Name: Neratinib (HKI-272)
Brand: Selleck Chemicals
SKU: S2150
Availability: InStock
Rating: 5 (108 reviews)

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Quality Control

Batch: Purity: 99.94%

99.94

Related Targets	EGFR VEGFR PDGFR FGFR c-Met Src MEK CSF-1R FLT3 c-Kit
Other HER2 Products	CP-724714 Sapitinib (AZD8931) Mubritinib (TAK 165) AC480 (BMS-599626) Tyrphostin AG 879 HER2-Inhibitor-1 TAS0728 Phospho-HER3/ErbB3 (Tyr1197) Antibody [F15P3] Phospho-ErbB3/HER3 (Tyr1289) Antibody [P19F5] Disitamab (Anti-ERBB2 / HER2 / CD340)

Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	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

Solubility

In vitro
Batch:

DMSO : 6 mg/mL (10.77 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

DMSO : 7 mg/mL (12.56 mM) Warmed with 50°C water bath; Ultrasonicated;
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
Mass value Mass unit	Concentration value Concentration unit	Volume value Volume unit	Molecular weight (g/mol)

Dilution Calculator Molecular Weight Calculator

In vivo batch selection In vivo Batch:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	30%PEG400 1 0.5%Tween80 2 5%propylene glycol Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	5.000mg/ml (8.98mM)	Taking the 1 mL working solution as an example, add 300 μL of 16.67 mg/ml clarified PEG400 stock solution to 5 μL of Tween80, mix evenly to clarify it; add 50 μL Propylene glycol to the above system, mix evenly to clarify it; then continue to add 645 μL ddH2O Dilute to 1 mL. The mixed solution should be used immediately for optimal results.

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

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Average weight of animals: g

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Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
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Chemical Information, Storage & Stability

Molecular Weight	557.04	Formula	C₃₀H₂₉ClN₆O₃	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	698387-09-6	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
Synonyms	HKI-272			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

Mechanism of Action

Targets/IC₅₀/Ki	HER2 (Cell-free assay) 59 nM EGFR (Cell-free assay) 92 nM KDR (Cell-free assay) 800 nM Src (Cell-free assay) 1.4 μM
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. This compound 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. It 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. This chemical also inhibits the proliferation of EGFR-dependent A431 cells with an IC50 of 81 nM. It 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 this compound results in inhibition of downstream MAPK and Akt pathways with IC50 of 2 nM, more potently than Trastuzumab. It 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.
Kinase Assay	Cell-free autophosphorylation assay using time-resolved fluorometry
Kinase Assay	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 this compound 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 this chemical that inhibits receptor phosphorylation by 50% (IC50) is calculated from inhibition curves.
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, this compound inhibits the growth of BT474 xenografts by 70-82%, 67%, and 93% at dose of 5, 10, and 40 mg/kg/day, respectively. It is also effective against SK-OV-3 xenografts with inhibition of 31% and 85% at 5 and 60 mg/kg/day, respectively. This chemical 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. It 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 it has selective activity for cells expressing HER-2 or EGFR.
References	[1] https://pubmed.ncbi.nlm.nih.gov/15173008/

Applications

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

Clinical Trial Information

(data from https://clinicaltrials.gov, updated on 2024-05-22)

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

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Neratinib (HKI-272) HER2/EGFR Inhibitor

Chemical Structure

Molecular Weight: 557.04