Dacomitinib (PF299804, PF299)

Catalog No.S2727

Molecular Weight(MW): 469.94

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

4 Customer Reviews

J Thorac Oncol, 2018, 13(5):727-731. Dacomitinib (PF299804, PF299) purchased from Selleck.

J Immunol 2014 192(2), 722-31. Dacomitinib (PF299804, PF299) purchased from Selleck.
J Immunol 2014 192(2), 722-31. Dacomitinib (PF299804, PF299) purchased from Selleck.

J Immunol 2014 192(2), 722-31. Dacomitinib (PF299804, PF299) purchased from Selleck.

Purity & Quality Control

Choose Selective EGFR Inhibitors

Biological Activity

Description

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	Incubation Time	Activity Description	PMID
PC9 cells	M3j0XWZ2dmO2aX;uJIF{e2G7	NY\MS2ZKOiCq	NYjDeYdVUW6qaXLpeIlwdiCxZjDFS2ZTKGW6b36gNVkh\GWuZYTpc44h[WO2aY\heIlv\yCvdYThcpQheGixc4Doc5J6dGG2aX;uJIlvKGi3bXHuJHBEQSClZXzsd{Bi\nSncjCyJIhzeyCkeTDmcJVwemW\|Y3XuZ4Uh[XO\|YYmsJGlEPTB;MD62N{BvVQ>?	MYKyN\|k{ODl7NB?=
human LoVo cells	MnnOSpVv[3Srb36gZZN{[Xl?	NYr4NY9MOiCq	M{LKOmlvcGmkaYTpc44hd2Zid3ns[EB1gXCnIFXHSnIheGixc4Doc5J6dGG2aX;uJIlvKGi3bXHuJGxwXm9iY3XscJMh[W[2ZYKgNkBpenNiYomg[ox2d3Knc3PlcoNmKGG\|c3H5MEBKSzVyPUCuNFEyKM7:TR?=	NWi3TJRJOjN7M{C5PVQ>
human NCI-H1975 cells	M1ixRWZ2dmO2aX;uJIF{e2G7	Mke0NkBp	NHvveppKdmirYnn0bY9vKG:oIFXHSnIhVDh3OGKvWFk4OE1iZH;1ZoxmKG23dHHueEBxcG:\|cHjvdplt[XSrb36gbY4hcHWvYX6gUmNKNUhzOUe1JINmdGy\|IHHmeIVzKDJiaILzJIJ6KG[udX;y[ZNk\W6lZTDhd5NigSxiSVO1NF0xNjB2MjFOwG0>	Mo[xNlM6OzB7OUS=
human NCI-H1975 cells	MXzQdo9tcW[ncnH0bY9vKGG\|c3H5		NHLBcWZCdnSrcILvcIln\XKjdHn2[UBi[3Srdnn0fUBi\2GrboP0JIh2dWGwIF7DTU1JOTl5NTDj[YxteyCjc4Pld5Nm\CCjczDndo94fGhiaX7obYJqfGmxbjygS2k2OD1yLkGyN\|Mh\|ryP	NFjv[pczPjNzMEi5NC=>

... Click to View More Cell Line Experimental Data

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

Kinase Assay:^[1]	+ Expand 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]	+ Expand 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. (Only for Reference)
Animal Research:^[1]	+ Expand 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 Formulation: PF299804 is dissolved in DMSO in 10 mM at -20 ° Dosages: 10 mg/kg Administration: Administered via oral gavage (Only for Reference)

References

[4] Kalous O, et al, Mol Cancer Ther, 2012, Jul 3.

+ Expand

Solubility (25°C)

In vitro	DMSO	19 mg/mL (40.43 mM)
	Water	Insoluble
	Ethanol	Insoluble
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

* 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 Download Dacomitinib (PF299804, PF299) SDF

Molecular Weight	469.94
Formula	C₂₄H₂₅ClFN₅O₂
CAS No.	1110813-31-4
Storage	3 years -20°C powder
Storage	2 years -80°C in solvent
Synonyms	N/A

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Clinical Trial Information (data from https://clinicaltrials.gov, updated on 2018-11-16)

NCT Number	Recruitment	Conditions	Sponsor/Collaborators	Start Date	Phases
NCT02382796	Active not recruiting	NSCLC	Pfizer	July 2015	Phase 2
NCT02268747	Unknown status	Skin Squamous Cell Cancer	Fondazione IRCCS Istituto Nazionale dei Tumori Milano	November 2014	Phase 2
NCT02097433	Completed	Healthy	Pfizer	July 2014	Phase 1
NCT02047747	Terminated	Brain Cancer	David Piccioni M.D. Ph.D\|Pfizer\|University of California San Diego	February 2014	Phase 2
NCT02039336	Recruiting	Colorectal Cancer	The Netherlands Cancer Institute\|Pfizer	January 2014	Phase 1\|Phase 2
NCT01920061	Active not recruiting	Neoplasm	Pfizer	September 10 2013	Phase 1

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

Tel: +1-832-582-8158 Ext:3

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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.

EGFR Signaling Pathway Map

EGFR Inhibitors with Unique Features

Selective EGFR Inhibitor

Erlotinib HCl (OSI-744) : EGFR-selective, IC50=2 nM.
Most Potent EGFR Inhibitor

Afatinib (BIBW2992) : EGFR(wt), IC50=0.5 nM; EGFR(L858R), IC50=0.4 nM; EGFR(L858R/T790M), IC50=10 nM; HER2, IC50=14 nM.
FDA-approved EGFR Inhibitor

Lapatinib : Approved by FDA for breast cancer.
Newest EGFR Inhibitor

CO-1686 (AVL-301) ^New : Irreversible, mutant-selective EGFR inhibitor with K_i of 21.5 nM and 303.3 nM for EGFRL858R/T790M and EGFRWT, respectively.

Related EGFR Products4

Erlotinib ^New

Erlotinib is an EGFR inhibitor with IC50 of 2 nM, >1000-fold more sensitive for EGFR than human c-Src or v-Abl.
Afatinib (BIBW2992) Dimaleate ^New

Afatinib (BIBW2992) Dimaleate irreversibly inhibits EGFR/HER2 including EGFR(wt), EGFR(L858R), EGFR(L858R/T790M) and HER2 with IC50 of 0.5 nM, 0.4 nM, 10 nM and 14 nM, respectively; 100-fold more active against Gefitinib-resistant L858R-T790M EGFR mutant.
Poziotinib (HM781-36B) ^New

Poziotinib (HM781-36B) is an irreversible pan-HER inhibitor with IC50 of 3.2 nM, 5.3 nM and 23.5 nM for HER1, HER2, and HER4, respectively. Phase 2.
Gefitinib (ZD1839)

Gefitinib (ZD-1839) is an EGFR inhibitor for Tyr1173, Tyr992, Tyr1173 and Tyr992 in the NR6wtEGFR and NR6W cells with IC50 of 37 nM, 37nM, 26 nM and 57 nM, respectively.

Features:A potent EGFR tyrosine kinase inhibitor.
Erlotinib HCl (OSI-744)

Erlotinib HCl (OSI-744) is an EGFR inhibitor with IC50 of 2 nM in cell-free assays, >1000-fold more sensitive for EGFR than human c-Src or v-Abl.
Afatinib (BIBW2992)

Afatinib (BIBW2992) inhibits EGFR/ErbB irreversibly in vitro with IC50 of 0.5, 0.4, 10, 14, 1 nM for EGFR^wt, EGFR^L858R, EGFR^L858R/T790M ErbB2 (HER2) and ErbB4 (HER4), respectively.
Osimertinib (AZD9291)

Osimertinib (AZD9291) is an oral, irreversible, and mutant-selective EGFR inhibitor with IC50 of 12.92, 11.44 and 493.8 nM for Exon 19 deletion EGFR, L858R/T790M EGFR, and WT EGFR in LoVo cells, respectively. Phase 3.
Lapatinib

Lapatinib, used in the form of Lapatinib Ditosylate, is a potent EGFR and ErbB2 inhibitor with IC50 of 10.8 and 9.2 nM in cell-free assays, respectively.
AG-490 (Tyrphostin B42)

AG-490 (Tyrphostin B42) is an inhibitor of EGFR with IC50 of 0.1 μM in cell-free assays, 135-fold more selective for EGFR versus ErbB2, also inhibits JAK2 with no activity to Lck, Lyn, Btk, Syk and Src.
Rociletinib (CO-1686, AVL-301)

Rociletinib (CO-1686, AVL-301) is an irreversible, mutant-selective EGFR inhibitor with K_i of 21.5 nM and 303.3 nM for EGFR^L858R/T790M and EGFR^WT in cell-free assays, respectively. Phase 2.

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Dacomitinib (PF299804, PF299)