Gefitinib

Biological Activity

Gefitinib (ZD-1839, Iressa) is a novel potent EGFR tyrosine kinase inhibitor with an IC50 of 0.40nM.Gefitinib (ZD-1839,Iressa) selectively inhibited EGF-stimulated growth of HUVECs (IC50 = 0.03 - 0.1 µM) compared with FGF- or VEGF-stimulated growth (IC50 = 1 - 3 µM for both).Gefitinib is a competitive inhibitor with respect to ATP (Ki = 2.1 nM) when the peptide substrate concentration was fixed at 2 mM (6-fold higher than the Km) and the ATP concentration was varied.  Gefitinib showed noncompetitive kinetics (Ki = 15.0 nM) when the ATP concentration was 50 µM (6-fold higher than the Km) and the peptide concentration was varied.

Product Citations

• 

  Endocrinology. 2020 May 30;bqaa086.

  RIPK2 Dictates Insulin Responses to Tyrosine Kinase Inhibitors in Obese Male Mice
  Gefitinib purchased from AbMole

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  Clin Cancer Res. 2018 Nov 15;24(22):5658-5672.

  Increased Synthesis of MCL-1 Protein Underlies Initial Survival of EGFR-Mutant Lung Cancer to EGFR Inhibitors and Provides a Novel Drug Target.
  Gefitinib purchased from AbMole

• 

  Clin Cancer Res. 2018 Jan 1;24(1):197-208.

  Epithelial-to-mesenchymal transition antagonizes response to targeted therapies in lung cancer by suppressing BIM
  Gefitinib purchased from AbMole

• 

  Sci Rep. 2017 May 8;7(1):1578.

  Tyrosine kinase inhibitors of Ripk2 attenuate bacterial cell wallmediated lipolysis, inflammation and dysglycemia
  Gefitinib purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Clinical Cancer Research (2018 Nov). Figure 1. Gefitinib (Abmole Bioscience)
	Method	Apoptosis and cell cycle analysis
	Cell Lines	EGFR mutant NSCLC cell lines
	Concentrations	50 nM
	Incubation Time	72 h
	Results	However, re-exposure to gefitinib over an additional 72 hrs period was inadequate to induce robust apoptosis in PC9 cells, and by the third cycle HCC827 cells also lost their ability to undergo apoptosis.

	Source	Clin Cancer Res (2017). Figure 2. Gefitinib (Abmole Bioscience)
	Method	apoptosis quantified
	Cell Lines	HCC4006 and H1975 cells
	Concentrations	1μM
	Incubation Time	72 h
	Results	We found directly inducing EMT by exogenous TWIST activation (Fig. 2) or chronic TGF- β treatment (Sup. Fig. 2) led to depressed levels of BIM resulting in suppression of EGFRi-induced apoptosis and resistance to EGFRi (WZ4002 and gefitinib).

	Source	Clin Cancer Res (2017). Figure 2. Gefitinib (Abmole Bioscience)
	Method	apoptosis quantified
	Cell Lines	HCC4006 and H1975 cells
	Concentrations	1 μM
	Incubation Time	72 h
	Results	We found directly inducing EMT by exogenous TWIST activation (Fig. 2) or chronic TGF- β treatment (Sup. Fig. 2) led to depressed levels of BIM resulting in suppression of EGFRi-induced apoptosis and resistance to EGFRi (WZ4002 and gefitinib).

	Source	Sci Rep (2017). Figure 7. Gefitinib (AbMole Bioscience, Houston, TX)
	Method	TKI gefitinib inhibits RIPK2-mediated dysglycemia in vivo.
	Cell Lines	C57Bl/6J (WT) and RIPK2−/− mice
	Concentrations	100 mg/kg
	Incubation Time	4 days
	Results	Pre-treatment with gefitinib attenuated lower blood glucose at 6 h post-FK565 injection. At the time of the glucose tolerance test (GTT, 24 h after FK565 injection) gefitinib pre-treatment did not alter fasting blood glucose.

	Source	Sci Rep (2017). Figure 6. Gefitinib (AbMole Bioscience, Houston, TX)
	Method	TKI gefitinib inhibits RIPK2-mediated inflammation in vivo.
	Cell Lines	RIPK2−/− mice
	Concentrations	5–200 mg/kg
	Incubation Time	4 days
	Results	Our results show that treatment with gefitinib at doses equal or greater than 50 mg/kg attenuated Nod1 mediated Cxcl1 protein levels in the circulation of mice in a dose-dependent manner.

	Source	Sci Rep (2017). Figure 5. Gefitinib (AbMole Bioscience, Houston, TX)
	Method	C57Bl/6J (WT) and RIPK2−/− mice
	Cell Lines	HEK293 cells
	Concentrations	1 or 5 μM
	Incubation Time
	Results	We assessed FK565- versus LPS-induced NF-κB activity using HEK293 cells stably over-expressing Nod1 or Tlr4. Consistent with cytokine secretion in adipocytes and macrophages, we find that treatment of HEK Nod1 cells with FK565 (10 μg/mL) increases NF-κB activation, which is attenuated in a dose-dependent manner by pre-incubation with gefitinib at 1 and 5 μM.

	Source	Sci Rep (2017). Figure 4. Gefitinib (AbMole Bioscience, Houston, TX)
	Method
	Cell Lines	bacterial cell
	Concentrations	0.2, 1 or 5 μM
	Incubation Time	1 h
	Results	Time course of FK565-stimulated Cxcl1 secretion in BMDMs (B) n = 4–12. Levels of Cxcl1 released from WT (left panel) or RIPK2−/−(right panel) BMDMs after stimulation with the Tlr4 ligand LPS (0.5 μg/mL) for 48 h and pre-incubated for 1 h with 0.2, 1 or 5 μM gefitinib or SB203580 (C) n = 4–12.

	Source	Sci Rep (2017). Figure 3. Gefitinib (AbMole Bioscience, Houston, TX)
	Method	Westen Blot
	Cell Lines	bacterial cell
	Concentrations	5 μM
	Incubation Time	1 h
	Results	Quantitative comparison was conducted between samples from 4 different blots derived from the same experiment and processed in parallel. (n = 8 total (n = 2/condition per blot).

	Source	Sci Rep (2017). Figure 2. Gefitinib (AbMole Bioscience, Houston, TX)
	Method	TKIs inhibit cytokine response
	Cell Lines	bacterial cell
	Concentrations	1 or 5 μM
	Incubation Time	1 h
	Results	Our results show that pre-incubation with 1–5 μM gefitinib or 1–5 μM SB203580, but not 1–5 μM imatinib significantly attenuated FK565-induced Cxcl1 and Il6 release in a dose-dependent manner in adipocytes.

	Source	Sci Rep (2017). Figure 1. Gefitinib (AbMole Bioscience, Houston, TX)
	Method	bacterial cell wall-induced lipolysis
	Cell Lines	Murine 3T3-L1 preadipocytes
	Concentrations	5 μM
	Incubation Time	48 h
	Results	Only 1 or 5 μM gefitinib and 1 or 5 μM SB203580 dose dependently attenuated the increased rate of glycerol release induced by FK565.

Protocol

Cell Experiment
Cell lines	MDA-361, SKBR-3, MDA-453 and BT-474 cells
Preparation method	Monolayer Growth and Anchorage-independent Growth Assays. For monolayer growth, cells were seeded at a density of 3–4 104 cells in 12-well plates. Twenty-four h later, ZD1839 was added to the cells. Fresh medium ZD1839 was replaced on day 3. On day 5, cells were harvested by trypsinization and counted with a Zeiss Coulter Counter (Beckman Coulter, Miami, FL). Colony-forming assays in soft agarose were performed as described previously (30). Tumor cell colonies measuring 50 m were counted after 7 days using an Omnicon 3800 colony counter and Tumor Colony Analysis V2.IIA software (Imaging Products International, Inc.).
Concentrations	0.1–10 µ M
Incubation time	4 days

Animal Experiment
Animal models	BT-474 cells Xenograft in Athymic Mice
Formulation	0.05% Tween 80
Dosages	200 mg/kg/day for 28 days
Administration	oral gavage

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

Chemical Information

References

Method development and validation for the quantification of dasatinib, erlotinib, gefitinib, imatinib, lapatinib, nilotinib, sorafenib and sunitinib in human plasma by liquid chromatography coupled with tandem mass spectrometry.
Lankheet NA et al. Biomed Chromatogr. 2012 Sep 17. PMID: 22987603.

A pilot study for the early assessment of the effects of BMS-754807 plus gefitinib in an H292 tumor model by [(18)F]fluorothymidine-positron emission tomography.
Lee SJ et al. Invest New Drugs. 2012 Sep 18. PMID: 22987020.

Gefitinib-Induced Paronychia: Response to Autologous Platelet-Rich Plasma.
Kwon SH et al. Arch Dermatol. 2012 Sep 17;1-4. PMID: 22986691.

Polymorphisms to predict outcome to the tyrosine kinase inhibitors gefitinib, erlotinib, sorafenib and sunitinib.
Erdem L et al. Curr Top Med Chem. 2012 Sep 11. PMID: 22978339.