Nazartinib S-enantiomer(EGF816)

Molecular Weight (MW)	495.02
Formula	C26H31ClN6O2
CAS No.	1508250-71-2 (free); 1508250-72-3 (mesylate); 1508256-20-9 (S-enantiomer)
Storage	-20℃ for 3 years in powder form
-80℃ for 2 years in solvent
Solubility (In vitro)	DMSO: 99 mg/mL (200mM)
Water: <1 mg/mL
Ethanol: 99 mg/mL (200mM)
Solubility (In vivo)	O=C(NC1=NC2=CC=CC(Cl)=C2N1[C@H]3CN(C(/C=C/CN(C)C)=O)CCCC3)C4=CC=NC(C)=C4
Synonyms	EGF816 S-enantiomer; EGF-816 S-enantiomer; EGF 816 S-enantiomer; NVS-816 S-enantiomer; NVS 816; NVS816; Nazartinib S-enantiomer

Molecular Weight (MW)

495.02

Formula

C26H31ClN6O2

CAS No.

1508250-71-2 (free); 1508250-72-3 (mesylate); 1508256-20-9 (S-enantiomer)

Storage

-20℃ for 3 years in powder form

-80℃ for 2 years in solvent

Solubility (In vitro)

DMSO: 99 mg/mL (200mM)

Water: <1 mg/mL

Ethanol: 99 mg/mL (200mM)

Solubility (In vivo)

O=C(NC1=NC2=CC=CC(Cl)=C2N1[C@H]3CN(C(/C=C/CN(C)C)=O)CCCC3)C4=CC=NC(C)=C4

Synonyms

EGF816 S-enantiomer; EGF-816 S-enantiomer; EGF 816 S-enantiomer; NVS-816 S-enantiomer; NVS 816; NVS816; Nazartinib S-enantiomer

In Vitro	In vitro activity: 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. 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. Kinase Assay: 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. Cell Assay: H1975, H3255, HCC827, A431, and HaCaT cells are maintained in RPMI media supplemented with antibiotics and 10% FBS, maintained in a 37°C, 5% CO2 humidified incubator. After an overnight incubation in 384-well plates, serial diluted compounds are transferred to cells and incubated for 3 hours. HaCaT cells are stimulated with 10 ng/mL EGF (50 ng/mL EGF for A431) for 5 minutes. Cells are lysed in 1% Triton X-100 buffer containing protease and phosphatase inhibitors. Lysates are analyzed by sandwich ELISA utilizing goat anti-EGFR capture antibody, anti-phospho-EGFR(Y1173), and anti-rabbit HRP. Signal is measured by chemiluminescent detection.
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). 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.
Animal model	balb/c mice and male Wistar rats
Formulation & Dosage	Dissolved in 5% ethanol, 25% PEG300, and 70% D5W (5% dextrose in water); 50 mg/kg; Oral administration.
References	Cancer Res. 2016 Mar 15;76(6):1591-602; J Med Chem. 2016 Jul 28;59(14):6671-89.

In Vitro

In vitro activity: 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. 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.

Kinase Assay: 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.

Cell Assay: H1975, H3255, HCC827, A431, and HaCaT cells are maintained in RPMI media supplemented with antibiotics and 10% FBS, maintained in a 37°C, 5% CO2 humidified incubator. After an overnight incubation in 384-well plates, serial diluted compounds are transferred to cells and incubated for 3 hours. HaCaT cells are stimulated with 10 ng/mL EGF (50 ng/mL EGF for A431) for 5 minutes. Cells are lysed in 1% Triton X-100 buffer containing protease and phosphatase inhibitors. Lysates are analyzed by sandwich ELISA utilizing goat anti-EGFR capture antibody, anti-phospho-EGFR(Y1173), and anti-rabbit HRP. Signal is measured by chemiluminescent detection.

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). 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.

Animal model

balb/c mice and male Wistar rats

Formulation & Dosage

Dissolved in 5% ethanol, 25% PEG300, and 70% D5W (5% dextrose in water); 50 mg/kg; Oral administration.

References

Cancer Res. 2016 Mar 15;76(6):1591-602; J Med Chem. 2016 Jul 28;59(14):6671-89.

CAS NO:	1508256-20-9
规格:	≥98%

包装	价格(元)
5mg	电议
10mg	电议
25mg	电议
50mg	电议