FAK-IN-9 (Compound 8f) is a potent and orally bioactive FAK inhibitor (antagonist) with IC50 of 27.44 nM. FAK-IN-9 causes apoptosis in triple-negative breast cancer (TNBC) cells.

Physicochemical Properties

Molecular Formula	C36H38CLN7O8S
Molecular Weight	764.25
CAS #	2911655-93-9
PubChem CID	168355641
Appearance	Typically exists as solid at room temperature
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	13
Rotatable Bond Count	20
Heavy Atom Count	53
Complexity	1220
Defined Atom Stereocenter Count	0
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	IC50: 27.44 nM (FAK)[1]
ln Vitro	FAK-IN-9 (Compound 8f; 72 h) suppresses the growth of MDA-MB-157, MDA-MB-231, and MDA-MB-453 cells wIC50 values of 0.167±0.025, 0.126±0.012, and 0.159±0.017 μM, respectively[1]. In a dose-dependent way, MDA-MB-231 cells release relatively significant levels of NO when exposed to FAK-IN-9 (1-4 μM; 72 h)[1]. The invasion and migration of MDA-MB-231 cells is inhibited by FAK-IN-9 (1-4 μM; 48 h) [1]. FAK-IN-9 (1-4 μM; 72 h) efficiently inhibits the signaling pathway controlled by FAK [1]. FAK-IN-9 (4 μM; 72 h) prevents MDA-MB-231 cells from forming stress fibers (SFs) and focal adhesions (FAs) [1]. In MDA-MB-231 cells, FAK-IN-9 (1-4 μM; 72 h) causes apoptosis [1].
ln Vivo	FAK-IN-9 (Compound 8f; oral; once daily for 30 days; 15 or 30 mg/kg) suppresses lung metastases of MDA-MB-231 in mice[1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: MDA-MB-157, MDA-MB-231, MDA-MB-453 and MCF10A Tested Concentrations: Incubation Duration: 72 h Experimental Results: Inhibited proliferation with IC50s of 0.167±0.025, 0.126±0.012, 0.159±0.017 and 2.401±0.131 μM against MDA-MB-157, MDA-MB-231, MDA-MB-453 and MCF10A, respectively. Cell Invasion Assay[1] Cell Types: MDA-MB-231 cells Tested Concentrations: 1, 2 and 4 μM Incubation Duration: 48 h Experimental Results: The numbers of invasive MDA-MB-231 cells were diminished dose-dependently. Cell Migration Assay [1] Cell Types: MDA-MB-231 cells Tested Concentrations: 1, 2 and 4 μM Incubation Duration: 48 h Experimental Results: Remarkably block the migration of MDA-MB-231 cells in a dose-dependent manner. Western Blot Analysis[1] Cell Types: MDA-MB-231 cells Tested Concentrations: 1, 2 and 4 μM Incubation Duration: 72 h Experimental Results: Potently suppressed the autophosphorylation of Y397 in a dose-dependent manner. diminished the levels of p-AKT, MMP-2 and MMP-9 dose dependently. Apoptosis Analysis[1] Cell Types: MDA-MB-231 cells Tested Concentrations: 1, 2 and 4
Animal Protocol	Animal/Disease Models: BALB/c nude mice, MDA-MB-231 experimental pulmonary metastasis model[1] Doses: 15 or 30 mg/kg Route of Administration: po (oral gavage) one time/day for 30 days Experimental Results: Potently decreased the numbers of lung tumor nodules dose- dependently.
References	[1]. Design, synthesis and evaluation of nitric oxide releasing derivatives of 2,4-diaminopyrimidine as novel FAK inhibitors for intervention of metastatic triple-negative breast cancer. Eur J Med Chem. 2023 Mar 15;250:115192.

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	1.3085 mL	6.5424 mL	13.0847 mL
	5 mM	0.2617 mL	1.3085 mL	2.6169 mL
	10 mM	0.1308 mL	0.6542 mL	1.3085 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.