Physicochemical Properties
| Molecular Formula | C24H24N2O2 |
| Molecular Weight | 372.459566116333 |
| Exact Mass | 372.183 |
| CAS # | 2513461-95-3 |
| PubChem CID | 163322279 |
| Appearance | Typically exists as solid at room temperature |
| LogP | 4.6 |
| Hydrogen Bond Donor Count | 0 |
| Hydrogen Bond Acceptor Count | 4 |
| Rotatable Bond Count | 4 |
| Heavy Atom Count | 28 |
| Complexity | 567 |
| Defined Atom Stereocenter Count | 0 |
| InChi Key | HGOZQJNNPSXGNT-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C24H24N2O2/c1-16-6-4-5-13-26(16)14-15-28-21-10-9-20-22-19(21)11-12-25-23(22)17-7-2-3-8-18(17)24(20)27/h2-3,7-12,16H,4-6,13-15H2,1H3 |
| Chemical Name | 12-[2-(2-methylpiperidin-1-yl)ethoxy]-16-azatetracyclo[7.7.1.02,7.013,17]heptadeca-1(16),2,4,6,9(17),10,12,14-octaen-8-one |
| 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 | Topoisomerase I inhibitor 5 (compound 14) exhibits modest cytotoxicity on normal cells and antiproliferative effect on cancer cell lines at concentrations of 0–50 μM within 48 hours [1]. Cell cycle arrest in the G1 phase of MCF-7 is induced by topoisomerase I inhibitor 5 (2-8 μM; 24 hours) [1]. The rate of apoptosis in MCF-7/ADR and MCF-7 cells is increased by topoisomerase I inhibitor 5 (2-8 μM; 48 hours) [1]. Topoisomerase I inhibitor 5 (1.5–6 μM; 24 hours) upregulates levels of pro-apoptotic proteins and downregulates those of anti-apoptotic proteins in MCF-7/ADR. It also boosts the production of cleaved-caspase-3 and cleaved-PARP in MCF-7 [1]. Topoisomerase I inhibitor 5 (0.1 μM; 24 hours) causes MCF-7/ADR cells to accumulate ROS, which in turn causes apoptosis [1]. In MCF-7/ADR cells, topoisomerase I inhibitor 5 (10 μg/ml; 24 hours) causes a rise in ADR and Rh123 accumulation [1]. In 24 hours, P-gp expression in MCF-7/ADR cells was decreased by 14.95% and 18.10% at 10 and 20 μM dosages of topoisomerase I inhibitor 5 (5, 10 and 20 μM) [1]. |
| ln Vivo | Tumor growth is markedly inhibited by topoisomerase I inhibitor 5 (1 mg/kg and 10 mg/kg; intravenously administered once every two days for 21 days) [1]. |
| Cell Assay |
Cell Proliferation Assay Cell Types: A549, HepG-2, MCF-7, MDA-MB-231, MCF-7/ADR and LO2 cells [1] Tested Concentrations: 0-50 μM Incubation Duration: 48 hrs (hours) Experimental Results: Demonstrated anti-proliferation The IC50 in active cancer cell lines, A549, HepG-2, MCF-7, MDA-MB-231, and MCF- were 2.39 ± 0.23 μM, 4.88 ± 0.29 μM, 1.32 ± 0.14 μM, 7.64 ± 0.35 μM, and 2.42 ± respectively. 0.14 μM is 7/ADR respectively; it has low cytotoxicity to LO2 cells, with an IC50 of 36.52 ± 2.36 μM. Cell cycle analysis Cell Types: MCF-7[1] Tested Concentrations: 2, 4 and 8 μM Incubation Duration: 24 hrs (hours) Experimental Results: Induced MCF-7 cell cycle arrest in G1 phase. Apoptosis analysis Cell Types: MCF-7 and MCF-7/ADR cells [1] Tested Concentrations: 2, 4 and 8 μM Incubation Duration: 48 hrs (hours) Experimental Results: Apoptosis was induced in MCF-7 cells in a dose-dependent manner and increased Apoptosis The cell apoptosis rate in MCF-7/ADR increased from 2.8% to 15.2%. Western Blot Analysis Cell Types: MCF-7[1] Tested Concentrations: 1.5, 3 and 6 μM in MCF-7; 5, 10 and 20 μM in MC |
| Animal Protocol |
Animal/Disease Models: Bal b/c nude mice (106 MCF-7 cells injected into the left flank for 7 days) [1] Doses: 1 mg/kg and 10 mg/kg Route of Administration: intravenous (iv) (iv)injection; once every two days , lasting for 21 days. Experimental Results: Dramatically diminished tumor growth, with a tumor inhibition rate of 32.4% at 1 mg/kg and 7.2% at 10 mg/kg. |
| References | [1]. Zhong H, Zhao M, Wu C, Zhang J, Chen L, Sun J. Development of oxoisoaporphine derivatives with topoisomerase I inhibition and reversal of multidrug resistance in breast cancer MCF-7/ADR cells. Eur J Med Chem. 2022;235:114300. |
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 | 2.6849 mL | 13.4243 mL | 26.8485 mL | |
| 5 mM | 0.5370 mL | 2.6849 mL | 5.3697 mL | |
| 10 mM | 0.2685 mL | 1.3424 mL | 2.6849 mL |