Home Protein Tyrosine Kinase HER2 inhibitor - EGFR inhibitor Neratinib For research use only.

Neratinib

Synonyms: HKI-272

Neratinib is a highly selective HER2 and EGFR inhibitor with IC50 of 59 nM and 92 nM in cell-free assays; weakly inhibits KDR and Src, no significant inhibition to Akt, CDK1/2/4, IKK-2, MK-2, PDK1, c-Raf and c-Met. Phase 3.

Neratinib Chemical Structure

Neratinib Chemical Structure

CAS No. 698387-09-6

Purity & Quality Control

Neratinib Related Products

Signaling Pathway

HER2 Signaling Pathways

HER2 Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
BT-474 Growth Inhibition Assay IC50<0.005 μM 24009064
EFM-192A Growth Inhibition Assay IC50<0.005 μM 24009064
HCC1569 Growth Inhibition Assay IC50<0.005 μM 24009064
HCC1954 Growth Inhibition Assay IC50<0.005 μM 24009064
MDA-MB-175 Growth Inhibition Assay IC50<0.005 μM 24009064
MDA-MB-361 Growth Inhibition Assay IC50<0.005 μM 24009064
SK-BR-3 Growth Inhibition Assay IC50<0.005 μM 24009064
UACC-812 Growth Inhibition Assay IC50<0.005 μM 24009064
UACC-893 Growth Inhibition Assay IC50<0.005 μM 24009064
SUM-225 Growth Inhibition Assay IC50=0.01 μM 24009064
SUM-190 Growth Inhibition Assay IC50=0.01 μM 24009064
ZR-75-1 Growth Inhibition Assay IC50=0.03 μM 24009064
HCC70 Growth Inhibition Assay IC50=0.03 μM 24009064
BT-20 Growth Inhibition Assay IC50=0.07 μM 24009064
MDA-MB-453 Growth Inhibition Assay IC50=0.09 μM 24009064
HCC1187 Growth Inhibition Assay IC50=0.10 μM 24009064
EFM-19 Growth Inhibition Assay IC50=0.11 μM 24009064
T-47D Growth Inhibition Assay IC50=0.16 μM 24009064
MDA-MB-134 Growth Inhibition Assay IC50=0.17 μM 24009064
HCC38 Growth Inhibition Assay IC50=0.25 μM 24009064
MDA-MB-435 Growth Inhibition Assay IC50=0.33 μM 24009064
MDA-MB-468 Growth Inhibition Assay IC50=0.33 μM 24009064
CAMA-1 Growth Inhibition Assay IC50=0.37 μM 24009064
MDA-MB-436 Growth Inhibition Assay IC50=0.41 μM 24009064
MCF-7 Growth Inhibition Assay IC50=0.41 μM 24009064
MDA-MB-415 Growth Inhibition Assay IC50=0.42 μM 24009064
HCC1806 Growth Inhibition Assay IC50=0.44 μM 24009064
HCC1395 Growth Inhibition Assay IC50=0.49 μM 24009064
HCC1937 Growth Inhibition Assay IC50=0.50 μM 24009064
HCC1143 Growth Inhibition Assay IC50=0.54 μM 24009064
UACC-732 Growth Inhibition Assay IC50=0.65 μM 24009064
MDA-MB-231 Growth Inhibition Assay IC50=1.00 μM 24009064
MDA-MB-157 Growth Inhibition Assay IC50=1.12 μM 24009064
BT-549 Growth Inhibition Assay IC50=1.14 μM 24009064
KPL-1 Growth Inhibition Assay IC50=1.89 μM 24009064
CAL-51 Growth Inhibition Assay IC50=1.89 μM 24009064
BT474 Growth Inhibition Assay IC50=0.00323 ± 0.00075 μM 23816254
SKBR3 Growth Inhibition Assay IC50=0.0075 ± 0.005 μM 23816254
MDAMB453 Growth Inhibition Assay IC50=1.59 ± 0.179 μM 23816254
KB Growth Inhibition Assay IC50=4.13 ± 0.47 μM 22491935
KBv200 Growth Inhibition Assay IC50=6.03 ± 0.64 μM 22491935
MCF-7 Growth Inhibition Assay IC50=3.30 ± 0.41 μM 22491935
MCF-7/Adr Growth Inhibition Assay IC50= 2.88 ± 0.30 μM 22491935
MCF-7 Growth Inhibition Assay IC50=3.02 ± 0.34 μM 22491935
MCF-7/FLV1000 Growth Inhibition Assay IC50=7.09 ± 0.71 μM 22491935
HL60 Growth Inhibition Assay IC50=2.26 ± 0.23 μM 22491935
HL60/Adr Growth Inhibition Assay IC50=1.42 ± 0.15 μM 22491935
HEK293/pcDNA3.1 Growth Inhibition Assay IC50=5.29 ± 0.53 μM 22491935
HEK293/ABCB1 Growth Inhibition Assay IC50=6.91 ± 0.70 μM 22491935
SKBR Growth Inhibition Assay 0.01-100 nM 3-7 d inhibits cell growth in time and dose dependent manner 21487605
L858R(EGFR) Cell Viability Assay decreases cell viability in time and dose dependent manner 17311002
L858R/T790M(EGFR) Cell Viability Assay decreases cell viability in time and dose dependent manner 17311002
G776insV_G/C Cell Viability Assay decreases cell viability in time and dose dependent manner 17311002
wild-type Cell Viability Assay decreases cell viability in time and dose dependent manner 17311002
A775insYVMA Cell Viability Assay decreases cell viability in time and dose dependent manner 17311002
G776insV_G/L Cell Viability Assay decreases cell viability in time and dose dependent manner 17311002
P780insGSP Cell Viability Assay decreases cell viability in time and dose dependent manner 17311002
NCI-H1781 Growth Inhibition Assay inhibits cell growth in time and dose dependent manner 16818618
HCC827 Growth Inhibition Assay inhibits cell growth in time and dose dependent manner 16818618
H3255 Growth Inhibition Assay inhibits cell growth in time and dose dependent manner 16818618
NCI-H1975 Growth Inhibition Assay inhibits cell growth in time and dose dependent manner 16818618
A549 Growth Inhibition Assay inhibits cell growth in time and dose dependent manner 16818618
3T3 Growth Inhibition Assay IC50=700 ± 78 nM 15173008
3T3/neu Growth Inhibition Assay IC50=3 ± 0.14 nM 15173008
SK-Br-3 Growth Inhibition Assay IC50=2 ± 0.18 nM 15173008
BT 474 Growth Inhibition Assay IC50=2 ± 0.06 nM 15173008
A431 Growth Inhibition Assay IC50=81 ± 9 nM 15173008
MDA-MB-435 Growth Inhibition Assay IC50=960 ± 165 nM 15173008
SW620 Growth Inhibition Assay IC50=690 ± 84 nM 15173008
SKBR3 Function assay Inhibition of human Her2 in SKBR3 cells, EC50 = 0.002 μM. 18077425
BT474 Function assay Inhibition of human Her2 in BT474 cells, EC50 = 0.002 μM. 18077425
A431 Function assay Inhibition of human Her2 in A431 cells, EC50 = 0.081 μM. 18077425
SW620 Function assay Inhibition of human Her2 in SW620 cells, EC50 = 0.69 μM. 18077425
BA/F3 Cytotoxicity assay Cytotoxicity against mouse BA/F3 cells expressing EGFR L858R mutant, IC50 = 0.0035 μM. 19239229
BA/F3 Cytotoxicity assay Cytotoxicity against mouse BA/F3 cells expressing EGFR L858R/T790M mutant, IC50 = 0.18 μM. 19239229
Sf9 Function assay 10 mins Inhibition of human wild type EGFR expressed in Sf9 cells using [gamma32P]-ATP after 10 mins by scintillation counting, IC50 = 0.0025 μM. 24900643
Sf9 Function assay 10 mins Inhibition of human EGFR T790M/L858R mutant expressed in Sf9 cells using [gamma32P]-ATP after 10 mins by scintillation counting, IC50 = 0.066 μM. 24900643
BAF3 Function assay 72 hrs Inhibition of Tel-fused IGF1R (unknown origin) expressed in mouse BAF3 cells assessed as growth inhibition after 72 hrs by CellTiter-Glo assay, GI50 = 0.19 μM. 28282122
BAF3 Function assay 72 hrs Inhibition of Tel-fused INSR (unknown origin) expressed in mouse BAF3 cells assessed as growth inhibition after 72 hrs by CellTiter-Glo assay, GI50 = 0.29 μM. 28282122
BAF3 Growth inhibition assay 72 hrs Growth inhibition of mouse BAF3 cells after 72 hrs by CellTiter-Glo assay, GI50 = 1.9 μM. 28282122
Click to View More Cell Line Experimental Data

Biological Activity

Description Neratinib is a highly selective HER2 and EGFR inhibitor with IC50 of 59 nM and 92 nM in cell-free assays; weakly inhibits KDR and Src, no significant inhibition to Akt, CDK1/2/4, IKK-2, MK-2, PDK1, c-Raf and c-Met. Phase 3.
Targets
HER2 [1]
(Cell-free assay)		 EGFR [1]
(Cell-free assay)		 KDR [1]
(Cell-free assay)		 Src [1]
(Cell-free assay)
59 nM 92 nM 800 nM 1.4 μM
In vitro
In vitro Neratinib weakly inhibits tyrosine kinases KDR and Src with IC50 of 0.8 μM and 1.4 μM, respectively, being 14- and 24-fold less active compared with HER2. Neratinib displays no activity against other serine-threonine kinases such as Akt, cyclin D1/cdk4, cyclin E/cdk2, cyclin B1/cdk1, IKK-2, MK-2, PDK1, c-Raf, and Tpl-2, as well as the tyrosine kinase c-Met. Neratinib selectively inhibits the proliferation of 3T3 cells transfected with the HER2 (3T3/neu), as well as two other HER-2-overexpressing SK-Br-3 and BT474 cells with IC50 values of 2-3 nM, displaying >230-fold potency compared with non-transfected 3T3 cells as well as MDA-MB-435 and SW620 which are EGFR- and HER2-negative. Neratinib also inhibits the proliferation of EGFR-dependent A431 cells with an IC50 of 81 nM. Neratinib reduces HER2 receptor autophosphorylation in BT474 cells with an IC50 of 5 nM, and EGF-dependent phosphorylation of EGFR in A431 cells with IC50 of 3 nM. Blocking of HER-2 by Neratinib results in inhibition of downstream MAPK and Akt pathways with IC50 of 2 nM, more potently than Trastuzumab. Neratinib inhibits the cyclin D1 expression and the phosphorylation of the Rb-susceptibility gene production in BT474 cells with IC50 of 9 nM, leading to G1-S arrest and ultimately decreased cell proliferation. [1]
Kinase Assay Cell-free autophosphorylation assay using time-resolved fluorometry
Neratinib is prepared as 10 mg/mL stocks in DMSO and diluted in 25 mM HEPES (pH 7.5; 0.002 ng/mL-20 μg/mL). Purified recombinant COOH-terminal fragments of HER2 (amino acids 676-1255) or epidermal growth factor receptor (EGFR) (amino acids 645-1186) [diluted in 100 mM HEPES (pH 7.5) and 50% glycerol] is incubated with increasing concentrations of Neratinib in 4 mM HEPES (pH 7.5), 0.4 mM MnCl2, 20 μM sodium vanadate, and 0.2 mM DTT for 15 minutes at room temperature in 96-well ELISA plates. The kinase reaction is initiated by the addition of 40 μM ATP and 20 mM MgCl2 and allowed to proceed for 1 hour at room temperature. Plates are washed, and phosphorylation is detected using Europium-labeled anti-phospho-tyrosine antibodies (15 ng/well). After washing and enhancement steps, signal is detected using a Victor2 fluorescence reader (excitation wavelength 340 nm, emission wavelength 615 nm). The concentration of Neratinib that inhibits receptor phosphorylation by 50% (IC50) is calculated from inhibition curves.
Cell Research Cell lines 3T3, 3T3/neu, A431, BT474, SK-Br-3, MDA-MB-435, and SW480
Concentrations Dissolved in DMSO, final concentrations 0.5 ng/mL-5 μg/mL
Incubation Time 2 or 6 days
Method Cells are exposed to various concentrations of Neratinib for 2, or 6 days. Cell proliferation is determined using sulforhodamine B, a protein binding dye. Briefly, cells are fixed with 10% trichloroacetic acid and washed extensively with water. Cells are then stained with 0.1% sulforhodamine B and washed in 5% acetic acid. Protein-associated dye is solubilized in 10 mM Tris, and absorbance is measured at 450 nM. The concentration of Neratinib that inhibits cell proliferation by 50% (IC50) is determined from inhibition curves.
Experimental Result Images Methods Biomarkers Images PMID
Western blot p-ERBB2 / ERBB2 / p-ERBB3 / ERBB3 / p-EGFR / p-90RSK pHER2 / HER2 / pAKT / AKT / pERK / ERK 30118499
Growth inhibition assay Cell viability 30118499
In Vivo
In vivo Oral administration of Neratinib significantly inhibits the growth of 3T3/neu xenografts, with inhibition of 34%, 53%, 98%, and 98% at dose of 10, 20, 40, and 80 mg/kg/day, respectively. Consistent with the inhibition of HER-2 phosphorylation by 84% within 1 hour of administration at 40 mg/kg/day, Neratinib inhibits the growth of BT474 xenografts by 70-82%, 67%, and 93% at dose of 5, 10, and 40 mg/kg/day, respectively. Neratinib is also effective against SK-OV-3 xenografts with inhibition of 31% and 85% at 5 and 60 mg/kg/day, respectively. Neratinib is less potent against EGFR-dependent A431 xenografts than HER-2-dependent tumors, with 32% and 44% inhibition at 5 and 20 mg/kg/day, respectively. Neratinib displays little activity against MCF-7 and MX-1 xenografts expressing low levels of HER-2 and EGFR, with only 28% inhibition at 80 mg/kg/day, suggesting that Neratinib has selective activity for cells expressing HER-2 or EGFR. [1]
Animal Research Animal Models Female athymic (nude) mice implanted s.c. with 3T3/neu, BT474, MCF-7, or SK-OV-3 cells
Dosages ~80 mg/kg/day
Administration Gavage
NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT05388149 Recruiting
Breast Cancer|HER2-positive Breast Cancer
University Health Network Toronto
 December 6 2022 Phase 2
NCT05252988 Recruiting
Early-stage Breast Cancer|HER2 Positive Breast Cancer|Hormone Receptor Positive
Spanish Breast Cancer Research Group|Puma Biotechnology Inc.|Pierre Fabre Laboratories
 August 31 2022 Phase 2
NCT04856475 Withdrawn
Breast Cancer|Brain Metastases
Jules Bordet Institute
 November 24 2021 Phase 2
NCT04781374 Withdrawn
Metastatic Prostate Adenocarcinoma|Castration-resistant Prostate Cancer|Prostate Cancer|Prostate Cancer Metastatic
Beth Israel Deaconess Medical Center|Puma Biotechnology Inc.|Dana-Farber Cancer Institute
 May 21 2021 Phase 2

Chemical Information & Solubility

Molecular Weight 557.04 Formula

C30H29ClN6O3
CAS No. 698387-09-6 SDF Download Neratinib SDF
Smiles CCOC1=C(C=C2C(=C1)N=CC(=C2NC3=CC(=C(C=C3)OCC4=CC=CC=N4)Cl)C#N)NC(=O)C=CCN(C)C
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 6 mg/mL ( (10.77 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : Insoluble

Ethanol : Insoluble


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