PF-04217903

Validation & Quality Control

Product Citations(1)

Eur J Cancer, 2011, 47(8), 1231-1243

Customer Reviews(1) in vivo invitation

Western blot analysis of c-Met, MAPK and Akt. 0-100nM PF04217903 was added.

Dr. Zhang of Tianjin Medical University. PF-04217903 purchased from Selleck

Quality Control & MSDS

Related Compound Libraries

PF-04217903 is available in the following compound libraries:

Inhibitor Library (96-well) Kinase Inhibitor Library (96-well) Chemical Library (96-well) Tyrosine Kinase Inhibitor Library (96-Well) Cambridge Cancer Compound Library(96-well) Pfizer Licensed Library

Product Information

Compare c-Met Inhibitors

Research Area
Research Area
PF-04217903 Mechanism
PF-04217903 Mechanism
c-Met is an α–β heterodimer with an extracellular α-chain disulfide linked to the membrane-spanning chain harboring the intracellular tyrosine kinase domain. The β-chain consists of the remainder of the Met ectodomain, the transmembrane helix and the cytoplasmic portion. The cytoplasmic portion contains the juxtamembrane domain, kinase domain and a C-terminal tail. ^[1] C-terminal region is essential for downstream signaling and contains two crucial tyrosines (Tyr1349 and Tyr1356). On activation, Tyr1349 and Tyr1356 become auto-phosphorylated and serve as docking sites for a wide spectrum of transducers and adaptors. ^[2]
The crystal structure indicates that PF-04217903 binds to the ATP binding site of non-phosphorylated c-MET kinase domain. PF-04217903 possesses a strong interaction with the activation loop in an auto-inhibitory conformation via a π-π interaction of Tyr1230 with a triazolopyrazine ring, which is characteristic for c-Met. In addition, triazolopyrazine ring is anchored with Tyr1230 and Met1211, and the pyrazole moiety forms hydrogen bonds with a water molecule which interacts with C=O of Tyr1230 and N-H of Arg1086. The ethanolic group on the pyrazole produces the interaction with the activation loop by hydrogen bonds formed by the hydroxyl group to Asp1231 and a water molecule. Besides, a hydrogen bond is also formed between N-3 on the triazolopyrazine ring and the N-H of the DFG Asp1222. Simultaneously, the quinoline group of PF-04217903 is docked into the hinge through a hydrogen bond with Met1160 and hydrophobic interactions with Ile1084, Val1092, Ala1108, and Met1211. The interaction stabilizes the c-MET kinase domain in an unactivated conformation by enhancing the affinity and selectivity of PF-04217903 binding to c-Met. ^[3]
References
[1] Birchmeier C, et al. Nat Rev Mol Cell Biol. 2003, (12), 915-925.
[2] Schiering N, et al. Proc Natl Acad Sci U S A. 2003, 100(22), 12654-12659.
[3] Cui JJ, et al. J Med Chem. 2012, 55(18), 8091-8109.
Dr. Antonino Maria Sparta demonstrates a breakthrough in leukemia research by utilizing two inhibitors produced by Selleck Chemicals.

Cat.No.	Product Name	Solubility (25°C)
S1068	Crizotinib (PF-02341066)	<1 mg/mL	20 mg/mL	<1 mg/mL
S1119	XL-184 free base (Cabozantinib)	<1 mg/mL	100 mg/mL	<1 mg/mL
S1111	Foretinib (GSK1363089, XL880)	<1 mg/mL	127 mg/mL	<1 mg/mL
S1070	PHA-665752	<1 mg/mL	128 mg/mL	<1 mg/mL
S1080	SU11274	<1 mg/mL	92 mg/mL	2 mg/mL
S1094	PF-04217903	<1 mg/mL	5 mg/mL	<1 mg/mL
S1112	SGX-523	<1 mg/mL	2 mg/mL	<1 mg/mL
S1561	BMS 777607	<1 mg/mL	47 mg/mL	<1 mg/mL
S2753	ARQ 197 (Tivantinib)	<1 mg/mL	74 mg/mL	40 mg/mL
S1114	JNJ-38877605	<1 mg/mL	37 mg/mL	<1 mg/mL
S2788	INCB28060	<1 mg/mL	2 mg/mL	<1 mg/mL
S1361	MGCD-265	<1 mg/mL	104 mg/mL	<1 mg/mL
S2774	MK-2461	<1 mg/mL	99 mg/mL	<1 mg/mL
S1316	AMG-208	<<1 mg/mL	0.25 mg/mL	<1 mg/mL
S2859	Golvatinib (E7050)	<1 mg/mL	20 mg/mL	<1 mg/mL
S2747	AMG458	<1 mg/mL	21 mg/mL	<1 mg/mL
S2201	BMS 794833	<1 mg/mL	94 mg/mL	<1 mg/mL
S2761	NVP-BVU972	<1 mg/mL	68 mg/mL	68 mg/mL

Product Description

Biological Activity Protocol Chemical Information Customer Reviews Product Citations Tech Support & FAQs

Biological Activity

Description	PF-04217903 is a selective ATP-competitive c-Met inhibitor with IC50 of 4.8 nM.
Targets	c-Met
IC50	4.8 nM ^[1]
In vitro	Being more selective than staurosporine or PF-02341066, PF-04217903 displays >1000-fold selectivity for c-Met over a panel of 208 kinases, although more susceptible to oncogenic mutations of c-Met that attenuate potency than PF-02341066. In addition to WT c-Met, PF-04217903 displays similar potency to inhibit the activity of c-Met-H1094R, c-Met-R988C, and c-Met-T1010I with IC50 of 3.1 nM, 6.4 nM, and 6.7 nM, respectively, but has no inhibitory activity against c-Met-Y1230C with IC50 of >10 μM. ^[1] PF-04217903 in combination with sunitinib significantly inhibits endothelial cells, but not the tumor cells B16F1, Tib6, EL4, and LLC ^[2] PF-04217903 significantly inhibits the clonogenic growth of LXFA 526L and LXFA 1647L with IC50 values of 16 nM, and 13 nM, respectively, yielding an additive effect when in combination with cetuximab. ^[3] PF-04217903 potently inhibits c-Met-driven processes such as cell growth, motility, invasion, and morphology of a variety of tumor cells. PF-04217903 treatment (2 μM) increased cell death of GTL-16 cells, which involves the downregulation of phosphorylated 4E-BP1, ERK/MAPK associated proteins and PI3K/AKT pathway. ^[4]
In vivo	Although unable to inhibit tumor growth in the sunitinib-sensitive B16F1 and Tib6 tumor models, the combination of PF-04217903 and sunitinib significantly inhibits tumor growth in sunitinib-resistant EL4, and LLC tumor models compared with sunitinib or PF-04217903 alone by significantly blocking vascular expansion, indicating a functional role for HGF/c-Met axis in the sunitinib-resistant tumors. ^[2]
Clinical Trials	A Phase I study to evaluate the effect of PF-04217903 in patients with advanced cancer has been terminated.
Features

Protocol(Only for Reference)

Kinase Assay: ^[1]

Cellular c-Met phosphorylation ELISA	A549 cells with endogenous human WT c-Met are seeded in 96-well plates in growth medium and cultured overnight. On the second day of the assay, the growth medium is replaced with serum-free medium (with 0.04% BSA). Serial dilutions of PF-04217903 are added to each well, and cells are incubated at 37 °C for 1 hour. Then 40 ng/mL HGF is added to the cells for 20 minutes. The cells are washed once with HBSS supplemented with 1 mM Na₃VO₄, and protein lysates are generated from cells using lysis buffer. Phosphorylation of c-Met is assessed by an ELISA method utilizing capture antibodies specific for c-Met and a detection antibody specific for phosphorylated tyrosine residues. Antibody-coated plates are incubated in the presence of protein lysates at 4 °C overnight and washed with 1% Tween 20 in PBS seven times. HRP-PY20 (horseradish peroxidase-conjugated anti-phosphotyrosine) is diluted 1:500 in blocking buffer and added to each plate for 30 minutes. Plates are then washed again, and TMB peroxidase substrate is added to initiate the HRP-dependent colorimetric reaction and the reaction stopped by addition of 0.09 N H₂SO₄. ELISA end points are the absorbance measured at 450 nm using a spectrophotometer. IC50 value is calculated by concentration-response curve fitting utilizing a Microsoft Excel-based four-parameter analytical method.

Cellular c-Met phosphorylation ELISA

A549 cells with endogenous human WT c-Met are seeded in 96-well plates in growth medium and cultured overnight. On the second day of the assay, the growth medium is replaced with serum-free medium (with 0.04% BSA). Serial dilutions of PF-04217903 are added to each well, and cells are incubated at 37 °C for 1 hour. Then 40 ng/mL HGF is added to the cells for 20 minutes. The cells are washed once with HBSS supplemented with 1 mM Na₃VO₄, and protein lysates are generated from cells using lysis buffer. Phosphorylation of c-Met is assessed by an ELISA method utilizing capture antibodies specific for c-Met and a detection antibody specific for phosphorylated tyrosine residues. Antibody-coated plates are incubated in the presence of protein lysates at 4 °C overnight and washed with 1% Tween 20 in PBS seven times. HRP-PY20 (horseradish peroxidase-conjugated anti-phosphotyrosine) is diluted 1:500 in blocking buffer and added to each plate for 30 minutes. Plates are then washed again, and TMB peroxidase substrate is added to initiate the HRP-dependent colorimetric reaction and the reaction stopped by addition of 0.09 N H₂SO₄. ELISA end points are the absorbance measured at 450 nm using a spectrophotometer. IC50 value is calculated by concentration-response curve fitting utilizing a Microsoft Excel-based four-parameter analytical method.

Cell Assay: ^[2]

Cell lines	B16F1, Tib6, EL4, and LLC, HUVECs and C166 cells
Concentrations	Dissolved in DMSO, final concentrations ~2 μM
Incubation Time	4 days
Method	Cells are treated with different concentrations PF-04217903 for 4 days. Cell proliferation is assessed by counting content of each well using a Coulter counter machine.

Animal Study: ^[2]

Animal Models	Immunodeficient nude mice (nu/nu) subcutaneously implanted with tumor cell lines B16F1, EL4, LLC, or Tib6
Formulation	Dissolved in DMSO, and diluted in saline
Dosages	45 mg/kg
Administration	Orally

1

References

Chemical Information

Download PF-04217903 SDF

Molecular Weight (MW)	372.38
Formula	C₁₉H₁₆N₈O
CAS No.	956905-27-4, 956906-93-7 (methanesulfonate), 1159490-81-9 (HCl)
Synonyms	N/A

Solubility (25°C) DMSO 5 mg/mL Water <1 mg/mL Ethanol <1 mg/mL
Storage	2 years -20°CPowder
	2 weeks4°Cin DMSO
	6 months-80°Cin DMSO

Chemical Name	2-(4-(3-(quinolin-6-ylmethyl)-3H-[1,2,3]triazolo[4,5-b]pyrazin-5-yl)-1H-pyrazol-1-yl)ethanol

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Research Area

Customer Reviews (1)

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Rating

Source

Dr. Zhang of Tianjin Medical University. PF-04217903 purchased from Selleck

Method

Western blot

Cell Lines

MDA-MB-231 cells

Concentrations

0-100 nM

Incubation Time

Results

PF-04217903 treatment resulted in a reduction of c-Met, AKT and MAPK phosphorylation in MDA-MB-231 cells.

Product Citations (1)

Primary resistance to cetuximab in a panel of patient-derived tumour xenograft models: activation of MET as one mechanism for drug resistance. [Krumbach R, et al. Eur J Cancer 2011;47(8), 1231-1243]
PubMed: 21273060

Tech Support & FAQs

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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PHA-665752 is a potent, selective and ATP-competitive c-Met inhibitor with IC50 of 9 nM.
SU11274

SU11274 is a selective Met inhibitor with IC50 of 10 nM.
Foretinib (GSK1363089, XL880)

XL880 (GSK1363089, EXEL-2880) is an ATP-competitive inhibitor of MET and KDR with IC50 of 0.4 nM and 0.9 nM, respectively.
SGX-523

SGX-523 is a selective Met inhibitor with IC50 of 4 nM.
JNJ-38877605

JNJ-38877605 is an ATP-competitive inhibitor of c-Met with IC50 of 4 nM.
AMG-208
AMG-208 is a highly selective c-Met inhibitor with IC50 of 9.3 nM.
MGCD-265

MGCD-265 is a potent, multi-target and ATP-competitive inhibitor of c-Met, VEGFR1, VEGFR2, VEGFR3, Ron, and Tie2 with IC50 of 1 nM, 3 nM, 3 nM, 4 nM, 2 nM and 7 nM, respectively.
BMS 777607

BMS-777607 is a Met-related inhibitor for c-Met, Axl, Ron and Tyro3 with IC50 of 3.9 nM, 1.1 nM, 1.8 nM and 4.3 nM, respectively.
BMS 794833
BMS794833 is a potent ATP competitive inhibitor of Met and VEGFR2 with IC50 of 1.7 nM and 15 nM, respectively.
AMG458
AMG 458 is a potent c-Met inhibitor of human c-Met and mouse c-Met with K_i of 1.2 nM or 2.0 nM.

RTKs and Angiogenesis Related Products

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Product Categories

Validation & Quality Control

Product Citations(1)

Customer Reviews(1) in vivo invitation

Quality Control & MSDS

Related Compound Libraries

PF-04217903 is available in the following compound libraries:

Product Information

Compare c-Met Inhibitors

Research Area

PF-04217903 Mechanism

Product Description

Biological Activity

Protocol(Only for Reference)

Kinase Assay: ^[1]

Cell Assay: ^[2]

Animal Study: ^[2]

References

Chemical Information

Research Area

Customer Reviews (1)

Product Citations (1)

Tech Support & FAQs

If you have any other enquiries, please leave a message.

Related c-Met Inhibitors

RTKs and Angiogenesis Related Products

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Product Categories

PF-04217903

Validation & Quality Control

Product Citations(1)

Customer Reviews(1) in vivo invitation

Quality Control & MSDS

Related Compound Libraries

PF-04217903 is available in the following compound libraries:

Product Information

Compare c-Met Inhibitors

Research Area

PF-04217903 Mechanism

Product Description

Biological Activity

Protocol(Only for Reference)

Kinase Assay: [1]

Cell Assay: [2]

Animal Study: [2]

References

Chemical Information

Research Area

Customer Reviews (1)

Product Citations (1)

Tech Support & FAQs

If you have any other enquiries, please leave a message.

Related c-Met Inhibitors

RTKs and Angiogenesis Related Products

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Kinase Assay: ^[1]

Cell Assay: ^[2]

Animal Study: ^[2]