Physicochemical Properties
| Molecular Formula | C27H21N5O6 |
| Molecular Weight | 511.49 |
| CAS # | 3031403-77-4 |
| 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 | Compound F16, or Topoisomerase I/II inhibitor 4, has the ability to intercalate into DNA and inhibit both Topo I and II activities as well as enzymatic activity in a dose-dependent manner, ranging from 0 to 100 μM[1]. Compound F16, also known as Topoisomerase I/II inhibitor 4, demonstrates strong anti-proliferative action with IC50 values of 20 and 50 nM, respectively, which is ten times lower than L02[1]. It also demonstrates a high expression level of Topo I and II enzymes in A375 and HCT116 cells. Compound F16 (topoisomerase I/II inhibitor 4) (0–40 nM, A375 cell) suppresses the development, invasion, and migration of cancer cells as well as triggering apoptosis in the cells[1]. |
| ln Vivo | In the A375 Xenograft Model, topoisomerase I/II inhibitor 4 (compound F16) (10–50 mg/kg; iv; every 2 days, for 7 days) inhibits tumor growth[1]. Compound F16, or topoisomerase I/II inhibitor 4, had a plasma clearance rate (CL) that was seven times lower than VP-16's (0.007 L/min/kg). The half-life of topoisomerase I/II inhibitor 4 in delayed elimination is 151 minutes[1]. |
| Cell Assay |
Cell Cycle Analysis[1] Cell Types: A375 cells Tested Concentrations: 20, 40 and 80 nM Incubation Duration: 24 hrs (hours) Experimental Results: The cell cycle blocked at the S phase in a dose-dependent manner. Apoptosis Analysis[1] Cell Types: A375 cells Tested Concentrations: 40, 80 and 120 nM Incubation Duration: 8, 16 and 24 hrs (hours) Experimental Results: Induced apoptosis rates for 8, 16 and 24 h were 7.14, 23.78 and 36.21%, respectively. Induced apoptosis rates for 40, 80 and 120 nM were 23.26, 36.21 and 55.94%, respectively. Western Blot Analysis[1] Cell Types: A375 cells Tested Concentrations: 40, 80 and 120 nM Incubation Duration: 24 hrs (hours) Experimental Results: The expression levels of Ccaspase-3, C-caspase-8, C-caspase-9, Bad and Bax increased in a dose-dependent manner, the Bcl-2 expression was obviously diminished. |
| Animal Protocol |
Animal/Disease Models: Female BALB/c nude mice (5−6 weeks)[1] Doses: 10 and 20 mg/kg Route of Administration: intravenous (iv) injection; Per 2 days, for 14 days. Experimental Results: Inhibited tumor growth in a melanoma xenograft mouse model and no apparent loss in body weight. Animal/Disease Models: Kunming male mice [1] Doses: 10 mg/kg Route of Administration: intravenous (iv) injection; for 5, 15, 30, 60, 120, 240 and 360 min. Experimental Results: 1.19 Parameter F16 VP-16 Dose (iv) mg/kg 10 10 Cmax (ng/mL) 26952 17712 Tmax (min) 5 5 AUCplasma (min*ng/mL) 2878363 409528 T1/2 (min) 151 45 Vd (L/Kg) 0.2341 0.432 CL (L/min/kg) 0.001 0.007 |
| References |
[1]. Optimization of the Natural Product Calothrixin A to Discover Novel Dual Topoisomerase I and II Inhibitors with Improved Anticancer Activity. J Med Chem. 2022 Jun 9;65(11):8040-8061. |
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 | 1.9551 mL | 9.7754 mL | 19.5507 mL | |
| 5 mM | 0.3910 mL | 1.9551 mL | 3.9101 mL | |
| 10 mM | 0.1955 mL | 0.9775 mL | 1.9551 mL |