Icotinib

 

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Biological Activity

Description	Icotinib is a potent and specific EGFR inhibitor with IC50 of 5 nM, including the EGFR, EGFR(L858R), EGFR(L861Q), EGFR(T790M) and EGFR(T790M, L858R). Phase 4.
Targets	EGFR
IC50	5 nM [1]
In vitro	Icotinib inhibits EGFR activity in a dose-dependent manner, with an IC50 value of 5 nM and complete inhibition at 62.5 nM. Icotinib selectively solely inhibits the EGFR members including the wild type and mutants with inhibition efficacies of 61-99%. Icotinib blocks EGFR-mediated intracellular tyrosine phosphorylation in human epidermoid carcinoma A431 cells in a dose-dependent manner. Meanwhile, in our proliferation assay performed on A431, BGC-823, A549, H460, HCT8, KB and Bel-7402 cell lines, we found that the relative sensitivity of cell lines to Icotinib is A431 > BGC-823 > A549 > H460 > KB > HCT8 and Bel-7402. Icotinib exhibits a broad spectrum of antitumor activity and it is especially effective against tumors expressing higher levels of EGFR. [1]
In vivo	Icotinib shows an antitumor effect in different types of xenografts. Icotinib inhibits tumor growth at a rate of 51.5%, 31.0% and 67.4% in the A431, A549 and H460 xenografts at a dose of 120 mg/kg, respectively. [1]
Features

Protocol(Only for Reference)

Kinase Assay: ^[1]

Biochemical kinase assays

In the in vitro kinase assays, 2.4 ng/μL EGFR protein is mixed with 32 ng/μL Crk in 25 μL kinase reaction buffer containing 1 μM cold ATP and 1 μCi 32P-γ-ATP. The mix is incubated with Icotinib at 0, 0.5, 2.5, 12.5 or 62.5 nM on ice for 10 min followed by incubation at 30 °C for 20 min. After quenching with SDS sample buffer at 100 °C for 4 min, the protein mix is resolved by electrophoresis in a 10% SDS-PAGE gel. The dried gel is then exposed to the PhosphorImager to detect radioactivity. Quantification is performed by ImageQuant software. In this methodology the radioactive signal inversely correlates with kinase activity.

Cell Assay: ^[1]

Cell lines	A431, BGC-823, A549, H460, HCT8, KB and Bel-7402 cells.
Concentrations	~25 μM
Incubation Time	96 h
Method	Cells (103 /well) are seeded into 96-well plates in RPMI-1640 medium containing 10% FBS and grown in a 5% CO2 incubator at 37 °C. After 24 h, cells are treated with Icotinib at 0, 0.78, 1.56, 3.125, 6.25, 12.5 or 25 μM for 96 h. Cell proliferation is calculated by subtracting the mean absorbance value on day 0 from the mean absorbance value on day 4.

Animal Study: ^[1]

Animal Models	A431, A549, H460 and HCT8 xenografts Nude mice
Dosages	30, 60 and 120 mg/kg/day
Administration	p.o.
Solubility	0.5% CMC, 30 mg/mL

1

References

Clinical Trial Information( data from http://clinicaltrials.gov)

NCT Number	Recruitment	Conditions	Sponsor /Collaborators	Start Date	Phases
NCT01690390	Recruiting	NSCLC	Zhejiang Beta Pharma Inc.	2012-09	Phase 2
NCT01707329	Recruiting	Non-small Cell Lung Cancer	Zhejiang Beta Pharma Inc.	2012-09	Phase 2
NCT01744925	Recruiting	Non-small Cell Lung Cancer	Tianjin Medical University Cancer Institute and Hospital	2012-10	Phase 2
NCT01843647	Active, not recruiting	Non-small-cell Lung Cancer	Zhejiang Beta Pharma Inc.	2013-04	Phase 2
NCT01855854	Active, not recruiting	Adenocarcinoma of the Gastroesophageal Junction|Esophageal Carcinoma	Zhejiang Beta Pharma Inc.	2013-05	Phase 2

Chemical Information

Download Icotinib SDF

Chemical Name	[1,4,7,10]Tetraoxacyclododecino[2,3-g]quinazolin-4-amine, N-(3-ethynylphenyl)-7,8,10,11,13,14-hexahydro-

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	0.39142	3.9142	7.8284	11.7426