EGFR kinase inhibitor 1 is a potent EGFR inhibitor (antagonist) with IC50 of 37, 1.7, and >300 nM for WT, l885R/T790M, and L858R/T790M/C797S, respectively. EGFR kinase inhibitor 1 causes apoptosis and cell cycle arrest in the G0/G1 phase. EGFR kinase inhibitor 1 inhibits cell metastasis. EGFR kinase inhibitor 1 has antiproliferation and anti-tumor effects.

Physicochemical Properties

Molecular Formula	C30H31N7O2
Molecular Weight	521.612845659256
Exact Mass	521.253
CAS #	2413958-04-8
PubChem CID	165412576
Appearance	Typically exists as solid at room temperature
LogP	3.8
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	9
Heavy Atom Count	39
Complexity	827
Defined Atom Stereocenter Count	0
InChi Key	YIYTWDRDNVPEHZ-UHFFFAOYSA-N
InChi Code	InChI=1S/C30H31N7O2/c1-6-29(38)32-24-16-25(28(39-5)17-27(24)37-15-9-10-20(37)18-35(2)3)34-30-31-14-13-23(33-30)22-19-36(4)26-12-8-7-11-21(22)26/h6-17,19H,1,18H2,2-5H3,(H,32,38)(H,31,33,34)
Chemical Name	N-[2-[2-[(dimethylamino)methyl]pyrrol-1-yl]-4-methoxy-5-[[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino]phenyl]prop-2-enamide
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	EGFR kinase inhibitor 1 (compound 17i) exhibited antiproliferative activity on A549 and H1975 cells over a 72-hour period, with IC50 values of 4.17 and 0.052 µM, respectively [1]. The induction of apoptosis by EGFR kinase inhibitor 1 (0.05, 0.5, 5 µM; 48 hours) is dose-dependent [1]. In the G0/G1 phase, EGFR kinase inhibitor 1 (4, 20, 100 nM; 48 hours) causes cell cycle arrest [1]. H1975 cell motility is inhibited by EGFR kinase inhibitor 1 (0.5 μM; 0, 24, 48 hours) [1].
Cell Assay	Cytotoxicity assay [1] Cell Types: A549, H1975 Cell Tested Concentrations: Incubation Duration: 72 h Experimental Results: It has anti-proliferative activity against A549 and H1975 cells, with IC50 of 4.17 and 0.052 µM respectively. Apoptosis analysis[1] Cell Types: H1975 Cell Tested Concentrations: 0.05, 0.5, 5 µM Incubation Duration: 48 hrs (hours) Experimental Results: Induction of apoptosis in a dose-dependent manner. Apoptosis analysis [1] Cell Types: H1975 Cell Tested Concentrations: 4, 20, 100 nM Incubation Duration: 48 hrs (hours) Experimental Results: Induced cell cycle arrest in G0/G1 phase, the percentage of cells in G0/G1 phase increased from 42.93% to 4 nM It was 60.52% at 20 nM, 70.39% at 20 nM, and 80.03% at 100 nM.
References	[1]. Design, synthesis and biological evaluation of novel N-(3-amino-4-methoxyphenyl)acrylamide derivatives as selective EGFRL858R/T790M kinase inhibitors. Bioorg Chem. 2022 Jan;118:105471.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.9171 mL	9.5857 mL	19.1714 mL
	5 mM	0.3834 mL	1.9171 mL	3.8343 mL
	10 mM	0.1917 mL	0.9586 mL	1.9171 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.