EGFR-IN-8 is a dual (bifunctional) inhibitor of EGFR and c-Met, compound 48. EGFR-IN-8 is expected to be a target for further development of active molecules targeting EGFR TKI-resistant NSCLC.

Physicochemical Properties

Molecular Formula	C32H23CLF3N7O4
Molecular Weight	662.017735719681
Exact Mass	661.15
Elemental Analysis	C, 58.06; H, 3.50; Cl, 5.35; F, 8.61; N, 14.81; O, 9.67
CAS #	2407957-87-1
PubChem CID	139035057
Appearance	Light yellow to yellow solid powder
LogP	4.9
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	10
Rotatable Bond Count	8
Heavy Atom Count	47
Complexity	1120
Defined Atom Stereocenter Count	0
SMILES	ClC1C=C(C=CC=1C1=NOC(C2=CC=CC(=C2)NC(NC2C=CC(=C(C(F)(F)F)C=2)NC(C2CC2)=O)=O)=N1)NC(C1C=NC=CC=1)=O
InChi Key	IIUNSVLRVWGJRM-UHFFFAOYSA-N
InChi Code	InChI=1S/C32H23ClF3N7O4/c33-25-15-22(38-29(45)19-4-2-12-37-16-19)8-10-23(25)27-42-30(47-43-27)18-3-1-5-20(13-18)39-31(46)40-21-9-11-26(24(14-21)32(34,35)36)41-28(44)17-6-7-17/h1-5,8-17H,6-7H2,(H,38,45)(H,41,44)(H2,39,40,46)
Chemical Name	N-(3-chloro-4-(5-(3-(3-(4-(cyclopropanecarboxamido)-3-(trifluoromethyl)phenyl)ureido)phenyl)-1,2,4-oxadiazol-3-yl)phenyl)nicotinamide
Synonyms	EGFR-IN-8; 2407957-87-1; starbld0000884; CHEMBL4443455; EX-A7176; AKOS040733075; AC-36780
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	In A549, PC9, H1975, CL68, and CL97 cells, EGFR-IN-8 (0–20 μM; 24-72 hours) exhibited time- and dose-dependent inhibitory effects at various time intervals; after 72 hours and 72 hours, respectively, the IC50 values ranged from 48 from 0.3 to 0.6μM and 0.2–0.5μM[1]. Whatever the mutational state of these five cell lines, EGFR-IN-8 (0-0.6 μM; 48 hours) suppresses EGFR and c-Met expression [1].
ln Vivo	EGFR-IN-8 (oral gavage; 50 and 150 mg/kg; once daily; 20 days) demonstrated dose-dependent inhibition of H1975 xenograft tumor growth at doses of 50 and 150 mg/kg (29%, respectively and 60%). 1].
Cell Assay	Cell viability assay [1] Cell Types: A549, PC9, H1975, CL68, CL97 Cell Tested Concentrations: 0-20 μM Incubation Duration: 24 hrs (hours), 48 hrs (hours), 72 hrs (hours) Experimental Results: Inhibitory effect on different cells. Western Blot Analysis[1] Cell Types: A549, PC9, H1975, CL68 and CL97 Cell Tested Concentrations: 0-0.6 μM Incubation Duration: 48 hrs (hours) Experimental Results: diminished EGFR and c-Met expression.
Animal Protocol	Animal/Disease Models: Nude mouse H1975 xenograft tumor model [1] Doses: 50 and 150 mg/kg Route of Administration: po (oral gavage); 50 and 150 mg/kg; one time/day; 20 days Experimental Results: Inhibition of H1975 xenograft tumor growth.
References	[1]. 1,2,4-Oxadiazole derivatives targeting EGFR and c-Met degradation in TKI resistant NSCLC. Eur J Med Chem. 2019 Aug 9;182:111607.

Solubility Data

Solubility (In Vitro)

DMSO : ~8.33 mg/mL (~12.58 mM)

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	1.5105 mL	7.5526 mL	15.1053 mL
	5 mM	0.3021 mL	1.5105 mL	3.0211 mL
	10 mM	0.1511 mL	0.7553 mL	1.5105 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.