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Reagentes e instrumentos para imunologia, biologia celular e biologia molecular.

Afatinib [850140-72-6]

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Referência HY-10261-100mg

Tamanho : 100mg

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Telefone : +1 850 650 7790

Marca : MedChemExpress

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Telefone : +1 850 650 7790

Description

Afatinib (BIBW 2992) is an orally active, potent and irreversible dual specificity inhibitor of ErbB family (EGFR and HER2), with IC₅₀ values of 0.5 nM, 0.4 nM, 10 nM and 14 nM for EGFR^wt, EGFR^L858R, EGFR^L858R/T790M and HER2, respectively. Afatinib can be used for the research of esophageal squamous cell carcinoma (ESCC), non-small cell lung cancer (NSCLC) and gastric cancer^[1]^[2]^[3]^[4].

IC₅₀ & Target^[1]

EGFR^L858R

0.4 nM (IC₅₀)

EGFR

0.5 nM (IC₅₀)

EGFR^L858R/T790M

10 nM (IC₅₀)

HER2

14 nM (IC₅₀)

HER3

Cellular Effect

Cell Line	Type	Value	Description	References
A-431	EC₅₀	0.2 μM Compound: Afatinib	Antiproliferative activity against human A431 cells expressing wild type EGFR incubated for 96 hrs measured on day 5 by CellTiterGlo assay Antiproliferative activity against human A431 cells expressing wild type EGFR incubated for 96 hrs measured on day 5 by CellTiterGlo assay	[PMID: 28853575]
A-431	EC₅₀	0.535 μM Compound: Afatinib	Antiproliferative activity against human A-431 cells harboring wild type EGFR measured after 72 hrs by CellTiter-Glo assay Antiproliferative activity against human A-431 cells harboring wild type EGFR measured after 72 hrs by CellTiter-Glo assay	[PMID: 35486541]
A-431	EC₅₀	0.6 μM Compound: Afatinib	Antiproliferative activity against human A431 cells harboring wild type EGFR assessed as growth inhibition after 96 hrs by CellTiter-Glo assay Antiproliferative activity against human A431 cells harboring wild type EGFR assessed as growth inhibition after 96 hrs by CellTiter-Glo assay	[PMID: 28603991]
A-431	GI₅₀	0.594 μM Compound: Afatinib	Antiproliferative activity against human A-431 cells harboring wild type EGFR measured after 72 hrs by CellTiter-Glo assay Antiproliferative activity against human A-431 cells harboring wild type EGFR measured after 72 hrs by CellTiter-Glo assay	[PMID: 35486541]
A-431	IC₅₀	0.02 μM Compound: 5, BIBW2992	Antiproliferative activity against human A431 cells assessed as growth inhibition after 72 hrs by SRB assay Antiproliferative activity against human A431 cells assessed as growth inhibition after 72 hrs by SRB assay	[PMID: 24565969]
A-431	IC₅₀	0.023 nM Compound: 6, BIBW2992	Antiproliferative activity against human A431 cells after 72 hrs by SRB assay Antiproliferative activity against human A431 cells after 72 hrs by SRB assay	[PMID: 24183742]
A-431	IC₅₀	0.9 μM Compound: Afatinib	Antiproliferative activity against human A431 cells after 48 hrs by Celltiter-Glo assay Antiproliferative activity against human A431 cells after 48 hrs by Celltiter-Glo assay	[PMID: 25409491]
A-431	IC₅₀	1.4 μM Compound: Afatinib	Antiproliferation activity against human A431 cells assessed as reduction in cell viability incubated incubated for 48 hrs by MTT assay Antiproliferation activity against human A431 cells assessed as reduction in cell viability incubated incubated for 48 hrs by MTT assay	[PMID: 32912431]
A-431	IC₅₀	10.21 nM Compound: Afatinib	Antiproliferative activity against human A431 cells harboring wild-type EGFR after 72 hrs by CCK8 assay Antiproliferative activity against human A431 cells harboring wild-type EGFR after 72 hrs by CCK8 assay	[PMID: 30600209]
A549	EC₅₀	1.49 μM Compound: Afatinib	Antiproliferative activity against human A549 cells harboring KRAS-G12S mutant incubated for 96 hrs measured on day 5 by CellTiterGlo assay Antiproliferative activity against human A549 cells harboring KRAS-G12S mutant incubated for 96 hrs measured on day 5 by CellTiterGlo assay	[PMID: 28853575]
A549	IC₅₀	0.05 μM Compound: BIBW-2992	Cytotoxicity against human A549 cells after 72 hrs by MTT assay Cytotoxicity against human A549 cells after 72 hrs by MTT assay	[PMID: 26906472]
A549	IC₅₀	0.71 μM Compound: Afatinib	Antiproliferative activity against human A549 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay Antiproliferative activity against human A549 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay	[PMID: 32739648]
A549	IC₅₀	1.2 μM Compound: Afatinib	Antiproliferative activity against human A549 cells expressing wild type EGFR assessed as inhibition of cell growth incubated for 72 hrs by MTT assay Antiproliferative activity against human A549 cells expressing wild type EGFR assessed as inhibition of cell growth incubated for 72 hrs by MTT assay	[PMID: 35728507]
A549	IC₅₀	1.24 μM Compound: Afatinib	Antiproliferative activity against human A549 cells overexpressing c-Met and EGFR assessed as reduction in cell viability incubated for 72 hrs by MTT assay Antiproliferative activity against human A549 cells overexpressing c-Met and EGFR assessed as reduction in cell viability incubated for 72 hrs by MTT assay	[PMID: 37984296]
A549	IC₅₀	1.33 μM Compound: Afatinib	Antiproliferative activity against human A549 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay Antiproliferative activity against human A549 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay	[PMID: 32739648]
A549	IC₅₀	1.4 μM Compound: BIBW-2992	Cytotoxicity against human A549 cells assessed as decrease in cell viability after 72 hrs by MTT assay Cytotoxicity against human A549 cells assessed as decrease in cell viability after 72 hrs by MTT assay	[PMID: 28428040]
A549	IC₅₀	1.811 μM Compound: Afatinib	Cytotoxicity against human A549 cells after 48 hrs by MTT assay Cytotoxicity against human A549 cells after 48 hrs by MTT assay	[PMID: 24900830]
A549	IC₅₀	3766 nM Compound: Afatinib	Antiproliferative activity against human A549 cells harboring wild-type EGFR after 72 hrs by EZ-Cytox assay Antiproliferative activity against human A549 cells harboring wild-type EGFR after 72 hrs by EZ-Cytox assay	[PMID: 30554954]
A549	IC₅₀	5 μM Compound: Afatinib	Antiproliferative activity against wild type human A549 cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay Antiproliferative activity against wild type human A549 cells assessed as reduction in cell viability incubated for 72 hrs by MTT assay	[PMID: 31446247]
A549	IC₅₀	5.4 μM Compound: Afatinib	Antiproliferative activity against wild type human A549 cells assessed as reduction in cell viability incubated for 72 hrs by CCK8 assay Antiproliferative activity against wild type human A549 cells assessed as reduction in cell viability incubated for 72 hrs by CCK8 assay	[PMID: 37141440]
A549	IC₅₀	6.32 μM Compound: Afatinib	Antiproliferative activity against human A549 cells assessed as reduction in cell viability after 24 hrs by MTT assay Antiproliferative activity against human A549 cells assessed as reduction in cell viability after 24 hrs by MTT assay	[PMID: 31648877]
A549	IC₅₀	6.34 μM Compound: Afatinib	Antiproliferative activity against human A549 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay Antiproliferative activity against human A549 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay	[PMID: 32739648]
BaF3	EC₅₀	0.097 μM Compound: Afatinib	Antiproliferative activity against mouse Ba/F3 cells harbouring Her2-A775_G776insYVMA mutant measured after 72 hrs by CellTiter-Glo assay Antiproliferative activity against mouse Ba/F3 cells harbouring Her2-A775_G776insYVMA mutant measured after 72 hrs by CellTiter-Glo assay	[PMID: 35486541]
BaF3	GI₅₀	0.015 μM Compound: Afatinib	Antiproliferative activity against mouse Ba/F3 cells harbouring Her2-A775_G776insYVMA mutant measured after 72 hrs by CellTiter-Glo assay Antiproliferative activity against mouse Ba/F3 cells harbouring Her2-A775_G776insYVMA mutant measured after 72 hrs by CellTiter-Glo assay	[PMID: 35486541]
BaF3	IC₅₀	>1 μM Compound: 6	Antiproliferative activity against mouse BA/F3 cells expressing TEL-JAK3 after 72 hrs by cell titer glo assay Antiproliferative activity against mouse BA/F3 cells expressing TEL-JAK3 after 72 hrs by cell titer glo assay	[PMID: 26258521]
BaF3	IC₅₀	1.58 nM Compound: Afatinib	Antiproliferative activity against mouse BAF3 cells harboring FL-EGFR A763-Y764 ins FQEA mutant after 72 hrs by celltiter-Glo luminescence assay Antiproliferative activity against mouse BAF3 cells harboring FL-EGFR A763-Y764 ins FQEA mutant after 72 hrs by celltiter-Glo luminescence assay	[PMID: 36804726]
BaF3	IC₅₀	1.61 nM Compound: Afatinib	Antiproliferative activity against mouse BAF3 cells harboring HER2-P780-Y781 ins GSP mutant after 72 hrs by celltiter-Glo luminescence assay Antiproliferative activity against mouse BAF3 cells harboring HER2-P780-Y781 ins GSP mutant after 72 hrs by celltiter-Glo luminescence assay	[PMID: 36804726]
BaF3	IC₅₀	19.96 nM Compound: Afatinib	Antiproliferative activity against mouse BaF3 cells harboring EGFR D770_N771insSVD by CellTiter-Glo assay Antiproliferative activity against mouse BaF3 cells harboring EGFR D770_N771insSVD by CellTiter-Glo assay	[PMID: 38518121]
BaF3	IC₅₀	3.8 nM Compound: Afatinib	Antiproliferative activity against mouse BaF3 cells harboring wild type EGFR incubated for 3 days by Cell Titer-Glo assay Antiproliferative activity against mouse BaF3 cells harboring wild type EGFR incubated for 3 days by Cell Titer-Glo assay	[PMID: 37669428]
BaF3	IC₅₀	37.8 nM Compound: Afatinib	Antiproliferative activity against mouse BAF3 cells harboring FL-EGFR D770-N771 ins NPG mutant after 72 hrs by celltiter-Glo luminescence assay Antiproliferative activity against mouse BAF3 cells harboring FL-EGFR D770-N771 ins NPG mutant after 72 hrs by celltiter-Glo luminescence assay	[PMID: 36804726]
BaF3	IC₅₀	381.3 nM Compound: Afatinib	Cytotoxicity against mouse BaF3 cells expressing IL-3 assessed as inhibition of cell viability incubated for 72 hrs by CellTiter-Glo assay Cytotoxicity against mouse BaF3 cells expressing IL-3 assessed as inhibition of cell viability incubated for 72 hrs by CellTiter-Glo assay	[PMID: 28287083]
BaF3	IC₅₀	40 nM Compound: Afatinib	Antiproliferative activity against mouse BaF3 cells harboring EGFR-D770-N771insNPG mutant incubated for 3 days by Cell Titer-Glo assay Antiproliferative activity against mouse BaF3 cells harboring EGFR-D770-N771insNPG mutant incubated for 3 days by Cell Titer-Glo assay	[PMID: 37669428]
BaF3	IC₅₀	9.9 nM Compound: Afatinib	Antiproliferative activity against mouse BAF3 cells harboring HER2-A775-G776 ins YVMA mutant after 72 hrs by celltiter-Glo luminescence assay Antiproliferative activity against mouse BAF3 cells harboring HER2-A775-G776 ins YVMA mutant after 72 hrs by celltiter-Glo luminescence assay	[PMID: 36804726]
BXPC-3	EC₅₀	>1000 nM Compound: Afatinib	Antiproliferative activity against human BxPC3 cells after 72 hrs by MTT assay Antiproliferative activity against human BxPC3 cells after 72 hrs by MTT assay	[PMID: 27010810]
BXPC-3	EC₅₀	>1 μM Compound: Afatinib	Antiproliferative activity against human BxPC3 cells after 72 hrs by MTT assay Antiproliferative activity against human BxPC3 cells after 72 hrs by MTT assay	[PMID: 27010810]
Caco-2	IC₅₀	7.7 μM Compound: Afatinib	Antiproliferative activity against human Caco2 cells by MTT assay Antiproliferative activity against human Caco2 cells by MTT assay	[PMID: 30216848]
Calu-3	GI₅₀	2.77 nM Compound: cpd A	Antiproliferative activity against human Calu-3 cells assessed as reduction in cell viability after 72 hrs by CCK-8 assay Antiproliferative activity against human Calu-3 cells assessed as reduction in cell viability after 72 hrs by CCK-8 assay	[PMID: 35364239]
CL97	IC₅₀	0.1 μM Compound: Afatinib	Antiproliferative activity against human CL97 cells harbouring EGFR G719A/T790M mutant assessed as reduction in cell viability incubated for 72 hrs by MTT assay Antiproliferative activity against human CL97 cells harbouring EGFR G719A/T790M mutant assessed as reduction in cell viability incubated for 72 hrs by MTT assay	[PMID: 31446247]
HCC827	CC₅₀	14 nM Compound: Afatinib	Cytotoxicity in human HCC827 cells assessed as reduction in cell viability incubated for 96 hrs by MTS method Cytotoxicity in human HCC827 cells assessed as reduction in cell viability incubated for 96 hrs by MTS method	[PMID: 31560541]
HCC827	EC₅₀	<0.014 μM Compound: Afatinib	Antiproliferative activity against human HCC827 cells harboring EGFR-delE746_A750 mutant incubated for 96 hrs measured on day 5 by CellTiterGlo assay Antiproliferative activity against human HCC827 cells harboring EGFR-delE746_A750 mutant incubated for 96 hrs measured on day 5 by CellTiterGlo assay	[PMID: 28853575]
HCC827	IC₅₀	0.23 nM Compound: Afatinib	Antiproliferative activity against human HCC827 cells by CellTiter-Glo assay Antiproliferative activity against human HCC827 cells by CellTiter-Glo assay	[PMID: 38518121]
HCC827	IC₅₀	0.4376 nM Compound: Afatinib	Antiproliferative activity against human HCC827 cells harboring EGFR L858R mutant after 72 hrs by CCK8 assay Antiproliferative activity against human HCC827 cells harboring EGFR L858R mutant after 72 hrs by CCK8 assay	[PMID: 30600209]
HCC827	IC₅₀	8 nM Compound: 9, BIBW-2992	Antiproliferative activity against human HCC827 cells after 96 hrs by MTS assay Antiproliferative activity against human HCC827 cells after 96 hrs by MTS assay	[PMID: 20550212]
HCT-116	IC₅₀	11.4 μM Compound: Afatinib	Antiproliferative activity against human HCT116 cells by MTT assay Antiproliferative activity against human HCT116 cells by MTT assay	[PMID: 30216848]
HCT-116	IC₅₀	5.3 μg/mL Compound: Afatinib	Cytotoxicity against human HCT-116 cells Cytotoxicity against human HCT-116 cells	[PMID: 34655985]
HeLa	IC₅₀	6.2 μM Compound: Afatinib	Antiproliferative activity against human HeLa cells by MTT assay Antiproliferative activity against human HeLa cells by MTT assay	[PMID: 30216848]
HeLa	IC₅₀	6.39 μM Compound: Afatinib	Antiproliferative activity against human HeLa cells assessed as reduction in cell viability after 24 hrs by MTT assay Antiproliferative activity against human HeLa cells assessed as reduction in cell viability after 24 hrs by MTT assay	[PMID: 31648877]
HeLa	IC₅₀	6.81 μM Compound: BIBW-2992	Cytotoxicity against human HeLa cells after 72 hrs by MTT assay Cytotoxicity against human HeLa cells after 72 hrs by MTT assay	[PMID: 26906472]
HepG2	IC₅₀	1.16 μM Compound: Afatinib	Antiproliferative activity against human HepG2 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay Antiproliferative activity against human HepG2 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay	[PMID: 32739648]
HepG2	IC₅₀	1.33 μM Compound: BIBW-2992	Cytotoxicity against human HepG2 cells assessed as decrease in cell viability after 72 hrs by MTT assay Cytotoxicity against human HepG2 cells assessed as decrease in cell viability after 72 hrs by MTT assay	[PMID: 28428040]
HepG2	IC₅₀	1.4 μM Compound: Afatinib	Antiproliferative activity against human HepG2 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay Antiproliferative activity against human HepG2 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay	[PMID: 32739648]
HepG2	IC₅₀	5.3 μg/mL Compound: Afatinib	Cytotoxicity against human HepG2 cells Cytotoxicity against human HepG2 cells	[PMID: 34655985]
L02	CC₅₀	25.3 μM Compound: Afatinib	Cytotoxicity against human LO2 cells assessed as reduction in cell viability after 24 hrs by MTT assay Cytotoxicity against human LO2 cells assessed as reduction in cell viability after 24 hrs by MTT assay	[PMID: 31648877]
LoVo	IC₅₀	0.012 μM Compound: 3	Inhibition of wild type EGFR phosphorylation in human LoVo cells after 2 hrs by fluorescence assay Inhibition of wild type EGFR phosphorylation in human LoVo cells after 2 hrs by fluorescence assay	[PMID: 23930994]
LoVo	IC₅₀	15 nM Compound: 11; BIBW-2992	Growth inhibition of human LoVo cells harboring wild type EGFR Growth inhibition of human LoVo cells harboring wild type EGFR	[PMID: 32432477]
MCF7	IC₅₀	0.93 μM Compound: Afatinib	Antiproliferative activity against human MCF7 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay Antiproliferative activity against human MCF7 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay	[PMID: 32739648]
MCF7	IC₅₀	1.09 μM Compound: Afatinib	Antiproliferative activity against human MCF7 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay Antiproliferative activity against human MCF7 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay	[PMID: 32739648]
MCF7	IC₅₀	2.63 μM Compound: Afatinib	Antiproliferative activity against human MCF7 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay Antiproliferative activity against human MCF7 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay	[PMID: 32739648]
MCF7	IC₅₀	2.63 μM Compound: BIBW-2992	Cytotoxicity against human MCF7 cells assessed as decrease in cell viability after 72 hrs by MTT assay Cytotoxicity against human MCF7 cells assessed as decrease in cell viability after 72 hrs by MTT assay	[PMID: 28428040]
MCF7	IC₅₀	5.83 μM Compound: BIBW-2992	Cytotoxicity against human MCF7 cells after 72 hrs by MTT assay Cytotoxicity against human MCF7 cells after 72 hrs by MTT assay	[PMID: 26906472]
MCF7	IC₅₀	7.1 μM Compound: Afatinib	Antiproliferative activity against human MCF7 cells by MTT assay Antiproliferative activity against human MCF7 cells by MTT assay	[PMID: 30216848]
MCF7	IC₅₀	7.62 μM Compound: Afatinib	Antiproliferative activity against human MCF7 cells assessed as reduction in cell viability after 24 hrs by MTT assay Antiproliferative activity against human MCF7 cells assessed as reduction in cell viability after 24 hrs by MTT assay	[PMID: 31648877]
MDA-MB-231	EC₅₀	>1000 nM Compound: Afatinib	Antiproliferative activity against human MDA-MB-231 cells after 72 hrs by MTT assay Antiproliferative activity against human MDA-MB-231 cells after 72 hrs by MTT assay	[PMID: 27010810]
MDA-MB-231	EC₅₀	>1 μM Compound: Afatinib	Antiproliferative activity against human MDA-MB-231 cells after 72 hrs by MTT assay Antiproliferative activity against human MDA-MB-231 cells after 72 hrs by MTT assay	[PMID: 27010810]
MDA-MB-231	IC₅₀	7.5 μM Compound: Afatinib	Antiproliferative activity against human MDA231 cells by MTT assay Antiproliferative activity against human MDA231 cells by MTT assay	[PMID: 30216848]
NCI-H1975	CC₅₀	224 nM Compound: Afatinib	Cytotoxicity in human NCI-H1975 cells assessed as reduction in cell viability incubated for 96 hrs by MTS method Cytotoxicity in human NCI-H1975 cells assessed as reduction in cell viability incubated for 96 hrs by MTS method	[PMID: 31560541]
NCI-H1975	EC₅₀	0.6 μM Compound: Afatinib	Antiproliferative activity against human NCI-H1975 cells harboring EGFR-L858R/T790M double mutant incubated for 96 hrs measured on day 5 by CellTiterGlo assay Antiproliferative activity against human NCI-H1975 cells harboring EGFR-L858R/T790M double mutant incubated for 96 hrs measured on day 5 by CellTiterGlo assay	[PMID: 28853575]
NCI-H1975	EC₅₀	0.7 μM Compound: Afatinib	Antiproliferative activity against human NCI-H1975 cells harboring EGFR L858R/T790M double mutant assessed as growth inhibition after 96 hrs by CellTiter-Glo assay Antiproliferative activity against human NCI-H1975 cells harboring EGFR L858R/T790M double mutant assessed as growth inhibition after 96 hrs by CellTiter-Glo assay	[PMID: 28603991]
NCI-H1975	EC₅₀	116 nM Compound: Afatinib	Antiproliferative activity against human NCI-H1975 cells harboring EGFR L858R/T790M double mutant after 72 hrs by MTT assay Antiproliferative activity against human NCI-H1975 cells harboring EGFR L858R/T790M double mutant after 72 hrs by MTT assay	[PMID: 27010810]
NCI-H1975	GI₅₀	107.1 nM Compound: Afatinib	Antiproliferative activity against human NCI-H1975 cells assessed as growth inhibition Antiproliferative activity against human NCI-H1975 cells assessed as growth inhibition	[PMID: 26310890]
NCI-H1975	IC₅₀	0.023 μM Compound: 3	Inhibition of EGFR L858R/T790M double mutant phosphorylation in human NCI-H1975 cells after 2 hrs by fluorescence assay Inhibition of EGFR L858R/T790M double mutant phosphorylation in human NCI-H1975 cells after 2 hrs by fluorescence assay	[PMID: 23930994]
NCI-H1975	IC₅₀	0.148 μM Compound: 6, BIBW2992	Antiproliferative activity against human NCI-H1975 cells expressing EGFR T790M mutant after 72 hrs by SRB assay Antiproliferative activity against human NCI-H1975 cells expressing EGFR T790M mutant after 72 hrs by SRB assay	[PMID: 24183742]
NCI-H1975	IC₅₀	0.15 μM Compound: 5, BIBW2992	Antiproliferative activity against gefitinib-resistant human NCI-H1975 cells assessed as growth inhibition after 72 hrs by SRB assay Antiproliferative activity against gefitinib-resistant human NCI-H1975 cells assessed as growth inhibition after 72 hrs by SRB assay	[PMID: 24565969]
NCI-H1975	IC₅₀	0.37 μM Compound: Afatinib	Antiproliferative activity against human NCI-H1975 cells harboring EGFR T790M mutant assessed as reduction in cell viability measured after 72 hrs by CCK8 assay Antiproliferative activity against human NCI-H1975 cells harboring EGFR T790M mutant assessed as reduction in cell viability measured after 72 hrs by CCK8 assay	[PMID: 35178185]
NCI-H1975	IC₅₀	0.392 μM Compound: Afatinib	Cytotoxicity against human NCI-H1975 cells after 48 hrs by MTT assay Cytotoxicity against human NCI-H1975 cells after 48 hrs by MTT assay	[PMID: 24900830]
NCI-H1975	IC₅₀	0.9 μM Compound: 39	Antiproliferative activity against human NCI-H1975 cells expressing EGFR L858R/T790M double mutant assessed as decrease in cell viability after 48 hrs by WST assay Antiproliferative activity against human NCI-H1975 cells expressing EGFR L858R/T790M double mutant assessed as decrease in cell viability after 48 hrs by WST assay	[PMID: 26882288]
NCI-H1975	IC₅₀	0.9 μM Compound: Afatinib	Antiproliferative activity against human NCI-H1975 cells after 48 hrs by Celltiter-Glo assay Antiproliferative activity against human NCI-H1975 cells after 48 hrs by Celltiter-Glo assay	[PMID: 25409491]
NCI-H1975	IC₅₀	0.9 μM Compound: Afatinib	Antiproliferative activity against human NCI-H1975 cells overexpressing c-Met and EGFR assessed as reduction in cell viability incubated for 72 hrs by MTT assay Antiproliferative activity against human NCI-H1975 cells overexpressing c-Met and EGFR assessed as reduction in cell viability incubated for 72 hrs by MTT assay	[PMID: 37984296]
NCI-H1975	IC₅₀	0.92 μM Compound: Afatinib	Antiproliferation activity against human NCI-H1975 cells expressing EGFR L858R/T790M mutant assessed as reduction in cell viability incubated incubated for 48 hrs by MTT assay Antiproliferation activity against human NCI-H1975 cells expressing EGFR L858R/T790M mutant assessed as reduction in cell viability incubated incubated for 48 hrs by MTT assay	[PMID: 32912431]
NCI-H1975	IC₅₀	1 μM Compound: Afatinib	Antiproliferative activity against human NCI-H1975 cells harbouring EGFR L858R/T790M double mutant assessed as reduction in cell viability incubated for 72 hrs by MTT assay Antiproliferative activity against human NCI-H1975 cells harbouring EGFR L858R/T790M double mutant assessed as reduction in cell viability incubated for 72 hrs by MTT assay	[PMID: 31446247]
NCI-H1975	IC₅₀	1.97 μM Compound: Afatinib	Cytotoxicity against human NCI-H1975 cells assessed as decrease in cell viability after 72 hrs by CellTiter-Glo assay Cytotoxicity against human NCI-H1975 cells assessed as decrease in cell viability after 72 hrs by CellTiter-Glo assay	[PMID: 28274675]
NCI-H1975	IC₅₀	12.68 μM Compound: Afatinib	Antiproliferative activity against human NCI-H1975 cells assessed as cell growth inhibition by MTT assay Antiproliferative activity against human NCI-H1975 cells assessed as cell growth inhibition by MTT assay	[PMID: 32739648]
NCI-H1975	IC₅₀	131 nM Compound: Afatinib	Antiproliferative activity against human NCI-H1975 cells harboring EGFR T790M/L858R double mutant after 72 hrs by EZ-Cytox assay Antiproliferative activity against human NCI-H1975 cells harboring EGFR T790M/L858R double mutant after 72 hrs by EZ-Cytox assay	[PMID: 30554954]
NCI-H1975	IC₅₀	22 nM Compound: 11; BIBW-2992	Growth inhibition of human NCI-H1975 cells harboring EGFR L858R/T790M double mutant Growth inhibition of human NCI-H1975 cells harboring EGFR L858R/T790M double mutant	[PMID: 32432477]
NCI-H1975	IC₅₀	5.55 μM Compound: Afatinib	Antiproliferative activity against human NCI-H1975 cells assessed as reduction in cell viability after 24 hrs by MTT assay Antiproliferative activity against human NCI-H1975 cells assessed as reduction in cell viability after 24 hrs by MTT assay	[PMID: 31648877]
NCI-H1975	IC₅₀	93 nM Compound: Afatinib	Antiproliferative activity against human NCI-H1975 cells expressing EGFR T790M mutant assessed as inhibition of EGFR phosphorylation Antiproliferative activity against human NCI-H1975 cells expressing EGFR T790M mutant assessed as inhibition of EGFR phosphorylation	[PMID: 35728507]
NCI-H1975	IC₅₀	96.07 nM Compound: Afatinib	Antiproliferative activity against human NCI-H1975 cells harboring EGFR T790M mutant after 72 hrs by CCK8 assay Antiproliferative activity against human NCI-H1975 cells harboring EGFR T790M mutant after 72 hrs by CCK8 assay	[PMID: 30600209]
NCI-H1993	IC₅₀	>12.5 μM Compound: Afatinib	Cytotoxicity against human NCI-H1993 cells after 48 hrs by MTT assay Cytotoxicity against human NCI-H1993 cells after 48 hrs by MTT assay	[PMID: 24900830]
NCI-H358	GI₅₀	1 nM Compound: cpd A	Antiproliferative activity against human NCI-H358 cells assessed as reduction in cell viability after 72 hrs by CCK-8 assay Antiproliferative activity against human NCI-H358 cells assessed as reduction in cell viability after 72 hrs by CCK-8 assay	[PMID: 35364239]
NCI-H460	IC₅₀	6.14 μM Compound: Afatinib	Antiproliferative activity against human H460 cells assessed as reduction in cell viability after 24 hrs by MTT assay Antiproliferative activity against human H460 cells assessed as reduction in cell viability after 24 hrs by MTT assay	[PMID: 31648877]
NIH3T3	IC₅₀	3.605 μM Compound: Afatinib	Cytotoxicity against mouse NIH/3T3 cells after 48 hrs by MTT assay Cytotoxicity against mouse NIH/3T3 cells after 48 hrs by MTT assay	[PMID: 24900830]
NSCLC	IC₅₀	0.5 nM Compound: BIBW-2992	Cytotoxicity against human NSCLC cells Cytotoxicity against human NSCLC cells	[PMID: 26906472]
PC-3	IC₅₀	1.13 μM Compound: Afatinib	Antiproliferative activity against human PC-3 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay Antiproliferative activity against human PC-3 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay	[PMID: 32739648]
PC-3	IC₅₀	2.52 μM Compound: Afatinib	Antiproliferative activity against human PC-3 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay Antiproliferative activity against human PC-3 cells assessed as cell growth inhibition incubated for 72 hrs by MTT assay	[PMID: 32739648]
PC-3	IC₅₀	3.96 μM Compound: BIBW-2992	Cytotoxicity against human PC3 cells assessed as decrease in cell viability after 72 hrs by MTT assay Cytotoxicity against human PC3 cells assessed as decrease in cell viability after 72 hrs by MTT assay	[PMID: 28428040]
PC-3	IC₅₀	4.1 μM Compound: BIBW-2992	Cytotoxicity against human PC3 cells after 72 hrs by MTT assay Cytotoxicity against human PC3 cells after 72 hrs by MTT assay	[PMID: 26906472]
PC-9	GI₅₀	0.37 nM Compound: cpd A	Antiproliferative activity against human PC-9 cells expressing EGFR E746/A750 deletion mutant assessed as reduction in cell viability after 72 hrs by CCK-8 assay Antiproliferative activity against human PC-9 cells expressing EGFR E746/A750 deletion mutant assessed as reduction in cell viability after 72 hrs by CCK-8 assay	[PMID: 35364239]
PC-9	IC₅₀	<0.01 μM Compound: Afatinib	Antiproliferative activity against human PC9 cells harbouring EGFR delE746-A750 mutant assessed as reduction in cell viability incubated for 72 hrs by MTT assay Antiproliferative activity against human PC9 cells harbouring EGFR delE746-A750 mutant assessed as reduction in cell viability incubated for 72 hrs by MTT assay	[PMID: 31446247]
PC-9	IC₅₀	0.00057 μM Compound: 3	Inhibition of EGFR exon 19 deletion activating mutant phosphorylation in human PC9 cells after 2 hrs by fluorescence assay Inhibition of EGFR exon 19 deletion activating mutant phosphorylation in human PC9 cells after 2 hrs by fluorescence assay	[PMID: 23930994]
PC-9	IC₅₀	0.1 nM Compound: Afatinib	Antiproliferative activity against human PC9 cells harboring EGFR exon-19 del mutant after 72 hrs by EZ-Cytox assay Antiproliferative activity against human PC9 cells harboring EGFR exon-19 del mutant after 72 hrs by EZ-Cytox assay	[PMID: 30554954]
PC-9	IC₅₀	0.3 nM Compound: Afatinib	Antiproliferative activity against human PC-9 cells Antiproliferative activity against human PC-9 cells	[PMID: 33429247]
PC-9	IC₅₀	0.57 nM Compound: 3	Inhibition of EGFR exon 19 deletion activating mutant phosphorylation in human PC9 cells after 2 hrs by fluorescence assay Inhibition of EGFR exon 19 deletion activating mutant phosphorylation in human PC9 cells after 2 hrs by fluorescence assay	[PMID: 23930994]
PC-9	IC₅₀	0.638 nM Compound: Afatinib	Anticancer activity against human PC-9 cells harboring del19 mutant assessed as inhibition of cell viability incubated for 72 hrs by CellTiter-Glo assay Anticancer activity against human PC-9 cells harboring del19 mutant assessed as inhibition of cell viability incubated for 72 hrs by CellTiter-Glo assay	[PMID: 28287083]
PC-9	IC₅₀	1.08 μM Compound: Afatinib	Antiproliferative activity against human PC-9 cells overexpressing c-Met and EGFR assessed as reduction in cell viability incubated for 72 hrs by MTT assay Antiproliferative activity against human PC-9 cells overexpressing c-Met and EGFR assessed as reduction in cell viability incubated for 72 hrs by MTT assay	[PMID: 37984296]
PC-9	IC₅₀	1130 nM Compound: Afatinib	Anticancer activity against human PC-9 cells harboring C797S/T790M/del19 mutant assessed as inhibition of cell viability incubated for 72 hrs by CellTiter-Glo assay Anticancer activity against human PC-9 cells harboring C797S/T790M/del19 mutant assessed as inhibition of cell viability incubated for 72 hrs by CellTiter-Glo assay	[PMID: 28287083]
PC-9	IC₅₀	5 nM Compound: 2	Cytotoxicity against human PC-9 cells harboring EGFR mutant assessed as inhibition of cell growth Cytotoxicity against human PC-9 cells harboring EGFR mutant assessed as inhibition of cell growth	[PMID: 36410591]
PC-9	IC₅₀	61.94 nM Compound: Afatinib	Anticancer activity against human PC-9 cells harboring T790M/del19 mutant assessed as inhibition of cell viability incubated for 72 hrs by CellTiter-Glo assay Anticancer activity against human PC-9 cells harboring T790M/del19 mutant assessed as inhibition of cell viability incubated for 72 hrs by CellTiter-Glo assay	[PMID: 28287083]
Sf21	IC₅₀	14 nM Compound: Afatinib	Inhibition of human N-terminal GST-tagged EGFR T790M/L858R double mutant (696 to end residues) expressed in baculovirus infected sf21 cells using poly(Glu, Tyr) 4:1 as substrate Inhibition of human N-terminal GST-tagged EGFR T790M/L858R double mutant (696 to end residues) expressed in baculovirus infected sf21 cells using poly(Glu, Tyr) 4:1 as substrate	[PMID: 30554954]
Sf21	IC₅₀	37 nM Compound: Afatinib	Inhibition of human C-terminal His6-tagged ERBB2 (676 to end residues) expressed in baculovirus infected sf21 cells using poly(Glu, Tyr) 4:1 as substrate Inhibition of human C-terminal His6-tagged ERBB2 (676 to end residues) expressed in baculovirus infected sf21 cells using poly(Glu, Tyr) 4:1 as substrate	[PMID: 30554954]
Sf21	IC₅₀	9 nM Compound: Afatinib	Inhibition of human N-terminal GST-tagged EGFR kinase domain (696 to end residues) expressed in baculovirus infected sf21 cells using poly(Glu, Tyr) 4:1 as substrate Inhibition of human N-terminal GST-tagged EGFR kinase domain (696 to end residues) expressed in baculovirus infected sf21 cells using poly(Glu, Tyr) 4:1 as substrate	[PMID: 30554954]
Sf9	IC₅₀	0.5 nM Compound: 39	Inhibition of human wild type GST-tagged EGFR kinase domain expressed in insect Sf9 cells using pEY (4:1)/biotinylated pEY as substrate after 30 mins by ELISA Inhibition of human wild type GST-tagged EGFR kinase domain expressed in insect Sf9 cells using pEY (4:1)/biotinylated pEY as substrate after 30 mins by ELISA	[PMID: 26882288]
Sf9	IC₅₀	15.4 nM Compound: Afatinib	Inhibition of C-terminal His-tagged N-terminal GST-tagged human EGFR (668 to 1210 residues) expressed in baculovirus infected Sf9 cells after 30 mins in presence of gamma-[32p-ATP] binding assay Inhibition of C-terminal His-tagged N-terminal GST-tagged human EGFR (668 to 1210 residues) expressed in baculovirus infected Sf9 cells after 30 mins in presence of gamma-[32p-ATP] binding assay	[PMID: 31648877]
SK-BR-3	IC₅₀	1.2 nM Compound: Afatinib	Antiproliferative activity against HER2-dependant human SKBR3 cells after 72 hrs by EZ-Cytox assay Antiproliferative activity against HER2-dependant human SKBR3 cells after 72 hrs by EZ-Cytox assay	[PMID: 30554954]
SW-620	IC₅₀	2.13 μM Compound: Afatinib	Antiproliferation activity against EGFR-negative human SW620 cells assessed as reduction in cell viability incubated incubated for 48 hrs by MTT assay Antiproliferation activity against EGFR-negative human SW620 cells assessed as reduction in cell viability incubated incubated for 48 hrs by MTT assay	[PMID: 32912431]

In Vitro

Afatinib (100 nM) sufficiently prevents heregulin-stimulated HER3 phosphorylation^[1].
Afatinib (0-10000 nM) effectively inhibits anchorage-independent proliferation of NIH-3T3 cells ectopically expressing EGFR mutants, and inhibits cell proliferation of H1666, H3255, and NCI 1975 cells^[1].
Afatinib (48-72 h)shows growth inhibition in HKESC-1, HKESC-2, SLMT-1 and EC-1 cells^[2].
Afatinib (0-1 μM, 24-48 h) inhibits AKT and MAPK pathways, and inhibits EGFR and AKT phosphorylation in ESCC cell lines^[2].
Afatinib (0-1 μM, 16-48 h) induces G0/G1 cell cycle arrest in HKESC-2 and EC-1^[2].
Afatinib (0-1 μM, 24-48 h) effectively induces apoptotic cell death in HKESC-2 and EC-1^[2].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Cell Proliferation Assay^[1]

Cell Line:	NIH-3T3 cells, H1666, H3255, and NCI 1975 cells
Concentration:	0, 1, 10, 100, 1000, 10000 nM
Incubation Time:
Result:	Effectively inhibited anchorage-independent proliferation of NIH-3T3 cells ectopically expressing EGFR mutants. Showed inhibition of anchorage independent cell proliferation of various lung cancer cell lines (H1666, H3255, and NCI 1975 cells), with IC₅₀ values of 60 nM, 0.7 nM and 99 nM, respectively.

Cell Viability Assay^[2]

Cell Line:	HKESC-1, HKESC-2, SLMT-1 and EC-1 cell lines
Concentration:
Incubation Time:	48 and 72 hours
Result:	Observed over 95% of growth inhibition. The respective IC₅₀ concentrations at 48 hours (HKESC-1=0.078 μM, HKESC-2=0.115 μM, KYSE510=3.182 μM, SLMT-1=4.625 μM and EC-1=1.489 μM) and 72 hours (HKESC-1=0.002 μM, HKESC-2=0.002 μM, KYSE510=1.090 μM, SLMT-1=1.161 μM and EC-1=0.109 μM) were all in lower micro-molar range.

Western Blot Analysis^[2]

Cell Line:	HKESC-2 cells and EC-1 cells
Concentration:	0, 0.01, and 0.1 μM (HKESC-2 cells), 0, 0.1 and 1 μM (EC-1 cells)
Incubation Time:	24 and 48 hours
Result:	Reduced the phosphorylation of EGFR and the endogenous expression level of HER2 receptors in ESCC cells. Suppressed AKT phosphorylation in a dose and time dependent manner. Significantly reduced the phosphorylation level of the downstream effectors of the AKT-mTOR axis especially in HKESC-2 cells. Inhibited the two major downstream pathways of the ErbB/HER axis, namely, AKT and MAPK pathways in ESCC cell lines.

Cell Cycle Analysis^[2]

Cell Line:	HKESC-2 cells and EC-1 cells
Concentration:	0, 0.01, and 0.1 μM (HKESC-2 cells), 0, 0.1 and 1 μM (EC-1 cells)
Incubation Time:	16, 24, and 48 hours
Result:	Induced G0/G1 cell cycle arrest in both tested ESCC cell lines in a time and dose dependent manner. In HKESC-2 cells, the percentage of cells in G0/G1 phase was increased from 38.2% to 68.1% at 0.01 μM of afatinib and to 74.7% at 0.1 μM of afatinib, from 24 hours (82.4% G0/G1 arrest at 0.01 μM and 86.2% at 0.1 μM) to 48 hours (from 74.7% to 88.2% for 0.01 μM and 91.0% for 0.1 μM). In EC-1 cells, the percentage of cells arrested in the G0/G1 phase was increased from 59.1% to 66.6% and 72.2% at 24 and 48 hours respectively.

Apoptosis Analysis^[2]

Cell Line:	HKESC-2 cells and EC-1 cells
Concentration:	0, 0.01, and 0.1 μM (HKESC-2 cells), 0, 0.1 and 1 μM (EC-1 cells)
Incubation Time:	24 and 48 hours
Result:	Effectively induced cell death by triggering apoptotic mechanisms in ESCC cell lines. Showed a stronger expression level of cleaved Poly (ADP-ribose) polymerase (PARP) in these cell lines.

In Vivo

Afatinib (0-20 mg/kg, Orally, daily for 25 days) shows dramatic tumor regression and downregulation of EGFR, HER2, HER3 and AKT phosphorylation^[1].
Afatinib (15 mg/kg, Orally, in a schedule of 5 days on plus 2 days off, for two weeks) strongly inhibits the growth of HKESC-2 tumor^[2].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Model:	Athymic NMRI-nu/nu female mice (21–31 g, five to six-week-old, transgenic murine lung cancer model and xenograft models)^[1]
Dosage:	15 mg/kg, 20 mg/kg
Administration:	Orally, daily for 25 days
Result:	Resulted in dramatic tumor regression with a cumulative treated/control tumor volume ratio (T/C ratio) of 2% in a standard xenograft model of the epidermoid carcinoma cell line A431, and downregulation of EGFR and AKT phosphorylation. Induced regression of large tumors in this HER2-driven model, effectively controlled xenograft tumor formation by the NCIH1975 cell line, expressing EGFR L858R/T790M, with a T/C value of 12% for doses of 20 mg/kg. Induced more than 50% percent tumor reduction after a 4-week treatment period. Downregulated EGFR, HER2 and HER3 phosphorylation.

Animal Model:	Six weeks old female athymic nude mice (nu/nu) (16-20 g)^[2]
Dosage:	15 mg/kg
Administration:	Oral gavage in a schedule of 5 days on plus 2 days off, for two weeks
Result:	Strongly inhibited the growth of HKESC-2 tumor. Average tumor sizes of vehicle and treatment at end point are 348 ± 24 mm³ and 108 ± 36 mm³ respectively.

Essai clinique

Masse moléculaire

485.94

Formule

C₂₄H₂₅ClFN₅O₃

CAS No.

850140-72-6

Appearance

Solid

Color

White to yellow

SMILES

O=C(NC1=C(C=C2C(C(NC3=CC(Cl)=C(C=C3)F)=NC=N2)=C1)O[C@H]4CCOC4)/C=C/CN(C)C

Livraison

Room temperature in continental US; may vary elsewhere.

Stockage

Powder	-20°C	3 years
	4°C	2 years
In solvent	-80°C	2 years
	-20°C	1 year

Solvant et solubilité

In Vitro:

DMSO : 100 mg/mL (205.79 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions

Concentration Solvent Mass	1 mg	5 mg	10 mg

1 mM	2.0579 mL	10.2893 mL	20.5787 mL
5 mM	0.4116 mL	2.0579 mL	4.1157 mL
10 mM	0.2058 mL	1.0289 mL	2.0579 mL

View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.5 mg/mL (5.14 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Protocol 2

Add each solvent one by one: 10% DMSO 90% Corn Oil
Solubility: ≥ 2.5 mg/mL (5.14 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL Corn oil, and mix evenly.
Protocol 3

Add each solvent one by one: 5% DMSO 40% PEG300 5% Tween-80 50% Saline
Solubility: ≥ 2.5 mg/mL (5.14 mM); Clear solution

For the following dissolution methods, please prepare the working solution directly. It is recommended to prepare fresh solutions and use them promptly within a short period of time.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 0.5% Methylcellulose/saline water
Solubility: 5 mg/mL (10.29 mM); Suspended solution; Need ultrasonic

Pureté et documentation

Purity: 99.92%

Fiche technique (289 KB) SDS (557 KB)

COA (254 KB) LCMS (103 KB)

Instruction de manipulation (2659 KB)

Références

[1]. Li D, et al. BIBW2992, an irreversible EGFR/HER2 inhibitor highly effective in preclinical lung cancer models. Oncogene. 2008 Aug 7;27(34):4702-11. [Content Brief]

[2]. Wong CH, et al. Preclinical evaluation of afatinib (BIBW2992) in esophageal squamous cell carcinoma (ESCC). Am J Cancer Res. 2015 Nov 15;5(12):3588-99. [Content Brief]

[3]. Wang XK, et al. Afatinib circumvents multidrug resistance via dually inhibiting ATP binding cassette subfamily G member 2 in vitro and in vivo. Oncotarget. 2014 Dec 15;5(23):11971-85. [Content Brief]

[4]. Yoshioka T, et al. Antitumor activity of pan-HER inhibitors in HER2-positive gastric cancer. Cancer Sci. 2018 Apr;109(4):1166-1176. [Content Brief]

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Afatinib Related Antibodies

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