Physicochemical Properties
| CAS # | 2755454-90-9 |
| 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 | Autophagy-IN-2 (Compound 7h) (0-200 μM, 48 hours) exhibits anti-viability efficacy against many cancer cell types [1]. In a p62-dependent manner, autophagy-IN-2 (0-20 μM, 0-48 h) downregulates chromatin ubiquitination, inhibiting autophagy flux and compromising DNA repair [1]. In 48 hours, autophagy-IN-2 (0–20 μM) causes both mitochondria-dependent intrinsic apoptosis and cell cycle arrest in S phase [1]. |
| ln Vivo | Autophagy-IN-2 (Compound 7h) (5 and 15 mg/kg; i.p.; every 3 days for 3 weeks) suppresses tumor development in a dose-dependent manner [1]. |
| Cell Assay |
Cell Viability Assay[1] Cell Types: MDA-MB-231, MDA-MB-468, SCC25, HSC-3, HCT116, SW620, PC3, Aspc, PNAC, U87, U251 and LN229 Tested Concentrations: 0-200 μM Incubation Duration: 48 hrs (hours) Experimental Results: demonstrated anti-vital activity, IC50 values were 8.31±1.05, 6.03±0.82, 18.64±2.92, 24.03±3.75, 35.53±7.52, 25.43±2.42, 17.48±1.27, 26.89±5.23, 19.25±3.96, against MDA-MB-231, MDA-MB-468, SCC25, HSC-3, HCT116, SW620, PC3, Aspc, PNAC, U87, U251 and LN229 cells were 15.37 ± 1.78, 12.92 ± 2.38 and 61.35 ± 11.61 μM, respectively. Western Blot Analysis[1] Cell Types: MDA-MB-231 and MDA-MB-468 Tested Concentrations: 5, 10 and 20 μM Incubation Duration: 0, 6, 12, 24, 36 and 48 hrs (hours) Experimental Results: Significant induction of LC3B - II causes p62 accumulation in TNBC cells in a dose- and time-dependent manner, increases γH2AX in a dose- and time-dependent manner, activates the phosphorylation of ATM, NBS1, and SMC1, increases p62 in the nucleus and cytoplasm in a dose-dependent manner, and Dramatically Dramatically reduces DNA damag |
| Animal Protocol |
Animal/Disease Models: Sixweeks old female NOD/SCID (severe combined immunodeficient) mouse, MDA-MB-231 xenograft [1] Doses: 5 mg/kg, 15 mg/kg Route of Administration: intraperitoneal (ip) injection, once every 3 days for 3 weeks Experimental Results: Inhibited human TNBC tumor growth in a dose-dependent manner and resulted in concentration-dependent upregulation of LC3B-II, p62, γH2AX and PARP. |
| References | [1]. Yang DL, et al. Discovery of fused benzimidazole-imidazole autophagic flux inhibitors for treatment of triple-negative breast cancer. Eur J Med Chem. 2022 Jun 26;240:114565. |
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.) |