Physicochemical Properties
| Molecular Formula | C22H23CLFN5O2 |
| Molecular Weight | 443.901726961136 |
| Exact Mass | 443.152 |
| CAS # | 2435562-99-3 |
| PubChem CID | 162670691 |
| Appearance | Typically exists as solid at room temperature |
| Hydrogen Bond Donor Count | 4 |
| Hydrogen Bond Acceptor Count | 7 |
| Rotatable Bond Count | 5 |
| Heavy Atom Count | 31 |
| Complexity | 668 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | C1CN(CCN1)CCO/N=C/2\C3=CC=CC=C3N=C2C4=C(NC5=C4C=C(C=C5)F)O.Cl |
| InChi Key | FYOZPHRPSXFBMH-PQAWYCKWSA-N |
| InChi Code | InChI=1S/C22H22FN5O2.ClH/c23-14-5-6-18-16(13-14)19(22(29)26-18)21-20(15-3-1-2-4-17(15)25-21)27-30-12-11-28-9-7-24-8-10-28;/h1-6,13,24,26,29H,7-12H2;1H/b27-20+; |
| Chemical Name | 5-fluoro-3-[(3E)-3-(2-piperazin-1-ylethoxyimino)indol-2-yl]-1H-indol-2-ol;hydrochloride |
| 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 | FLT3-IN-15 (Compound 36) (0-100 nM) has anti-proliferative action on MOLM14 cell line [1], with concentrations ranging from 0 to 100 nM. The FLT3-IN-15 (0-1 μM; 72 hours) demonstrates an excellent safety profile against different cancer cell lines and is more sensitive to MV4-11 cells than the K562 cell line [1]. In MV4-11 cells, FLT3-IN-15 (0.01-1 μM; 4 hours) potently inhibits the phosphorylation of Erk1/2 and STAT5 [1]. |
| ln Vivo | In all mice, FLT3-IN-15 (20 mg/kg; oral; once daily for 21 days) resulted in a swift and total tumor remission [1]. One female mouse died on day 6 from FLT3-IN-15 (2000 mg/kg; oral; single dose); the female's estimated LD50 was 4,950 mg/kg [1]. According to reference [1], FLT3-IN-15 (10 μM) inhibits hERG ligand binding by 21.4%. FLT3-IN-15 (10 mg/kg; oral and intravenous injection; single dose) exhibits a 42.6% significant increase in oral bioavailability, an AUClast of 25.0 μg·min/mL, and a Cmax of 36.5 ng/mL. FLT3-IN-15 pharmacokinetic parameters in male ICR mice [1]. PO (10 mg/kg) IV (10 mg/kg) AUClast (μg·min/mL) 25.0 ± 11.6 58.5 ± 57.4 AUCinf (μg·min/mL) 62.1 ± 58.6 103.4 ± 95.3 MRT (hr) 2811.3 ± 2713.0 1257.1 ± 1084.1 T1/2 (hour) 1775.7 ± 1901.0 1099.2 ± 98.7 VSS (L/kg) 127891 ± 104764 Cmax (ng/mL) 36.5 ± 24.3 Tmax (min) 390.0 ± 366.0 Xu, 24h (%) 0.001 ± 0.0 0.002 GI24h (%) 0.05 ± 0.24 ± 0.02 F (%) 42.9 |
| Cell Assay |
Cell Proliferation Assay Cell Types: MOLM14 wild-type cells, MOLM14-ITD cells, MOLM14-ITD-D835Y cells, MOLM14-ITD-F691L cells [1] Tested Concentrations: 0-100 nM Incubation Duration: Experimental Results: Demonstrated anti-proliferation against MOLM14 Active cell lines, GI50 in MOLM14 wild-type cells, MOLM14-ITD cells, MOLM14-ITD-D835Y cells and MOLM14-ITD-F691L cells are 4.88±0.67nM, 1.85±0.06nM, 1.87±0.36nM and 3.27±0.99nM ,respectively. Cell proliferation assay Cell Types: MV4-11, K562, A549, HepG2, MDA-MB-231, HCT-116, PC3 and SK-OV-3[1] Tested Concentrations: 0-1 μM Incubation Duration: 72 hrs (hours) Experimental Results: Displayed It is extremely sensitive to MV4-11 cells (GI50 = 1 nM) compared to the K562 cell line and exhibits a good safety profile against other cancer cell lines. Western Blot Analysis Cell Types: MV4-11[1] Tested Concentrations: 10 nM, 100 nM and 1 μM Incubation Duration: 4 hrs (hours) Experimental Results: Demonstrated strong blockade of STAT5 and Erk1/2 phosphorylation. |
| Animal Protocol |
Animal/Disease Models: BALB/c nu/nu (injection of MV4-11) [1] Doses: 20 mg / kg Route of Administration: PO; one time/day for 21 days Experimental Results: All mice experienced rapid and complete tumor response and no weight loss or any other signs of toxicity during dosing. Animal/Disease Models: Female ICR mice [1] Doses: 2000 mg/kg Route of Administration: Oral; Single Experimental Results:Caused death of one female mouse in the 2,000 mg/kg group on day 6, and determined the death of the male mouse The approximate lethal dose (ALD) exceeded 2,000 mg/kg and 2,000 mg/kg for female mice respectively; the LD50 value for female mice was calculated to be 4,950 mg/kg. Animal/Disease Models: Male ICR mouse[1] Doses: 10 mg/kg Route of Administration: oral and intravenous (iv) (iv)injection; single (pharmacokinetic/PK/PK analysis) Experimental Results: AUClast is 25.0 μg·min/mL, Cmax is 36.5 ng/mL , the oral bioavailability was Dramatically increased by 42.6%. |
| References |
[1]. Discovery of orally active indirubin-3'-oxime derivatives as potent type 1 FLT3 inhibitors for acute myeloid leukemia. Eur J Med Chem. 2020;195:112205. |
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 | 2.2528 mL | 11.2638 mL | 22.5276 mL | |
| 5 mM | 0.4506 mL | 2.2528 mL | 4.5055 mL | |
| 10 mM | 0.2253 mL | 1.1264 mL | 2.2528 mL |