Physicochemical Properties
| Molecular Formula | C29H29FN4O3 |
| Molecular Weight | 500.56 |
| Appearance | Typically exists as solid at room temperature |
| 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 | PI3K/Akt/mTOR-IN-4 (compound 4r) (0-100 µM; 48 h) exhibited antiproliferative activity with IC50 values of 3.38, 5.03, 7.24, 21.08, and 23.96 µM for SiHa, HeLa, Ca Ski, LO2, and HEK-293t cells, respectively[1]. PI3K/Akt/mTOR-IN-4 (0-16 µM; 24 h) induced apoptosis and cell cycle arrest at the G2/M phase[1]. PI3K/Akt/mTOR-IN-4 (4, 8,16 µM; 24 h) reduced the expression of p-PI3K, p-Akt, p-mTOR, and β-tubulin in a dose-dependent manner[1]. |
| ln Vivo | PI3K/Akt/mTOR-IN-4 (0-400 μM; 0-96 h) is not toxic to zebrafish embryos[1]. |
| Cell Assay |
Western Blot Analysis[1] Cell Types: SiHa Tested Concentrations: 4, 8, 16 μM Incubation Duration: 24 h Experimental Results: Decreased the expression of phosphorylation of PI3K, Akt, and mTOR, β-tubulin in a dose-dependent manner. Cell Proliferation Assay[1] Cell Types: SiHa, HeLa, Ca Ski, LO2, HEK-293t Tested Concentrations: 0-100 µM Incubation Duration: 48 h Experimental Results: Inhibited cell proliferative with IC50s of 3.38, 5.03, 7.24, 21.08, 23.96 µM for SiHa, HeLa, Ca Ski, LO2, HEK-293t cells, respectively. Cell Cycle Analysis[1] Cell Types: SiHa Tested Concentrations: 0-16 µM Incubation Duration: 24 h Experimental Results: Induced cell cycle arrest at G2/M phase with G2/M phase cells accumulating from 5.24 % (Ctrl) to 28.37 % (16 μM). Apoptosis Analysis[1] Cell Types: SiHa Tested Concentrations: 0-16 µM Incubation Duration: 24 h Experimental Results: Induced apoptosis the percentage of apoptotic cells were increased from 11.62 % (Ctrl) to 98.56 % (16 μM). |
| Animal Protocol |
Animal/Disease Models:zebrafish embryos[1] Doses: 0, 12.5, 25, 50, 100, 200, 400 μM Route of Administration: 0-96 h Experimental Results: Showed no toxicity for zebrafish embryos. |
| References |
[1]. Design, synthesis, and biological evaluation of novel benzimidazole derivatives as anti-cervical cancer agents through PI3K/Akt/mTOR pathway and tubulin inhibition. Eur J Med Chem. 2024 Apr 16;271:116425. |
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 | 1.9978 mL | 9.9888 mL | 19.9776 mL | |
| 5 mM | 0.3996 mL | 1.9978 mL | 3.9955 mL | |
| 10 mM | 0.1998 mL | 0.9989 mL | 1.9978 mL |