| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
FTO (fat mass and obesity-associated protein, a human demethylase) – IC50 = 4.5 μM against FTO demethylation [1]
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|---|---|
| ln Vitro |
Compound 7 inhibited FTO demethylation with an IC50 of 4.5 μM (tested at concentrations of 0.75, 1.5, 3, 6, 12.5, and 25 μM). [1]
Compound 7 showed potent antiproliferative activity against various cancer cell lines: A549 (human lung carcinoma) IC50 = 2.8 ± 0.1 μM; A549R (cisplatin-resistant A549) IC50 = 4.5 ± 0.3 μM; NB-4 (human leukemia) IC50 = 1.5 ± 0.1 μM; A261 (murine glioma) IC50 = 3.3 ± 0.02 μM; HLF (human lung fibroblasts) IC50 = 9.9 ± 0.1 μM. The cancer selectivity index (CSI = IC50 HLF/IC50 A549) was 3.5. [1] Compound 7 induced DNA damage in A549 cells, as evidenced by increased expression of γH2AX (a biomarker for DNA double-strand breaks) in a concentration-dependent manner. The tumor suppressor p53 also showed a concentration-dependent increase in expression, further confirming DNA double-strand damage. [1] Compound 7 induced autophagic cell death in A549 cells, as demonstrated by a significant increase in LC3-II protein expression and conversion from LC3-I to LC3-II after 24 h treatment. Downregulation of p62 confirmed protein degradation in autophagosomes. [1] In 3D multicellular A549 tumor spheroid assays, compound 7 (20 μM) caused a decrease in spheroid size over time, while untreated spheroids increased in size. [1] Compound 7 exhibited no effect on the expression of FTO protein in A549 cells. [1] |
| Enzyme Assay |
FTO demethylase inhibition assay: Compounds 1-10 were examined for their inhibitory activity on FTO demethylation. Compounds 1 and 7 were tested at concentrations of 0.75, 1.5, 3, 6, 12.5, and 25 μM. FB23 was used as a positive control. The inhibition curves were generated, and IC50 values were calculated. [1]
Molecular docking: The crystal structure of FTO (PDB code 3LFM) was used as the docking target. AutoDock was employed for docking runs. The binding interactions between FTO and compound 7 involved amino acid residues: His-231, Ser-229, Val-228, Tyr-108, Leu-109, Pro-93, Thr-92, Asp-233, Glu-234, Val-94, His-232, Arg-96, Ala-227, Trp-230, Tyr-106, and Leu-203. Among these, Val-228, Leu-109, Pro-93, Val-94, Ala-227, Trp-230, and Leu-203 are hydrophobic; the others are hydrophilic. Three hydrogen bonds were present between compound 7 and Ser-229 (2.7913, 2.9371, and 2.7378 Å). The stability of the FTO-7 complex is derived from hydrophobic and hydrogen bond interactions. [1] |
| Cell Assay |
Cytotoxicity assay (IC50 determination): Human lung carcinoma (A549), cisplatin-resistant A549 (A549R), human leukemia (NB-4), murine glioma (A261), and human lung fibroblasts (HLF) cells were treated with various concentrations of compounds for 72 h. Cell viability was assessed, and IC50 values were calculated. The maximum DMSO concentration was controlled at 1% (v/v). [1]
Western blotting for DNA damage markers: A549 cells were cultured with specified concentrations of compound 7 for 24 h. Protein expression of γH2AX and p53 was evaluated by Western blotting. GAPDH served as a loading control. [1] Western blotting for autophagy markers: A549 cells were treated with compound 7 for 24 h. LC3 protein levels were assessed by Western blotting. Tubulin served as a loading control. Conversion from LC3-I to LC3-II and downregulation of p62 were observed. [1] 3D multicellular tumor spheroid (MCTS) assay: A549 tumor spheroids were incubated with compound 7 (20 μM) at different times. Spheroid size changes were monitored and imaged. Untreated spheroids increased in size over time, while treated spheroids decreased in size. [1] FTO protein expression: A549 cells were treated with compound 7, and FTO protein expression was evaluated. No effect on FTO protein expression was observed. [1] |
| Toxicity/Toxicokinetics |
Cytotoxicity against normal human lung fibroblasts (HLF) showed an IC50 of 9.9 ± 0.1 μM, providing a cancer selectivity index of 3.5. [1]
|
| References | |
| Additional Infomation |
Compound 7 is a 1,8-naphthalimide derivative containing a tertiary nitrogen atom and a pyridine group. It was identified as a potent FTO demethylase inhibitor with promising anticancer activity. This is the first report of naphthalimide derivatives as FTO demethylase inhibitors. The compound induced DNA damage and autophagic cell death in A549 cells. The anticancer activity can be fine-tuned by modifying the 3-position of the naphthalimide ring and the side chain, with the order: -NO2 (compound 1) ≈ pyridine (compound 11) and tertiary nitrogen atom (compound 7) ≈ secondary nitrogen (compound 11) > primary nitrogen (compound 12). [1]
|
| Molecular Formula |
C22H20N4O3
|
|---|---|
| Molecular Weight |
388.42
|
| Exact Mass |
388.1535405
|
| Appearance |
Typically exists as solid at room temperature
|
| LogP |
2.3
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
29
|
| Complexity |
652
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CN(C)CCN1C(=O)C2=CC=CC3=CC(=CC(=C32)C1=O)NC(=O)C4=CC=CC=N4
|
| InChi Key |
KNCBJZSKAIQFIS-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C22H20N4O3/c1-25(2)10-11-26-21(28)16-7-5-6-14-12-15(13-17(19(14)16)22(26)29)24-20(27)18-8-3-4-9-23-18/h3-9,12-13H,10-11H2,1-2H3,(H,24,27)
|
| Chemical Name |
N-[2-[2-(dimethylamino)ethyl]-1,3-dioxobenzo[de]isoquinolin-5-yl]pyridine-2-carboxamide
|
| Synonyms |
FTO-IN-10; CHEMBL5397454; FTO Inhibitor 7;
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| 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)
|
| 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
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|---|---|
| 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
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] 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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5745 mL | 12.8727 mL | 25.7453 mL | |
| 5 mM | 0.5149 mL | 2.5745 mL | 5.1491 mL | |
| 10 mM | 0.2575 mL | 1.2873 mL | 2.5745 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.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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