FLT3-IN-14 is a potent FLT3 inhibitor (antagonist) with IC50 of 5.6 nM and 1.4 nM for FLT3-WT and FLT3-ITD, respectively. FLT3-IN-14 reduces the phosphorylation of FLT3 (Y591), arrests the cell cycle in the G1 phase, and causes apoptosis.

Physicochemical Properties

Molecular Formula	C25H24N6O2S
Molecular Weight	472.562063217163
Exact Mass	472.168
CAS #	2620551-45-1
PubChem CID	163322280
Appearance	Typically exists as solid at room temperature
LogP	5.4
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	4
Heavy Atom Count	34
Complexity	756
Defined Atom Stereocenter Count	0
InChi Key	OAMPOKDQUZAAKE-UHFFFAOYSA-N
InChi Code	InChI=1S/C25H24N6O2S/c1-25(2,3)20-10-21(30-33-20)29-23(32)27-16-7-4-14(5-8-16)18-13-31-22-17-12-26-11-15(17)6-9-19(22)34-24(31)28-18/h4-5,7-8,10-13,26H,6,9H2,1-3H3,(H2,27,29,30,32)
Chemical Name	1-(5-tert-butyl-1,2-oxazol-3-yl)-3-[4-(10-thia-4,12,15-triazatetracyclo[7.6.0.02,6.011,15]pentadeca-1(9),2,5,11,13-pentaen-13-yl)phenyl]urea
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	Twelve blood cell lines were examined, and FLT3-IN-14 (Compound 9c) (0-10 μM; 24 hours) inhibited growth with an IC50 of 0.011-1.582 μM [1]. With a GI50 above 10 μM, FLT3-IN-14 (0-10 μM; 72 hours) shows minimal toxicity in resting lymphocytes [1]. Annexin-V-positive cells are accumulated by FLT3-IN-14 (1-50 nM; 24 and 48 hours) in a concentration- and time-dependent manner [1]. In both cell lines, FLT3-IN-14 (25–100 nM; 24 and 48 hours) significantly promotes G1 arrest [1]. FLT3 dephosphorylation is induced by FLT3-IN-14 (1-50 nM; 24 hours) [1].
ln Vivo	In a dose-dependent manner, FLT3-IN-14 (1.0 and 3.0 mg/kg; IP; once daily for 28 days) effectively inhibited tumor development without exhibiting any adverse effects [1].
Cell Assay	Cell proliferation assay Cell Types: MOLT-4, HL-60, KG-1, KG-1a, MOLM-13, MV4-11, NOMO-1, OCI-AML2, PL-21, THP-1, K-562, KCL-22[1] Tested Concentrations: 0-10 μM Incubation Duration: 24 hrs (hours) Experimental Results: Inhibited the proliferation of these 12 blood cell lines with IC50 of 0.011-1.582 μM. Cytotoxicity assay Cell Types: PBL[1] Tested Concentrations: 0-10 μM Incubation Duration: 72 hrs (hours) Experimental Results: demonstrated low toxicity in resting lymphocytes, GI50 greater than 10 μM. Apoptosis analysis Cell Types: MV4-11[1] Tested Concentrations: 1, 10 and 50 nM Incubation Duration: 24 and 48 hrs (hours) Experimental Results: Accumulation of Annexin-V positive cells in a concentration- and time-dependent manner. Cell cycle analysis Cell Types: MOLM-13 and MV-14[1] Tested Concentrations: 25, 50, 75 and 100 nM Incubation Duration: 24 and 48 hrs (hours) Experimental Results: Significant G1 arrest was induced in both cell lines. Western Blot Analysis Cell Types: MV-14[1] Tested Concentrations: 1, 10 and 50 nM Incubation Duration: 24 hrs (hours) Experimental Results: Induction of FLT3 dephosphorylation.
Animal Protocol	Animal/Disease Models: NOD/SCID female mice (subcutaneously (sc) (sc) implanted with MV4-11) [1] Doses: 1.0 and 3.0 mg/kg Route of Administration: IP; one time/day for 28 days Experimental Results: 1 mg/kg and 3 mg/ kg dose, tumor growth was Dramatically diminished by 44.1% and 55.2%, respectively.
References	[1]. Synthesis of 2H-Imidazo[2',1':2,3] [1,3]thiazolo[4,5-e]isoindol-8-yl-phenylureas with promising therapeutic features for the treatment of acute myeloid leukemia (AML) with FLT3/ITD mutations. Eur J Med Chem. 2022.

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	2.1161 mL	10.5807 mL	21.1613 mL
	5 mM	0.4232 mL	2.1161 mL	4.2323 mL
	10 mM	0.2116 mL	1.0581 mL	2.1161 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.