Nazartinib mesylate (formerly known as EGF816 mesylate and NVS-816 mesylate) is a novel, covalent/irreversible, and mutant-selective EGFR inhibitor with Ki and Kinact of 31 nM and 0.222 min−1 on EGFR(L858R/790M) mutant, respectively. Nazartinib specifically targets EGFR-activating mutations arising de novo and upon resistance acquisition, while sparing wild-type (WT) EGFR. Non-small cell lung cancer patients carrying oncogenic EGFR mutations initially respond to EGFR-targeted therapy, but later elicit minimal response due to dose-limiting toxicities and acquired resistance.Nazartinib potently inhibits the most common EGFR mutations L858R, Ex19del, and T790M in vitro. The cellular activity of EGF816 on EGFR mutants are assessed using three well-characterized cell lines, H3255, HCC827, and H1975, which harbor the L858R, Ex19del, and L858R/T790M mutations, respectively. Nazartinib is currently under clinical evaluation (phase I/II clinical trials) in patients harboring EGFR mutations, including T790M.

Physicochemical Properties

Molecular Formula	C₂₇H₃₅CLN₆O₅S
Molecular Weight	591.12
Exact Mass	590.207
CAS #	1508250-72-3
Related CAS #	Nazartinib;1508250-71-2;Nazartinib S-enantiomer;1508256-20-9
PubChem CID	91809207
Appearance	Typically exists as solid at room temperature
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	6
Heavy Atom Count	40
Complexity	857
Defined Atom Stereocenter Count	1
SMILES	O=C(C1=CC(C)=NC=C1)NC2=NC3=CC=CC(Cl)=C3N2[C@H]4CN(C(/C=C/CN(C)C)=O)CCCC4.CS(=O)(O)=O
InChi Key	BJRYTAMUVASSBY-ZJULCNDBSA-N
InChi Code	InChI=1S/C26H31ClN6O2.CH4O3S/c1-18-16-19(12-13-28-18)25(35)30-26-29-22-10-6-9-21(27)24(22)33(26)20-8-4-5-15-32(17-20)23(34)11-7-14-31(2)3;1-5(2,3)4/h6-7,9-13,16,20H,4-5,8,14-15,17H2,1-3H3,(H,29,30,35);1H3,(H,2,3,4)/b11-7+;/t20-;/m1./s1
Chemical Name	N-[7-chloro-1-[(3R)-1-[(E)-4-(dimethylamino)but-2-enoyl]azepan-3-yl]benzimidazol-2-yl]-2-methylpyridine-4-carboxamide;methanesulfonic acid
Synonyms	EGF816 mesylate; EGF-816 mesylate; EGF 816 mesylate; NVS-816 mesylate; NVS 816; NVS816; Nazartinib mesylate
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro	Nazartinib (EGF816) exhibits enhanced ADME and PK characteristics and exhibits an inhibitory effect on the mutant cell lines H1975, H3255, and HCC827, with IC50s of 4, 6, and 2 nM, respectively[1]. In H3255, HCC827, and H1975 cell lines, napardinib (EGF816) exhibits strong suppression of pEGFR levels with EC50 values of 5, 1, and 3 nM, respectively. On HCC827 and H1975, napartinib's OC50 (compound concentration at 50% occupancy) values are 2 and 5 nM, respectively[2].
ln Vivo	Nazartinib (EGF816; 50 and 20 mg/kg or 25 mg/kg, po) exhibits dose-dependent effectiveness in the H1975 mouse xenograft model, with nearly total tumor cell regression at the highest dose examined (50 mg/kg)[1]. Nazartinib (EGF816; 10 mg/kg, po) in the H1975 mouse model causes tumor growth inhibition with a T/C (tumor/control volume) of 29%. Tumor regressions are attained at dosages of 30 and 100 mg/kg (T/C, −61% and −80%, respectively). Significant anticancer efficacy is demonstrated by naparsetinib (30 mg/kg, po) in the H3255 xenograft model. Nazartinib selectively inhibits cell lines having EGFR with catalytic domain mutations, as evidenced by its antiproliferative efficacy on 89 lung cancer cell lines[2].
Animal Protocol	Dissolved in 5% ethanol, 25% PEG300, and 70% D5W (5% dextrose in water); 50 mg/kg; Oral administration. balb/c mice and male Wistar rats
References	[1]. Lelais G, et al. Discovery of (R,E)-N-(7-Chloro-1-(1-[4-(dimethylamino)but-2-enoyl]azepan-3-yl)-1H-benzo[d]imidazol-2-yl)-2-methylisonicotinamide (EGF816), a Novel, Potent, and WT Sparing Covalent Inhibitor of Oncogenic (L858R, ex19del) and Resistant (T79 [2]. Jia Y, et al. EGF816 Exerts Anticancer Effects in Non-Small Cell Lung Cancer by Irreversibly and Selectively Targeting Primary and Acquired Activating Mutations in the EGF Receptor. Cancer Res. 2016 Mar 15;76(6):1591-602
Additional Infomation	See also: Nazartinib Mesylate (annotation moved to).

Solubility Data

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.CS(=O)(O)=O  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.6917 mL	8.4585 mL	16.9170 mL
	5 mM	0.3383 mL	1.6917 mL	3.3834 mL
	10 mM	0.1692 mL	0.8459 mL	1.6917 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.