FGFR-IN-13 (compound III-30) regulates signaling pathways mediated by endogenous FGFR1 (IC50=0.20±0.02 nM) and FGFR4 (IC50=0.40±0.03 nM) by inhibiting the expression of key proteins. FGFR-IN-13 can inhibit the expression of total-PARP and Bcl-2 proteins and promote the expression of Cleaved-PARP and Bax proteins in a dose-dependent manner. FGFR-IN-13 has significant antitumor activity and is orally active.

Physicochemical Properties

Molecular Formula	C23H21N5O3
Molecular Weight	415.44
CAS #	2962941-25-7
Appearance	Typically exists as solid at room temperature
Density	1.27±0.1 g/cm3(Temp: 20 °C; Press: 760 Torr)(predicted)
LogP	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

ln Vitro	FGFR-IN-13 (10 μM, 9 h) can covalently and irreversibly bind to FGFR protein. Its ability to inhibit p-FGFR protein expression can reach the level of AZD4547 (HY-13330) and TAS-120 (HY-100818) in MDA-MB-231 cell lines [1]. FGFR-IN-13 (2.5, 5, 10 μM, 24 h) can induce cell apoptosis in a dose-dependent manner. At the same concentration (5 μM), its ability to induce apoptosis is better than that of AZD4547 (45.4% and 37.3%). It may have a similar mechanism of action to AZD4547, but MDAMB-231 cells are more sensitive to III-30 [1]. FGFR-IN-13 (1.25, 2.5, 5 μM, 12 h) can effectively inhibit the migration of MDA-MB-231 cells in a dose-dependent manner without causing significant cytotoxicity[1]. FGFR-IN-13 (2.5, 5, 10 μM, 12 h) can induce cell apoptosis by generating excessive ROS and reducing MMP[1].
ln Vivo	FGFR-IN-13 (10 and 30 mg/kg, orally once daily for 21 days) inhibited tumor growth in a dose-dependent manner and had a good safety profile. It could effectively inhibit tumor growth in the MDA-MB-231 xenograft tumor mouse model at a safe dose [1].
Cell Assay	Western Blot Analysis[1] Cell Types: MDA-MB-231 Tested Concentrations: 10 μM Incubation Duration: 9 h Experimental Results: Inhibited the autophosphorylation of FGFR protein for a long period of time, and the expression level of p-FGFR protein in cancer cells is still suppressed even after 8 h of discontinuation. Cell Cycle Analysis[1] Cell Types: MDA-MB-231 Tested Concentrations: 2.5 μM, 5 μM and 10 μM Incubation Duration: 24 h Experimental Results: Induced apoptosis in a dose-dependent manner, and the percentage of induced apoptosis reached 56.8 % at 10 μM. Cell Proliferation Assay[1] Cell Types: MDA-MB- 435, MDA-MB-231, KYSE-150, HepG2 and HUVEC Tested Concentrations: Incubation Duration: 72 h Experimental Results: Showed better inhibitory effect on KYSE-150 cells (IC50 = 1.93 μM) compared with 2 positive controls.
Animal Protocol	Animal/Disease Models:MDA-MB-231 xenograft tumor mouse model[1] Doses: 10 and 30 mg/kg Route of Administration: Oral gavage (p.o.) Experimental Results: Did not cause significant weight loss. Achieved tumor growth inhibition (TGI) of 64.21% at high doses (30 mg/kg). Achieved tumor growth inhibition (TGI) of 40.22% at low doses (10 mg/kg).
References	[1]. Discovery of N-(4-((6-(3, 5-Dimethoxyphenyl)-9H-Purine Derivatives as Irreversible Covalent FGFR Inhibitors[J]. European Journal of Medicinal Chemistry, 2024: 116415.

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	2.4071 mL	12.0354 mL	24.0709 mL
	5 mM	0.4814 mL	2.4071 mL	4.8142 mL
	10 mM	0.2407 mL	1.2035 mL	2.4071 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.