EGFR-IN-11 is a fourth-generation EGFR-tyrosine kinase inhibitor (EGFR-TKI) with IC50 of 18 nM against the triple mutation EGFRL858R/T790M/C797S. EGFR-IN-11 significantly inhibits EGFR phosphorylation, causes apoptosis, and blocks the cell cycle at G0/G1.

Physicochemical Properties

Molecular Formula	C29N9O2SH35
Molecular Weight	573.7123
Exact Mass	573.26
Elemental Analysis	C, 60.71; H, 6.15; N, 21.97; O, 5.58; S, 5.59
CAS #	2463200-44-2
PubChem CID	145712397
Appearance	White to yellow solid powder
LogP	3.7
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	10
Rotatable Bond Count	8
Heavy Atom Count	41
Complexity	971
Defined Atom Stereocenter Count	1
SMILES	N1(CCN(C)CC1)C1C=CC(=CC=1)NC1=NC2N([C@@H]3CCN(C3)S(=O)(=O)C3CC3)C(NC3C=CC=CC=3)=NC=2C=N1
InChi Key	RNEHWYIFHKBZAW-XMMPIXPASA-N
InChi Code	InChI=1S/C29H35N9O2S/c1-35-15-17-36(18-16-35)23-9-7-22(8-10-23)31-28-30-19-26-27(34-28)38(29(33-26)32-21-5-3-2-4-6-21)24-13-14-37(20-24)41(39,40)25-11-12-25/h2-10,19,24-25H,11-18,20H2,1H3,(H,32,33)(H,30,31,34)/t24-/m1/s1
Chemical Name	(R)-9-(1-(cyclopropylsulfonyl)pyrrolidin-3-yl)-N2-(4-(4-methylpiperazin-1-yl)phenyl)-N8-phenyl-9H-purine-2,8-diamine
Synonyms	EGFR-IN-11; RXN00442; RXN-00442; RXN 00442;
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

Targets	EGFRL858R/T790M/C797S 18 nM (IC50)
ln Vitro	EGFR-IN-11 (Compound D9; 0.0001-10 µM; 72 hours) exhibits a highly effective inhibitor of HCC827 and H1975 cell lines, with IC50 values of 0.88 nM and 0.20 µM, correspondingly[1]. EGFR-IN-11 (0.01 EGFR-IN-11 1 µM) inhibits the phosphorylation of EGFR in the HCC827, H1975, and A549 cell line in a concentration-dependent manner[1]. Potently inducing apoptosis in HCC827 cells, EGFR-IN-11 (1 µM; 24 h) also causes cell cycle arrests in HCC827 cells[1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: Human lung cancer cell lines HCC827 (EGFRDel E746-A750), H1975 (EGFRL858R/T790M) and A549 (EGFRWT); epidermoid carcinoma cell line A431 (EGFRWT) Tested Concentrations: 0.0001, 0.0003, 0.001, 0.003, 0.01, 0.1, 1, 10 µM Incubation Duration: 72 hrs (hours) Experimental Results: Inhibited HCC827 H1975 A549 cells proliferation with IC50s of 0.88±0.09 nM, 0.20±0.01 μM, 2.91 ±0.61 μM, and >10 μM, respectively. Western Blot Analysis[1] Cell Types: HCC827, H1975 and A549 cells Tested Concentrations: 1.00, 0.10 and 0.01 µM for HCC827 cells; 10.00, 1.00 and 0.10 µM for H1975 and A549 cells Incubation Duration: 8 hrs (hours) Experimental Results: Suppressed EGFR phosphorylation in a concentration-dependent manner. EGFR phosphorylation in the HCC827 cell line was more remarkably suppressed than in the H1975 and A549 cell lines. Apoptosis Analysis[1] Cell Types: HCC827 cells Tested Concentrations: 1 µM Incubation Duration: 24 hrs (hours) Experimental Results: The percentages of apoptotic cells is 56.91%. Cell Cycle Analysis[1] Cell Types: HCC827 cells Concen
References	[1]. Discovery of novel 9-heterocyclyl substituted 9H-purines as L858R/T790M/C797S mutant EGFR tyrosine kinase inhibitors. Eur J Med Chem. 2019 Nov 16:111888.

Solubility Data

Solubility (In Vitro)

DMSO : 125 mg/mL (217.88 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (3.63 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (3.63 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.7430 mL	8.7152 mL	17.4304 mL
	5 mM	0.3486 mL	1.7430 mL	3.4861 mL
	10 mM	0.1743 mL	0.8715 mL	1.7430 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.