 insect cells as GST fusion proteins. The proteins are purified by affinity chromatography using glutathione sepharose beads. Inhibition of ERBB tyrosine kinase activity is assessed using an ELISA-based receptor tyrosine kinase assay. Kinase reactions (50 mM HEPES, pH 7.4, 125 mM NaCl, 10 mM MgCl2, 100 μM sodium orthovanadate, 2 mM dithiothreitol, 20 μM ATP, PF299804 or vehicle control, and 1-5 nM GST-erbB per 50 μL of reaction mixture) are run in 96-well plates coated with 0.25 mg/mL poly-Glu-Tyr. The reactions are incubated for 6 minutes at room temperature while being shaken. Kinase reactions are stopped by removal of the reaction mixture, and then the wells are washed with wash buffer (0.1% Tween 20 in PBS). Phosphorylated tyrosine residues are detected by adding 0.2 μg/mL antiphosphotyrosine antibody (Oncogene Ab-4; 50 μL/well) coupled to horseradish peroxidase (HRP) diluted in PBS containing 3% BSA and 0.05% Tween 20 for 25 minutes while being shaken at room temperature. The antibody is removed, and plates are washed in wash buffer. HRP substrate (SureBlue3,3,5,5-tetramethyl benzidine or TMB) is added (50 μL per well) and incubated for 10-20 minutes while it is shaken at room temperature. The TMB reaction is stopped with the addition of 50 μL of stop solution (0.09 N H2SO4). The signal is quantified by measuring absorbance at 450 nm. IC50 values are determined for PF299804 using the median effect method.
Cell Research:[1]
  • Cell lines: Various NSCLC cell lines
  • Concentrations: 0-20 nM
  • Incubation Time: 72 hours
  • Method: Growth and inhibition of growth is assessed by 5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. This assay, a colorimetric method fordetermining the number of viable cells, is based on the bioreduction of MTS by cells to a formazan product that is soluble in cell culture medium, can be detected spectrophotometrically. The cells are exposed totreatment for 72 hours, and the number of cells used per experiment is determined empirically. All experimental points are set up in 6 to 12 wells, and all experiments are repeated at least thrice. The data are graphically displayed using GraphPad Prism version 3.00 for Windows (GraphPad Software). The curves are fitted using a nonlinear regression model with a sigmoidal dose response.
Animal Research:[1]
  • Animal Models: HCC827-GFP or HCC827-Del/T790M lung cancer cells (in 0.2 mL of PBS) are inoculated s.c. into the lower-right quadrant of the flank of nude mice
  • Dosages: 10 mg/kg
  • Administration: Administered via oral gavage

Solubility (25°C)

In vitro

In vivo

Add solvents to the product individually and in order
(Data is from Selleck tests instead of citations):
1% DMSO+30% polyethylene glycol+1% Tween 80, pH 9
For best results, use promptly after mixing.

 10 mg/mL

Chemical Information

Molecular Weight 469.94
Formula

C24H25ClFN5O2
CAS No. 1110813-31-4
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles COC1=C(C=C2C(=C1)N=CN=C2NC3=CC(=C(C=C3)F)Cl)NC(=O)C=CCN4CCCCC4

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

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% 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
NCT04609319 Active not recruiting Drug: Dacomitinib Lung Cancer Pfizer June 11 2021 --
NCT03865446 Completed Drug: Dacomitinib Severe Hepatic Impairment Pfizer April 5 2019 Phase 1
NCT02382796 Completed Drug: Dacomitinib NSCLC Pfizer July 10 2015 Phase 2
NCT02268747 Unknown status Drug: Dacomitinib Skin Squamous Cell Cancer Fondazione IRCCS Istituto Nazionale dei Tumori Milano November 2014 Phase 2
NCT02097433 Completed Drug: Dacomitinib Healthy Pfizer July 2014 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:
I would like to know whether the in vivo formulation you recommend is suitable for oral administration?

Answer:
S2727 in 1% DMSO+30% polyethylene glycol+1% Tween 80 at 10mg/ml is a homogeneous suspension, and it was fine for oral gavage. When preparing the solution, please dissolve the compound in DMSO clearly first. If it dissolves not readily, please sonicate and warm it at about 45-50℃ for a while to help dissolving. Then add PEG 300 and Tween 80. After they mixed well, dilute with water. Then it will become a homogeneous suspension.

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