1. JAK/STAT Signaling
    Protein Tyrosine Kinase/RTK
  2. EGFR

EGF816 (Synonyms: Nazartinib)

Cat. No.: HY-12872 Purity: 99.85% ee.: 99.64%
Data Sheet SDS Handling Instructions

EGF816 is a novel, covalent mutant-selective EGFR inhibitor, with Ki and Kinact of 31 nM and 0.222 min−1 on EGFR(L858R/790M) mutant, respectively.

For research use only. We do not sell to patients.
EGF816 Chemical Structure

EGF816 Chemical Structure

CAS No. : 1508250-71-2

Size Price Stock Quantity
10 mM * 1 mL in DMSO $385 In-stock
5 mg $350 In-stock
10 mg $500 In-stock
50 mg $1500 In-stock
100 mg $2100 In-stock
200 mg   Get quote  
500 mg   Get quote  

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Other Forms of EGF816:

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  • Biological Activity

  • Protocol

  • Technical Information

  • Purity & Documentation

  • References

Description

EGF816 is a novel, covalent mutant-selective EGFR inhibitor, with Ki and Kinact of 31 nM and 0.222 min−1 on EGFR(L858R/790M) mutant, respectively.

IC50 & Target

Ki: 31 nM (EGFR(L858R/790M) mutant)[2]

In Vitro

EGF816 has inhibitory effect on the mutant cell lines with IC50s of 4, 6, 2 nM in H1975, H3255, and HCC827, respectively, and demonstrates improved ADME and PK properties[1]. EGF816 shows potent inhibition of pEGFR levels in H3255, HCC827, and H1975 cell lines with EC50 values of 5, 1, and 3 nM, respectively. EGF816 inhibits cell proliferation, with EC50 values of 9, 11, and 25 nM in H3255, HCC827, and H1975, respectively. EGF816 has an OC50 (compound concentration at 50% occupancy) value of 2 and 5 nM on HCC827 and H1975, respectively[2].

In Vivo

In H1975 mouse xenograft model, EGF816 (50 and 20 mg/kg or 25 mg/kg, p.o.) demonstrates dose-dependent efficacy with near complete tumor cells regression at the highest dose tested (50 mg/kg)[1]. In H1975 mouse model, EGF816 (10 mg/kg, p.o.) induces tumor growth inhibition with a T/C (tumor/control volume) of 29%, and when doses are 30 and 100 mg/kg, tumor regressions are achieved (T/C, −61% and −80%, respectively). In the H3255 xenograft model, EGF816 (30 mg/kg, p.o.) shows significant antitumor activity. Antiproliferative activity of EGF816 on 89 lung cancer cell lines indicates that EGF816 selectively inhibits cell lines containing EGFR with catalytic domain mutations[2].

Clinical Trial
NCT Number Sponsor Condition Start Date Phase
NCT02323126 Novartis Pharmaceuticals|Novartis Non Small Cell Lung Cancer February 9, 2015 Phase 2
NCT02108964 Novartis Pharmaceuticals|Novartis Advanced Non-small Cell Lung Cancer June 6, 2014 Phase 1|Phase 2
NCT02335944 Novartis Pharmaceuticals|Novartis Non Small Cell Lung Cancer January 13, 2015 Phase 1|Phase 2
NCT02900664 Novartis Pharmaceuticals|Novartis Colorectal Cancer, Triple Negative Breast Cancer, NSCLC - Adenocarcinoma August 23, 2016 Phase 1
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References
Preparing Stock Solutions
Concentration Volume (DMSO) Mass 1 mg 5 mg 10 mg
1 mM 2.0201 mL 10.1006 mL 20.2012 mL
5 mM 0.4040 mL 2.0201 mL 4.0402 mL
10 mM 0.2020 mL 1.0101 mL 2.0201 mL
Kinase Assay
[1]

Recombinant kinase domain of EGFR L858R and T790M-L858R mutants are incubated with EGF816 to confirm covalent modification of EGFR and site of adduction. Recombinant enzyme is incubated at room temperature with a 20-fold molar excess of compound in 40 mM Tris, pH 8, 500 mM NaCl, 1% glycerol, 5 mM TCEP for 1 h. The reaction is quenched by addition of dithiothreitol (DTT, 80-fold excess to compound) and transfer to ice. A third of the reaction (10 μL) is processed for intact MS by adding an equal volume of 6 M Guan HCl, 100 mM Tris, pH 8, 20 mM DTT, 10 mM TCEP and incubating at room temperature for 15 min. Intact MS analysis is performed on an Agilent 6520 QToF mass spectrometer equipped with a dual spray ion source (IS of 4500 V, fragmentor of 250 V, fas temp of 350°C, and skimmer of 75 V). The samples are injected onto a PLRP-S column (2.1 mm × 50 mm), heated to 60°C, and desalted for 2 min at 500 μL/min and 3% B prior to elution with a fast gradient of 3-50% B in 3 min (B, 0.1% formic acid). The data are analyzed in MassHunter for automatic peak selection, integration, and spectral deconvolution with a mass range of 15 000-75 000 Da. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Cell Assay
[2]

EGF816 is dissolved in DMSO.

Cells are seeded 500 cells/well in solid white 384-well plates in maintenance media. Serial diluted compounds are transferred to cells. After 3 days, cell viability is measured by CellTiter-Glo. BaF3 cell viability is measured 2 days after compound treatment using Bright-Glo Luciferase Assay System. Luminescent readout is normalized to 0.1% DMSO-treated cells and empty wells. Five EGFR TKI-resistant cell lines are generated at Massachusetts General Hospital. MGH134, MGH121, MGH141, and MGH157 are derived from patients who developed resistance to erlotinib with acquired T790M mutation. MGH119-R is generated in vitro by treating MGH119 with gefitinib for a prolonged period. The sensitivity of EGF816 on these lines is tested. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Administration
[1]

EGF816 is formulated in 0.5% MC, 0.5% Tween80 suspension formulation.

Foxn1 nude mice bearing the H1975 tumors are randomized and used for efficacy studies. Compounds are formulated in 0.5% MC, 0.5% Tween 80 suspension formulation and administered by oral gavage at a dosing volume of 10 μL/g of the animal body weight. Animals in each group receive one oral dose of either vehicle (n=6) or the different test compounds (n=6 in each dose group). Plasma samples are collected for PK measurements at 30 min, 3 h, 7 h, and 24 h after last dose on day 5. Body weight is monitored daily, and the % change in body weight is calculated as [(BWcurrent-BWinitial)/(BWinitial)] × 100. Data are presented as percent body weight change from the day of treatment initiation. Tumor sizes are assessed three times during the efficacy study for 5 days. Tumor sizes are determined by using caliper measurements. Tumor volumes are calculated with the formula (length×width×width)/2. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

References
Molecular Weight

495.02

Formula

C₂₆H₃₁ClN₆O₂

CAS No.

1508250-71-2

Storage
Powder -20°C 3 years
  4°C 2 years
In solvent -80°C 6 months
  -20°C 1 month
Shipping

Room temperature in continental US; may vary elsewhere

Solvent & Solubility

DMSO: ≥ 242 mg/mL

* "<1 mg/mL" means slightly soluble or insoluble. "≥" means soluble, but saturation unknown.

Purity: 99.85% ee.: 99.64%

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EGF816
Cat. No.:
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